Luminex 100 IS User Manual

Lum/nex

Luminex 100™ IS

User Manual Version 2.3

© LUMINEX CORPORATION, 2001-2005. All rights reserved. No part of this publication may be reproduced, transmitted, transcribed, or translated into any language or computer language, in any form or by any means without prior express, written consent of:

LUMINEX CORPORATION

12212 Technology Boulevard

Austin, Texas 78727-6115

U.S.A.

V oice: (512) 219-8020

Fax: (512) 219-5195

Luminex

Luminex Corporation (Luminex) reserves the right to modify its product s and services at any time. This guide is subject to change without notice. Although prepared to ensure accuracy, Luminex assumes no liability for errors or omissions, or for any damages resulting from the application or use of this information.

® 100™ IS User Manual Version 2.3

REF

CN-M018-01

PN 89-00002-00-072 Rev. C

October 2005

REP

Paul. A. Rowden 33 Stapleford Road Middlesbrough Cleveland TS39ES England (TQMUK@aol.com)

The following are trademarks of Luminex: Luminex, Luminex 100, Luminex 100 IS, LabMAP, xMAP, LumAvidin, Luminex XYP, Luminex FlexMAP, and Luminex SD. All other trademarks, including Windows,

Cheminert, Pentium, and Dell The Luminex 100 IS software uses the VideoSoft

ActiveX controls, which are copyrighted by VideoSof t, 2001. The contents of this manual and the associated Luminex software are the proper ty of L uminex and are

copyrighted. Except as specified in the End User License Agreement, any reproduction in whole or in part is strictly prohibited.

are trademarks of their respective companies.

® VsFlexGrid Pro 7.0, VsPrinter 7.0, and VsView 3.0

Standard Terms and Conditions For Use of Product

By opening the packaging containing this product ("Product") or by using such Product in any manner, you are consenting and agreeing to be bound by the following terms and conditions. You are also agreeing that the following terms and conditions constitute a legally valid and binding contract that is enforceable against you. If you do not agree to all of the terms and conditions set forth below, you must promptly return the Product for a full refund prior to using them in any manner.

1. Acceptance - ALL SALES ARE SUBJECT TO AND EXPRESSLY CONDITIONED UPON THE TERMS AND CONDITIONS CONTAINED HEREIN, AND UPON BUYER'S ASSENT THERETO. NO VARIATION OF THESE TERMS AND CONDITIONS SHALL BE BINDING UPON LUMINEX CORPORATION ("LUMINEX") UNLESS AGREED TO IN WRITING AND SIGNED BY AN AUTHORIZED REPRESENTATIVE OF LUMINEX. For purposes of this agreement, "Seller" shall mean the Luminex authorized reseller that sells the Product to Buyer. Buyer, by accepting the Product shall be deemed to have assented to the terms and conditions set forth herein, notwithstanding any terms contained in any prior or later communications from Buyer and whether or not Seller shall specifically or expressly object to any such terms.

2. Warranties - Any warranty obligations for the Product shall be exclusively provided in writing to Buyer directly by Seller. LUMINEX MAKES NO WARRANTY WHATSOEVER REGARDING THE PRODUCT AND LUMINEX SPEFICALLY DISCLAIMS ALL WARRANTIES, EXPRESS OR IMPLIED, INCLUDING ANY IMPLIED WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE. NEITHER SELLER NOR LUMINEX SHALL IN ANY EVENT BE LIABLE FOR INCIDENTAL, CONSEQUENTIAL OR SPECIAL DAMAGES OF ANY KIND RESULTING FROM ANY USE OR FAILURE OF THE PRODUCT, EVEN IF SELLER OR LUMINEX HAS BEEN ADVISED OF THE POSSIBILITY OF SUCH DAMAGE INCLUDING, WITHOUT LIMITATION, LIABILITY FOR LOSS OF WORK IN PROGRESS, DOWN TIME, LOSS OF REVENUE OR PROFITS, FAILURE TO REALIZE SAVINGS, LOSS OF PRODUCTS OF BUYER OR OTHER USE OR ANY LIABILITY OF BUYER TO A THIRD PARTY ON ACCOUNT OF SUCH LOSS, OR FOR ANY LABOR OR ANY OTHER EXPENSE, DAMAGE OR LOSS OCCASIONED BY SUCH PRODUCT INCLUDING PERSONAL INJURY OR PROPERTY DAMAGE UNLESS SUCH PERSONAL INJURY OR PROPERTY DAMAGE IS CAUSED BY SELLER'S GROSS NEGLIGENCE.

3. Buyer's Use of Product -Buyer agrees that no rights or licenses under Luminex's patents shall be implied from the sale of the Product, except as expressly provided herein, and Buyer does not receive any right under Luminex's patent rights hereunder. Buyer acknowledges and agrees that the Product is sold and licensed only for use with Luminex's standard fluorescently dyed microspheres. Buyer further acknowledges that the Product have not received approval from the United States Food and Drug Administration or other federal, state or local regulatory agencies and have not been tested by Seller or Luminex for safety or efficacy in food, drug, medical device, cosmetic, commercial or any other use, unless otherwise stated in Seller's technical specifications or material data sheets furnished to Buyer. Buyer expressly represents and warrants to Seller that Buyer will properly test and use any Product in accordance with the practices of a reasonable person who is an expert in the field and in strict compliance with the United States Food and Drug Administration and all applicable domestic and international laws and regulations, now and hereinafter enacted.

BUYER HEREBY GRANTS TO LUMINEX A NONEXCLUSIVE, WORLDWIDE, UNRESTRICTED, ROYALTY-FREE, FULLY PAID-UP LICENSE, WITH THE RIGHT TO GRANT AND AUTHORIZE SUBLICENSES, UNDER ANY AND ALL PATENT RIGHTS IN INVENTIONS COMPRISING MODIFICATIONS, EXTENSIONS, OR ENHANCEMENTS MADE BY BUYER TO THE PRODUCT OR TO THE MANUFACTURE OR USE OF THE PRODUCT ("IMPROVEMENT PATENTS"), TO MAKE, HAVE MADE, USE, IMPORT, OFFER FOR SALE OR SELL ANY AND ALL PRODUCT; EXPLOIT ANY AND ALL METHODS OR PROCESSES; AND OTHERWISE EXPLOIT IMPROVEMENT PATENTS FOR ALL PURPOSES. NOTWITHSTANDING THE FOREGOING, "IMPROVEMENT PATENTS" SPECIFICALLY EXCLUDES PATENT CLAIMS CONCEIVED AND REDUCED TO PRACTICE BY BUYER CONSISTING OF METHODS OF SAMPLE PREPARATION, METHODS OF CONJUGATING PRODUCT TO ANALYTES, THE COMPOSITION OF MATTER OF THE SPECIFIC CHEMISTRIES OF THE ASSAYS DEVELOPED BY BUYER AND METHODS OF PERFORMING THE ASSAYS (I.E., THE PROTOCOL FOR THE ASSAY). Buyer has the responsibility and hereby expressly assumes the risk to verify the hazards and to conduct any further research necessary to learn the hazards involved in using the Product. Buyer also has the duty to warn Buyer's customers, employees, agents, assigns, officers, successors and any auxiliary or third party personnel (such as freight handlers, etc.) of any and all risks involved in using or handling the Product. Buyer agrees to comply with instructions, if any, furnished by Seller or Luminex relating to the use of the Product and not misuse the Product in any manner. Buyer shall not reverse engineer, decompile, disassemble or modify the Product. Buyer acknowledges that Luminex retains ownership of all patents, trademarks, trade secrets and other proprietary rights relating to or residing in the Product.

4. Buyer's Representations, Release and Indemnity - Buyer represents and warrants that it shall use the Product in accordance with Paragraph 2, "Buyer's Use of Product," and that any such use of Product will not violate any law, regulation, judicial order or injunction. Buyer agrees to release, discharge, disclaim and renounce any and all claims, demands, actions, causes of action and/or suits in law or equity, now existing or hereafter arising, whether known or unknown, against Seller and Luminex, and their respective officers, directors, employees, agents, successors and assigns (collectively the "Released Parties"), with respect to the use of the Product. Buyer agrees to indemnify and hold harmless the Released Parties from and against any suits, losses, claims, demands, liabilities, costs and expenses (including attorney, accounting, expert witness, and consulting fees) that any of the Released Parties may sustain or incur as a result of any claim against such Released Party based upon negligence, breach of warranty, strict liability in tort, contract or any other theory of law or equity arising out of, directly or indirectly, the use of the Product or by reason of Buyer's failure to perform its obligations contained herein. Buyer shall fully cooperate with the Released Parties in the investigation and determination of the cause of any accident involving the Product which results in personal injury or property damage and shall make available to the Released Parties all statements, reports, recordings and tests made by Buyer or made available to Buyer by others.

5. Patent Disclaimer - Neither Seller nor Luminex warrants that the use or sale of the Product will not infringe the claims of any United States or other patents covering the product itself or the use thereof in combination with other products or in the operation of any process.

End-User License Agreement (EULA) for Luminex® Software

This Luminex End-User License Agreement (“EULA”) is a legal agreement between you (either an individual or a single entity , also referred herein as “you”) the end-user and Luminex Corporation (“Luminex”) regarding the use of the Luminex software product identified above, which includes computer software and online or electronic documentation and may include associated media and printed materials (if any) (“SOFTWARE PRODUCT” or “SOFTWARE”).

The SOFTWARE PRODUCT is protected by copyright laws and international copyright treaties, as well as other intellectual property laws and treaties. The SOFTWARE PRODUCT is licensed, not sold.

1. GRANT OF LICENSE. Subject to the terms and conditions of this EULA, Luminex hereby grants to you a nonexclusive, nontransferable, nonassignable license (without right to sublicense) under Luminex’s copyrights and trade secrets to use the SOFTW ARE PRODUCT on a hardware platform purchased from Luminex pursuant to Luminex’s terms and conditions of sale. You may make one (1) copy of the SOFTWARE PRODUCT for backup or archival purposes only. Although no rights or licenses under any of Luminex's patents are granted by or shall be implied from the license of the SOFTW ARE or the sal e of Luminex instrumentation to you, the purchaser, you may obtain a license under Luminex’ s patents, if any, to use this unit of Luminex instrumentation with fluorescently labeled microsphere beads authorized by Luminex by purchasing such beads from Luminex or an authorized Luminex reseller.

2. RESTRICTIONS.

• You must maintain all proprietary notices on all copies of the SOFTWARE PRODUCT.

• You may not distribute copies of the SOFTWARE PRODUCT to third parties.

• You may not reverse-engineer, decompile, disassemble, or otherwise attempt to derive source code from the SOFTWARE PRODUCT.

• You may not copy (other than one backup or archival copy), distribute, sublicense, rent, lease, transfer or grant any rights in or to all or any portion of the SOFTWARE PRODUCT.

• You must comply with all applicable laws regarding the use of the SOFTWARE PRODUCT.

• You may not modify or prepare derivative works of the SOFTWARE PRODUCT.

• You may not use the SOFTWARE PRODUCT in a computer-based service business or publicly display visual output of the SOFTWARE PRODUCT.

• You may not transmit the SOFTWARE PRODUCT over a network, by telephone, or electronically by any means.

3. TERM AND TERMINA TION. Your rights under this EULA are effective until termination. You may terminate this EULA at any time by destroying the SOFTWARE PRODUCT, including all computer programs and documentation, and erasing any copies residing on your computer equipment. Luminex may terminate this EULA upon thirty (30) days written notice to you. Your rights under this EULA automatically terminate without further action on the part of Luminex if you do not comply with any of the terms or conditions of this EULA. Upon any termination of this EULA, you agree to destroy the SOFTWARE PRODUCT and erase any copies residing on your computer equipment.

4. RIGHTS IN SOFTWARE. All rights and title in and to the SOFTWARE PRODUCT and any copies thereof are owned by Luminex or its suppliers. This EULA is not a sale and does not transfer to you any title or ownership interest in or to the SOFTWARE or any p atent, copyright, trade secret, trade n ame, trademark or other intellectual property right therein. You shall not remove, alter, or obscure any proprietary notices contained on or within the SOFTWARE and shall reproduce such notices on any back-up copy of the SOFTWARE. All title and intellectual property rights in and to the content which may be accessed through use of the SOFTWARE PRODUCT is the property of the respective content owner and may be protected by applicable copyright or other intellectual property laws and treaties. This EULA grants you no rights to use such content.

5. EXPORT RESTRICTIONS. You agree that you will not export or re-export the SOFTWARE PRODUCT to any country , person, entity, or end-user subject to U.S.A. export restrictions. Y ou hereby warrant no state or federal agency has suspended, revoked, or denied your export privileges.

6. NO WARRANTY. THE SOFTWARE PRODUCT IS LICENSED “AS IS.” ANY USE OF THE SOFTW ARE PRODUCT IS A T YOUR OWN RISK. THE SOFTWARE PRODUCT IS PROVIDED FOR USE ONLY WITH LUMINEX PRODUCTS. TO THE MAXIMUM EXTENT PERMITTED BY APPLICABLE LAW, LUMINEX AND ITS SUPPLIERS DISCLAIM ALL WARRANTIES, EITHER EXPRESS OR IMPLIED, INCLUDING , BUT NOT LIMITED TO, IMPLIED W ARRANTIES OF MERCHANT ABILITY, FITNESS FOR A PARTICULAR PURPOSE, AND NONINFRINGEMENT.

7. LIMITATION OF LIABILITY. IN NO EVENT SHALL LUMINEX OR ITS SUPPLIERS BE LIABLE FOR ANY SPECIAL, INCIDENTAL, INDIRECT, OR CONSEQUENTIAL DAMAGES WHATSOEVER (INCLUDING, WITHOUT LIMITATION, DAMAGES FOR LOSS OF BUSINESS PROFITS, BUSINESS INTERRUPTION, LOSS OF BUSINESS INFORMA TION, OR ANY OTHER PECUNIARY LOSS) ARISING OUT OF THE USE OF OR INABILITY TO USE THE SOFTWARE PRODUCT, EVEN IF LUMINEX HAS BEEN ADVISED OF THE POSSIBILITY OF SUCH DAMAGES.

MISCELLANEOUS. This EULA is governed by the laws of the State of Texas, U.S.A., without reference to conflicts of laws principles. You shall not assign or sublicense or otherwise transfer the rights or license granted hereunder, by agreement or by operation of law, without the prior written consent of Luminex, and all assignments in violation of this prohibition shall be null and void. This EULA is the complete and exclusive agreement of Luminex and you and supersedes all other communications, oral or written, relating to the subject matter hereof. No change to this EULA shall be valid unless in writing and signed by the party against whom enforcement is sought. The waiver or failure of Luminex or you to exercise in any respect any right or rights provided for herein shall not be deemed a waiver of any further right hereunder. If any provision of this EULA is held unenforceable, the remainder of this EULA will continue in full force and effect.

EULA PN: 89-30000-00-070

Introduction 1-1

About This Manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1-1

The Luminex 100 IS 2.3 System. . . . . . . . . . . . . . . . . . . . . . . . . . .1-1

Intended Use. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1-1

Technical Support . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1-2

Luminex Website . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1-2

Safety 2-1

Symbols. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2-1

Warnings and Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2-2

Safety Precautions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-2

FCC Label. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2-4

Fluidics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2-4

Luminex 100 Analyzer Laser . . . . . . . . . . . . . . . . . . . . . . . . . . 2-5

Barcode Reader Laser. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2-7

Mechanical . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2-7

Biological . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2-7

Heat. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-8

Blue Indicator Light . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2-8

Decontaminating the Luminex 100 Analyzer

for Return Shipment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-9

The System 3-1

Theory of Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-1

Hardware. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3-2

xMAP Technology Reagents. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-3

Required Laboratory Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . .3-3

Luminex 100 IS 2.3 Software . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-3

Luminex 100 IS Performance Specification . . . . . . . . . . . . . . . . . .3-3

Speed. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-3

Accuracy and Precision . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-4

Sensitivity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-4

Capacity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3-4

Luminex 100 Analyzer General . . . . . . . . . . . . . . . . . . . . . . . . . . .3-5

Optics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3-5

Fluidics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3-5

PN 89-00002-00-072 Rev. C i

Luminex 100 IS User Manual Version 2.3 xMAP Technology

Electronics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-5

Luminex XYP Instrument General. . . . . . . . . . . . . . . . . . . . . . . . . 3-6

Luminex SD System General. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-6

PC Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-6

Recommended Additional Equipment. . . . . . . . . . . . . . . . . . . 3-7

Uninterruptible Power Supply (UPS) . . . . . . . . . . . . . . . . 3-7

Surge Protector . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-7

Printer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-7

Barcode Labels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-7

Vortex . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-7

Bath Sonicator. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-7

System Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-7

Electronics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-7

Power Input Module . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-7

Communications Ports(SB9-PIN). . . . . . . . . . . . . . . . . . . 3-7

Luminex 100 Analyzer Ventilation Filter. . . . . . . . . . . . . 3-7

Luminex XYP Instrument Ventilation Filter. . . . . . . . . . . 3-8

Fluidics. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-8

Sample Arm. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-8

Luminex XYP Instrument Sample Probe. . . . . . . . . . . . . 3-8

Cheminert® Fitting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-8

Access Doors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-9

Air Intake Filter. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-10

Syringe. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-10

Sheath Filter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-11

Air, Waste, and Sheath Fluid Connectors . . . . . . . . . . . . 3-11

Luminex Sheath Delivery System . . . . . . . . . . . . . . . . . 3-12

Waste Fluid Container. . . . . . . . . . . . . . . . . . . . . . . . . . . 3-12

Optical . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-12

xMAP Technology Reagents. . . . . . . . . . . . . . . . . . . . . . . . . 3-12

Basic Concepts 4-1

Background Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-1

Fluidics. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-1

Excitation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-1

xMAP Microspheres. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-2

Software Overview. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-2

Using Luminex 100 IS 2.3 Software 5-1

Main Window. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-1

Menu Bar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-3

Toolbar. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-4

Tabs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-4

Home tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-4

Run Batch Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-5

Maintenance Tab. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-8

Diagnostics tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-9

Acquisition Detail Tab . . . . . . . . . . . . . . . . . . . . . . . . . . 5-12

ii PN 89-00002-00-072 Rev. C

xMAP Technology Contents

Status Bar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-17

Secondary Windows . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-20

Batch Setup Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-20

Analysis Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-22

Errors Tab. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-23

Standards Tab. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-24

Samples Tab. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-28

Replicate Averaging. . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-29

Commands. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-31

Acquire Patient . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-31

Add Batch. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-31

Alcohol Flush . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-31

Autoscale . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-31

Autosize . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-31

Backflush . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-31

CAL1 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-31

CAL2 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-31

Cancel (Command). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-31

Cancel All. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-31

Change Lot . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-31

CON1 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-31

CON2 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-31

Connect to Instrument. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-32

Create New Multi-Batch. . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-32

Delete Batch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-32

Density/Decaying . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-32

Display Confirmation Screens . . . . . . . . . . . . . . . . . . . . . . . .5-32

Disconnect from the Instrument . . . . . . . . . . . . . . . . . . . . . . . 5-32

Drain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-32

Eject/Retract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-32

Enable Raw Data Storage. . . . . . . . . . . . . . . . . . . . . . . . . . . .5-32

Export Batch Data. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-32

Export CAL. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-32

Export CON . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-32

Export Data. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-33

Help. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-33

Import Calibration. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-33

Import Control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-33

Import Template . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-33

Insert . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-33

Invalidate Control. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-33

Invalidate Standard. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-33

Load Patient List. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-33

Log/Linear . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-33

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Maximize/Minimize. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-33

Next Test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-33

New Advanced Batch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-34

New Batch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-34

New CAL Targ. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-34

New CON Targ.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-34

New Lot . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-34

Open Batch. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-34

Open Help . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-34

Open Incomplete Batch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-34

Open Multi-Batch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-34

Pause . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-34

Previous Test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-35

Prime . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-35

Print Batch Worklist . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-35

Print Report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-35

Recalc. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-35

Replay Batch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-35

Report Raw Fluorescence . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-35

Resume. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-35

Sample Probe Down . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-35

Sanitize. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-35

Save and Load . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-36

Save Only . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-36

Self Diagnostics. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-36

Show Bead. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-36

Single Step. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-36

Skip [wells] . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-36

Soak . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-36

Start Analysis. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-37

Start (Plate) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-37

Statistics. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-37

Test Sort Orders. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-37

Validate Control. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-37

Validate Standard . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-37

View Batch Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-37

Warmup . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-37

Wash . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-37

Zoom . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-37

Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-38

Using the Online Help . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-38

Setting Software Options. . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-39

Define General Tab Information. . . . . . . . . . . . . . . . . . . 5-39

Define Company Information Tab . . . . . . . . . . . . . . . . . 5-40

iv PN 89-00002-00-072 Rev. C

xMAP Technology Contents

Define Data Export Tab Information. . . . . . . . . . . . . . . .5-41

Setting up the Favorites List. . . . . . . . . . . . . . . . . . . . . . . . . .5-42

Startup Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-43

Warm Up the System . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-43

Prime the System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-43

Backflush the System. . . . . . . . . . . . . . . . . . . . . . . . . . . .5-44

Run an Alcohol Flush. . . . . . . . . . . . . . . . . . . . . . . . . . . .5-44

Run a Wash Cycle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-44

Set Luminex XYP Instrument Heater Temperature. . . . .5-45

Calibration Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-46

Run System xMAP Calibrators . . . . . . . . . . . . . . . . . . . .5-47

Run System xMAP Controls . . . . . . . . . . . . . . . . . . . . . .5-49

Selecting Existing CAL or CON Lots . . . . . . . . . . . . . . .5-50

Importing CAL or CON Lots. . . . . . . . . . . . . . . . . . . . . .5-50

Exporting CAL or CON lots . . . . . . . . . . . . . . . . . . . . . . 5-51

Batch Setup Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-51

Importing Templates . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-52

Create a New Batch . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-53

Open a Batch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-55

Copying and Exporting Batch Data . . . . . . . . . . . . . . . . .5-55

Clear a Batch from the System. . . . . . . . . . . . . . . . . . . . .5 -55

Replay a Batch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-56

Delete a Batch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-58

Create a Multi-Batch . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-58

Open a Multi-Batch . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-60

Process Multiple Plates . . . . . . . . . . . . . . . . . . . . . . . . . .5-60

Re-Run or Recover Incomplete Batch . . . . . . . . . . . . . . .5-61

Scan in Samples With a Barcode Reader. . . . . . . . . . . . .5-62

Add a Patient List. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-62

Edit a Patient List. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-64

Assign Sample Dilution Factors. . . . . . . . . . . . . . . . . . . .5-65

Create a New Advanced Batch. . . . . . . . . . . . . . . . . . . . .5-66

Managing Assay Lots . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-71

Create a New Lot . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-72

Edit Lot Information on an Unused Template . . . . . . . . .5 -73

Edit Lot Information on a Used Template . . . . . . . . . . . .5-74

Import Lot to an Existing Template. . . . . . . . . . . . . . . . .5-75

Export a Lot from an Existing Template . . . . . . . . . . . . .5-76

Analyzing Batches and Multi-Batches . . . . . . . . . . . . . . . . . .5-76

Customize Data Analysis Settings . . . . . . . . . . . . . . . . . .5-77

Enable Automatic Analysis . . . . . . . . . . . . . . . . . . . . . . .5-82

Analyze Processed Batch Data. . . . . . . . . . . . . . . . . . . . .5-83

View Detailed Test Analysis . . . . . . . . . . . . . . . . . . . . . .5-84

Validating or Invalidating Standards and Controls . . . . . 5-84

Change Lot. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-85

Running Reports and Analyses. . . . . . . . . . . . . . . . . . . . . . . .5-87

Report Types . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-87

Export Batch Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-89

Print Data Analysis Report. . . . . . . . . . . . . . . . . . . . . . . .5 -90

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Database Management Procedures . . . . . . . . . . . . . . . . . . . . 5-91

Back Up the Database . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-91

Delete Database Entries . . . . . . . . . . . . . . . . . . . . . . . . . 5-92

Restore Database Data. . . . . . . . . . . . . . . . . . . . . . . . . . . 5-92

Maintenance Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-93

Drain the Analyzer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-94

Run Self-Diagnostics. . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-95

Using the Cleanup Utility . . . . . . . . . . . . . . . . . . . . . . . . 5-95

Daily Shutdown Procedures. . . . . . . . . . . . . . . . . . . . . . . . . . 5-97

Sanitize the System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-97

Run a Wash Command . . . . . . . . . . . . . . . . . . . . . . . . . . 5-97

Perform a Soak Command . . . . . . . . . . . . . . . . . . . . . . . 5-98

Exit Luminex IS 2.3 Software. . . . . . . . . . . . . . . . . . . . . 5-98

Maintenance and Cleaning 6-1

Daily Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-1

Before Running Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-1

After Running Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-2

Routine Tasks. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-2

Sheath and Waste Fluids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-2

Refill the Sheath Fluid Container . . . . . . . . . . . . . . . . . . . 6-3

Empty the Waste Container. . . . . . . . . . . . . . . . . . . . . . . . 6-3

Weekly . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-3

Visual Inspection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-3

Run Self-Diagnostics. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-3

Clean Sample Probe. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-3

Flush the System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-4

Monthly . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-4

Clean the Sample Probe. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-4

Clean Exterior Surfaces . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-4

Calibration and System Controls. . . . . . . . . . . . . . . . . . . . . . . 6-5

Every Six Months. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-5

Luminex 100 Analyzer Air Intake Filter . . . . . . . . . . . . . . . . . 6-5

Luminex XYP Instrument Air Intake Filter. . . . . . . . . . . . . . . 6-6

Syringe Seal. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-8

Luminex 100 Analyzer Ventilation Filter . . . . . . . . . . . . . . . . 6-9

Annually. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-10

Sheath Filter. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-10

As required . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-11

Fuses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-11

Maintenance Log . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-13

Troubleshooting 7-1

Troubleshooting the Luminex 100 IS System . . . . . . . . . . . . . . . . 7-1

Power Supply Problems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-2

Communication . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-2

vi PN 89-00002-00-072 Rev. C

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Pressurization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-3

Fluid Leaks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7-4

Sample Probe. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-5

Calibration and Control Problems. . . . . . . . . . . . . . . . . . . . . . . . . .7-7

Acquisition Problems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7-9

Bead Detail Irregularities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-10

Error States . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7-13

System Error Messages. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7-13

Sample Error Messages. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7-15

Luminex SD Problems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7-18

Filter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7-18

Malfunction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-18

Draining the Reservoir . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-18

Verification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-19

Product Numbers 8-1

Hardware. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .8-1

Software. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .8-3

xMAP Reagents. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-3

Training. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-3

Glossary A-1

Luminex 100 IS System Installation B-1

Overview. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B-1

Luminex 100 IS System Setup . . . . . . . . . . . . . . . . . . . . . . . . . . . B-1

Connect the Luminex 100 analyzer and

Luminex XYP to the PC. . . . . . . . . . . . . . . . . . . . . . . . . . . . . B-3

Install the Luminex XYP Instrument Sample Probe . . . . . . . B-6

Power On System Components . . . . . . . . . . . . . . . . . . . . . . . B-8

Accept the Luminex 100 IS 2.3 Software

License Agreement. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B-8

Adjust the Sample Probe Vertical Height. . . . . . . . . . . . . . . . B-8

Install the Luminex XYP Instrument Reservoir. . . . . . . . . . . B-9

Calibrate and Verify the System. . . . . . . . . . . . . . . . . . . . . . B-10

Install the SD System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B-10

Install the Luminex XYP Instrument Heater Block . . . . . . . B-13

Luminex 100 IS 2.3 Software Installation. . . . . . . . . . . . . . . . . . B-14

Luminex 100 Version 1.7 with Windows 98 to

Luminex 100 IS Version 2.3. . . . . . . . . . . . . . . . . . . . . . . . . B-15

Install New PC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B-15

Install Luminex 100 IS 2.3 Software . . . . . . . . . . . . . . . B-15

Verify Successful Upgrade. . . . . . . . . . . . . . . . . . . . . . . B-16

Luminex 100 Version 1.7 with Windows 2000 to

Luminex 100 IS Version 2.3. . . . . . . . . . . . . . . . . . . . . . . . . B-17

Archive “My Sessions” folder . . . . . . . . . . . . . . . . . . . . B-17

PN 89-00002-00-072 Rev. C vii

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Remove Luminex LMAT Software . . . . . . . . . . . . . . . .B-17

Remove Luminex 100 Version 1.7 Software . . . . . . . . . B-18

Install Luminex 100 IS 2.3 Software. . . . . . . . . . . . . . . .B-19

Luminex 100 IS Version 2.1/2.2 to Luminex 100 IS

Version 2.3. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B-19

Backup Luminex 100 IS 2.1 or 2.2 Database . . . . . . . . . B-19

Remove Luminex 100 IS 2.1 or 2.2 . . . . . . . . . . . . . . . . B-20

Install Luminex 100 IS 2.3 Software. . . . . . . . . . . . . . . .B-20

Luminex 100 IS 2.3 Firmware Installation. . . . . . . . . . . . . . . . . .B-21

Firmware Upgrade Cable Configurations . . . . . . . . . . . . . . . B-21

Luminex 100 Analyzer Firmware Upgrade. . . . . . . . . . . . . . B-22

Connect Cable . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B-22

Upgrade Firmware. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .B-22

Verify Successful Firmware Upgrade . . . . . . . . . . . . . . B-23

Update Interface Cable . . . . . . . . . . . . . . . . . . . . . . . . . .B-24

Luminex XYP Instrument Firmware Update. . . . . . . . . . . . . B-24

Update Firmware. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .B-24

Verify Successful Firmware Upgrade. . . . . . . . . . . . . . . B-24

Luminex SD System Firmware Update. . . . . . . . . . . . . . . . .B-25

Update Firmware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B-25

Verify Successful Firmware Upgrade. . . . . . . . . . . . . . . B-26

Network Installation Advisory. . . . . . . . . . . . . . . . . . . . . . . . . . . B-26

Prepare System for First Use . . . . . . . . . . . . . . . . . . . . . . . . . . . .B-26

Installation Drawing. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B-27

Output.CSV C-1

Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C -1

Overall Design . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .C-1

Blank Lines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-2

Field Definitions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-2

Statistics Definitions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .C-4

Statistics Column Definitions . . . . . . . . . . . . . . . . . . . . . . . . . C-5

Luminex 100 IS OUTPUT.CSV file with no

additional features enabled . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-7

Luminex 100 IS OUTPUT.CSV file with all

additional features enabled. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-8

Index Index-1

viii PN 89-00002-00-072 Rev. C

Introduction

About This Manual This manual provides you with information to understand and use the

Luminex appendices that take you from this introduction to complete system operation.

The manual’s text and figures offer examples when necessary. Procedures are presented as step-by-step instructions. Glossary and index sections assist as references.

The conventions in this manual assume that the reader has a basic familiarity with computers, xMAP Microsoft common methods of accessing a command, such as from the main menu bar, from the toolbar, and from menus that appear when you right-click an area of the screen. Refer to the Glossary appendix for unfamiliar terminology.

® 100™ IS system. It consists of multiple chapters and

® technology, and a knowledge of

® Windows® software. We typically document the

The Luminex 100 IS

2.3 System

Intended Use The Luminex 100 IS 2.3 system is designed to perform a wide range

PN 89-00002-00-072 Rev. C 1 - 1

The Luminex 100 IS 2.3 system is a benchtop system consisting of the Luminex 100 analyzer, computer, monitor, keyboard, mouse, Luminex XY Platform instrument (Luminex XYP™), Luminex Sheath Delivery System (Luminex SD™), software, barcode reader, sheath and waste containers, and xMAP technology reagents.

of xMAP technology-based laboratory tests, measuring biomolecular reactions on the surface of xMAP microspheres. The system is intended For In-V itro Di agnostics Use for Export Only.

Luminex 100 IS User Manual Version 2.3 xMAP Technology

Technical Support Luminex Technical Support representatives are ready to help you,

particularly when the system or software cause any questions or problems. If the question or problem relates to materials from the assay kit, you should contact the kit provider directly.

Luminex Technical Support is available to users in the U.S. and Canada by calling 1-877-785-BEAD (-2323) between the hours of 7:00 a.m. and 7:00 p.m. Central Time, Monday through Friday. Users outside of the U.S., Canada, Europe can contact us at +1 512381-4397 between the hours of 7:00 a.m. to 7:00 p.m. Central Time, Monday through Friday. Inquiries may also be sent by email to support@luminexcorp.com.

Luminex Technical Support is also a vaila ble to users in Europe by calling +31-162408333 between the hours of 8:30 and 5:30, Central European Time, Monday through Friday. Email inquiries in Europe can be sent to supporteurope@luminexcorp.com.

Luminex Website Additional information is available on the Luminex website. Search

on the desired topic or navigate through menus. Also, review the website’s FAQ section.

 To access Luminex website FAQ section:

In your browser’s address field, enter: http://luminexcorp.custhelp.com. This address takes you directly to the FAQ section.

1 - 2 PN 89-00002-00-072 Rev. C

Safety

Symbols Please become familiar with the information in this chapter before

using the equipment. Do not perform procedures on your Luminex 100 IS 2.3 system that are not specifically contained in this manual, unless you are directed to do so by Luminex Technical Support.

These symbols describe warnings, cautions, and general information used in the operation of this instrument. These symbols are further defined under “Safety Precautions.”

Number Symbol Description Number Symbol Description

1 Alternating current (ac) 7 Warning (refer to manual)

2 Protective ground 8 Warning (refer to manual)

3 On 9 Warning (refer to manual)

4 Off 10 Warning (refer to manual)

5 SN Serial number 11 Warning (refer to manual)

6 Warning (refer to manual)

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Warnings and Notes Informational notes and warnings appear in this manual.

Note: A note provides general helpful information. No safety or performance issues are involved.

Caution: This message is used in cases where the hazard is minor or only potential hazard is present. Failure to comply with the caution may result in potentially hazardous conditions.

Warning: This message is used in cases where danger to the operator or to the performance of the instrument is present. Failure to comply with the warning may result in incorrect performance, instrument failure, invalid results, or hazard to the operator.

Danger: This message is used in cases where significant risk of serious injury or death is present.

Safety Precautions Read the following safety information before setting up or using the

Luminex 100 IS 2.3 system. A user should be present during operation. This system contains electrical, mechanical, and laser components which, if handled improperly, are potentially harmful. In addition, biological hazards may be present during system operation. Therefore, we recommend that all system users become familiar with the specific safety advisories below, in addition to adhering to standard laboratory safety practices. The protection provided by the equipment may be impaired or the warranty voided if the system is used in a manner not specified by the instructions or by Luminex Corporation.

This caution label appears on the back of the Luminex 100 analyzer and on the Luminex XYP instrument.

Figure 2-1 Fuse Caution Label

2 - 2 PN 89-00002-00-072 Rev. C

xMAP Technology Safety

Do not perform any maintenance or cleaning of the system’s electrical components, with the exception of replacing fuses.

This label appears on the back panel of the Luminex 100 analyzer and on the back panel of the Luminex XYP instrument.

Figure 2-2 CE Label

The Luminex 100 analyzer and the Luminex XYP instrument comply with European Union (EU) safety requirements and, therefore, may be marketed in the Europe Single Market.

This voltage label appears on the back of the Luminex 100 analyzer:

Figure 2-3 Luminex 100 Serial Number Label

The Luminex 100 analyzer has been tested by Underwriter Laboratories, Inc.® (UL).

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Luminex 100 IS User Manual Version 2.3 xMAP Technology

The following label appears on the back of the Luminex XYP instrument.

Figure 2-4 Luminex XYP Serial Number Label

The Luminex XYP instrument has been tested by UL.

FCC Label This equipment has been tested and found to comply with the limits

for a Class B digital device, pursuant to part 15 of the FCC Rules. These limits are designed to provide reasonable protection against harmful interference in a residential installation. This equipment generates, uses and can radiate radio frequency energy and, if not installed and used in accordance with the instructions, may cause harmful interference to radio communications. However, there is no guarantee that interference will not occur in a particular installation. If this equipment does cause harmful interference to radio or television reception, which can be determined by turning the equipment off and on, the user is encouraged to try to correct the interference by one or more of the following measures:

• Reorient or relocate the receiving antenna.

• Increase the separation between the equipment and the receiver.

• Connect the equipment into an outlet on a circuit different from that to which the receiver is connected.

• Consult the dealer or an experienced radio/TV technician for help.

Fluidics This system contains fluidics. In the event of a fluid leak, turn off all

power to the system and disconnect all power cords. Remember that the on/off switch is not a disconnect means; the power cord must be removed from the outlet. Contact Luminex Corporation for further information.

2 - 4 PN 89-00002-00-072 Rev. C

xMAP Technology Safety

You must monitor waste levels manually. Do not allow the waste container to overflow! Empty the waste container each time the sheath fluid container is filled. Do not place the waste container on top of the instrument.

Warning: If biological samples have been tested with the system,

use your standard laboratory safety practices when handling system waste.

Luminex 100 Analyzer Laser

The Luminex 100 IS system classifies per FDA 21 CFR 1040.10 and

1040.11 as a Class II laser product consisting of a Class I laser product (Luminex 100 analyzer) and a Class II laser product (barcode reader).

Figure 2-5 Laser Product Class Label

United States and international regulations require the following warnings to appear on the instrument during operation and maintenance.

This label appears on the back panel of the Luminex 100 analyzer.

Figure 2-6 Laser Radiation Caution Label

Under NO circumstances should you remove the Luminex 100 analyzer cover! When performing routine maintenance, turn power

to the Luminex 100 analyzer OFF and the disconnect the power cord.

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Luminex 100 IS User Manual Version 2.3 xMAP Technology

All laser apertures are located within the Luminex 100 analyzer and are contained within a protective housing. This label appears on the optics cover within the Luminex 100 analyzer.

Figure 2-7 Laser Class Label

Warning — Use of controls or adjustments or performance of

procedures other than those specified herein may result in hazardous radiation exposure.

Attention — L’utilisation des commandes ou réglages ou

l’exécution des procédures autres que celles spécifiées dans les présentes prescriptions peuvent entraîner d’une exposition à un rayonnement dangereux.

This label appears above the laser apertures located inside the optics enclosure inside the Luminex 100 analyzer.

Figure 2-8 Avoid Exposure Label

2 - 6 PN 89-00002-00-072 Rev. C

xMAP Technology Safety

Barcode Reader Laser This label is attached to the barcode reader.

Figure 2-9 Barcode Reader Laser Label

Do not stare into the beam or shine it into other people’s eyes.

Mechanical

Warning: During operation, this system contains exposed,

moving parts. Risk of personal injury is present. Observe all warnings and cautions.

Warning: During operation, this system contains exposed,

moving parts which could result in puncture hazard. Risk of personal injury is present. Keep hands and fingers away from the Luminex XYP instrument slot during operation.

Note: Access doors must be

closed while operating the Luminex 100 analyzer; the operator must be present during operation.

Biological

Warning: During operation, this system contains exposed,

moving parts which could result in pinch point hazard. Risk of personal injury is present. Keep hands and fingers away from the Luminex XYP instrument slot during operation.

Warning: Hum an and animal samples ma y cont ain biohazardo us

infectious agents. Where exposure (including aerosol) to potentially biohazardous

material exists, follow appropriate biosafety procedures and use personal protective equipment, such as gloves, gowns, laboratory coats, face shields, or mask and eye protection, and ventilation devices.

Observe all local, state, and federal biohazard handling regulations when disposing of biohazardous waste material.

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Heat

Warning: The heater plate of the Luminex XYP instrument may

be hot and could cause personal injury if touched. Do not touch the heater plate.

Blue Indicator Light

The blue light above the Luminex 100 analyzer sample arm indicates the on/off status of the Luminex 100 analyzer, and is harmless. The blue light-emitting diode (LED) does not emit light in the UV spectrum.

2 - 8 PN 89-00002-00-072 Rev. C

xMAP Technology Safety

Decontaminating the Luminex 100 Analyzer for Return Shipment

Note: It is the user’s

responsibility to decontaminate the analyzer before shipment.

Luminex Technical Support will give you a Return Material Authorization (RMA) number if they direct you to return the system. They will explain how to return the system according to Luminex procedures.

The accessible surfaces and the internal fluidics system must be sanitized and decontaminated before returning the analyzer. This is particularly important when biohazardous samples have been run. Make a copy of this page to fill out and return with the system.

Complete the following checklist, signed and dated, and return it with the Luminex 100 analyzer.

1. Remove all specimens, disposables, and reagents from the system.

2. Disconnect the sheath line going from the SD system to the analyzer.

3. Connect a sheath bottle filled with a solution of 10% to 20% household bleach and water to the analyzer.

4. Sanitize the system using the Sanitize command on the main screen of the system. Follow this by washing twice with distilled water.

5. Disconnect the system from AC power by turning off the power switch on the rear of the system, then unplugging the analyzer power cord from the wall source.

6. Disconnect the SD system and waste and sheath containers.

7. Rinse the waste container with 10% to 20% household bleach solution and drain.

8. Wash all exterior surfaces with a mild detergent, followed by a 10% to 20% bleach solution.

9. Open both front doors of the analyzer and clean all accessible surfaces with mild detergent followed by a 10% to 20% bleach solution.

10. Pack the system within a biohazard bag, place it in the corrugated box, then insert it in its original packaging or an approved shipping container. Attach this checklist to the top of the corrugated box prior to packaging in the crate.

Was there an internal leak in the system? Yes No

Print Name:

Signature:

Date: Instrument Serial No.

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2 - 10 PN 89-00002-00-072 Rev. C

The System

Theory of Operation Luminex 100 IS technology is based on flow cell fluorometry with

Luminex-developed innovations. The fluidics, optics, robotics, temperature control, software, and xMAP microspheres work together to enable simultaneous analysis of up to 100 analytes in a single test sample. Assay analysis requiring temperature control is provided through the Luminex XYP instrument heater block.

There are two fluidics paths in the Luminex 100 analyzer. The first path involves a syringe-driven mechanism that controls the sample uptake. This mechanism permits small sample uptake volumes from small reaction volumes. The syringe-driven system transports a specified volume of sample from a sample container to the cuvette. The sample is injected into the cuvette at a steady rate for analysis. Following analysis, the sample path is automatically purged with sheath buffer by the second fluidics path. This process removes residual sample within the tubing, valves, and probe. The second fluidics path is driven by positive air pressure and supplies sheath fluid to the cuvette and sample path.

Sheath fluid is the delivery medium of the sample to the optics component. The analysis sample is acquired using a sample probe from a 96-well microtiter plate via the Luminex XYP instrument and injected into the base of the cuvette. The sample then passes through with sheath fluid at a reduced rate resulting in a narrow sample core to ensure that each microsphere is illuminated individually. The sample injection rate is such that the xMAP microspheres are introduced to the optics path as a series of single events. The Luminex SD system lets you run samples continuously without refilling sheath bottles. It automatically draws sheath from a non pressurized bulk sheath container to constantly maintain a reservoir

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Luminex 100 IS User Manual Version 2.3 xMAP Technology

of pressurized sheath fluid. A single 20 liter sheath container provides enough fluid for 48 hours or more of normal operation.

The optics assembly consists of two lasers. One laser excites the dye mixture inside the xMAP microspheres and the second laser excites the fluorosphere bound to the surface of the xMAP microspheres. Avalanche photo diode detectors measure the excitation emission intensities of the color coding classification dye mixtures inside the xMAP microspheres and a photomultiplier tube detects the excitation emission intensity of the reporter molecule bound to the surface of the xMAP microspheres. High speed digital signal processors and advanced computer algorithms provide analysis of the xMAP microspheres as they are processed through the Luminex 100 analyzer. Results of the analyses are processed and provided in a report format.

Hardware The Luminex 100 IS system includes the following hardware:

• Luminex 100 analyzer

• Computer (PC), monitor, and accessories

• Luminex XYP instrument

• Luminex Sheath Delivery System (Luminex SD™)

• Power cables

• Alignment guide

• Two long sample probes

• Luminex XYP instrument sample probe

• Reservoir

•Shield

•Heater block

• Sheath fluid container

• Waste container

• Sheath fluid line

• Air line

• Sheath fluid intake line

• Communications: 1 serial communication cable

• Communications: 1 USB communication cable

• Communications: 1 CANBUS communication cable (short cable)

• Barcode reader

• Sample probe height alignment kit

• 3/32 Hexdrive, Balldriver wrench

3 - 2 PN 89-00002-00-072 Rev. C

xMAP Technology The System

xMAP Technology Reagents

Required Laboratory Reagents

Luminex 100 IS 2.3 Software

• Classification calibration microspheres (CAL1)

• Reporter calibration microspheres (CAL2)

• Classification control microspheres (CON1)

• Reporter control microspheres (CON2)

• Sheath fluid

• Household bleach

• 70% isopropanol or 70% ethanol

• Mild detergent

Luminex 100 IS 2.3 software provides complete control of the system and performs data analysis. Your Luminex 100 IS 2.3 system is preloaded with the Luminex software. However, we supply a software CD should you need to reinstall the software.

This software requires a dedicated system. Unauthorized additional software is prohibited and may result in improper operation of the system.

Luminex 100 IS Performance Specification

Speed • USB communications link for fast data transfer

• Automatic transfer of assay templates and new reagent information into the system via a large capacity read/write CD

• Installation: < 4 hours

• System calibration: < 10 minutes

• System controls: < 10 minutes

• Barcode reader entry of sample IDs

• Automatic post-analysis

• Analyze one 96-well plate/hour depending on manufacturer’s kit

• Up to 100 xMAP microsphere sets per sample

• Sheath flowrate: 90 µL/sec ± 5 µL

• Sample injection rate into detector area: 1 µL/sec ± 0.05 µL

• System warmup: 30 minutes. Systems that remain inactive for at least four hours will require a warm-up to restart the lasers. After

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acquiring sample, running system calibrators, running system controls, and warming up the instrument, the system resets the four-hour internal clock.

Accuracy and Precision • Sample uptake volume: ± 5%

• Classification of xMAP microspheres: > 80%

• Misclassification of xMAP microspheres: ≤ 2% - may vary by xMAP microsphere product lines. Refer to the specific product information sheet for further details.

• Temperature control: 0°C to + 2°C of target

• Internal sample carry over: < 0.9%

• Soluble background fluorescence emission at 575 nm automatically subtracted from fluorescence intensity values

Sensitivity • Detect 1000 fluorochromes phycoerythrin (PE) per xMAP

microsphere

• Reporter channel dynamic range: 3.5 decades of detection

Capacity The specifications below reflect minimum capacity values:

• 10 GB hard drive

• 100 MB read/write CD ROM

• Analyze multiple 96-well plates per batch

• Analyze multiple assay templates per plate

• Distinguish a minimum of 1 to a maximum of 100 unique xMAP microsphere sets in a single sample

• Detect and distinguish surface reporter fluorescence emissions at 575 nm on the surface of 1-100 unique xMAP microspheres sets in a single sample

• S ample core: 15-20 µm core at 1 µL/sec. sample inject rate

• Maintain samples at a constant temperature from 35°C to 55°C (95°F to 131°F)

• Automatic sampling from a 96-well plate

• Start sampling from any well position

• Sheath container and waste container hold enough volume to run up to two 96-well plates between refills

• M icrotiter plates with 96 wells must be compatible with the Luminex XYP instrument plate holder. The following microtiter plate types are compatible with the Luminex XYP instrument plate holder: flatbottom, conical, round, filter bottom, half plates [overall height no more than 0.75” (19 mm)], any color

• M icrotiter plates with 96 wells must be compatible with Luminex XYP instrument heater block temperature from 35°C to

3 - 4 PN 89-00002-00-072 Rev. C

xMAP Technology The System

55°C (95°F to 131°F) when performing heated assays and using the heater block

Luminex 100 Analyzer General

• Indoor use only

• Operating temperature: 15°C to 30°C (59°F to 86°F)

• Humidity: 20% to 80%, noncondensing

• Altitude: Operation up to 2400 m (7874 ft.) above mean sea level

• Physical dimensions: 43 cm (17 inches) W x 50.5 cm (20 inches) D x 24.5 cm (9.5 inches) H

• Weight: maximum of 25 kg (60 lbs.)

• UL installation category: UL Installation Category II, as defined in Annex J of UL 61010A-1

• Pollution degree: UL Pollution Degree 2, as defined in Section

3.7.3.2 of UL 61010A-1

• Shipping and storage: The allowable shipping and storage temperature and humidity ranges are 0°C to + 50°C and 20-80% noncondensing, respectively

• Input voltage range: 100 - 120 V~ ± 10%, 1.4 Amp, and 200-240 V~ ± 10%, 0.8 Amp, 47-63 Hz.

• AC inlet fuse: 3 Amp, 250 V~, fast acting

Optics • Reporter laser: 532 nm, nominal output 10-15 mW, maximum

500 mW, frequency-doubled diode; mode of operation, continuous wave (CW)

• Classification laser: 635 nm, 9.1 mW ± 6%, maximum output 25 mW, diode; mode of operation, continuous wave (CW)

• Reporter detector: Photomultiplier tube, detection bandwidth of 565-585 nm

• Classification detector: Avalanche photo diodes with temperature compensation

• Doublet discrimination detector: Avalanche photo diodes with temperature compensation

Fluidics • Sheath flow rate 90 µL ± 5 µL/second

• Cuvette: 200 micron square flow channel

• S ample injection rate: 1 µL/second

• Sample uptake volume: 20-200 µL

Electronics • Reporter channel detection: A/D resolution 14 bits

• Communications interface: USB

• L uminex XYP instrument, communications interface: RS 232

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Luminex XYP Instrument General

Luminex SD System General

• Ambient temperature: 15°C to 30°C (59°F to 86°F)

• Humidity: 20% to 80%, noncondensing

• Altitude: operation up to 2400 m (7874 ft) above mean sea level

• Physical dimensions: 44 cm (17.25 inches) W x 60 cm (23.5 inches) D x 8 cm (3 inches) H

• Weight: 15 kg (33 lbs.)

• UL installation category: UL Installation Category II, as defined in Annex J of UL 61010A-1

• Pollution degree: UL Pollution Degree 2, as defined in Section

3.7.3.2 of UL 61010A-1

• Heater operating range: 35°C to 55°C (95°F to 131°F) with tolerance 0°C to +2°C

• Input voltage range: 100-240 V~ ± 10%,1.8 Amps, 47-63 Hz

• AC inlet fuse: 3 A, 250 V~, fast acting

• Ambient temperature: 15°C to 30°C (59° to 86°F)

• Humidity: 20% to 80%, noncondensing

• Altitude: designed to operate at up to 2400m (7874 feet) above mean sea level

• Physical dimensions: 20 cm (8 inches) W x 30 cm (11.75 inc he s ) D x 24.75 cm (9.75 inches) H

• Weight: 9 kg (20 lbs)

• UL installation category: UL Installation Category II, as defined in Annex J of UL 61010A-1

• Pollution degree: UL Pollution Degree 2, as defined in Section

3.7.3.2 of UL 61010A-1

• Input voltage range: 100-240 V~ ± 10%, 0.4 Amps, 47-63 Hz

• AC inlet fuse: 2 Amp, 250 V~, time lag

PC Specifications For systems using a PC, A Dell OptiPlex GX280 or Dell Optiplex

GX520 (or newer PC) is shipped with the Luminex 200 system. For systems using a laptop, a Dell D610 Notebook is shipped with the system. Microsoft The power requirements are 115-230 V~, 6 Amps, 50-60 Hz

For updated information regarding the PC, notebook, or operating system, go to http://www.luminexcorp.com, then click on the Support link to open the FAQ list.

3 - 6 PN 89-00002-00-072 Rev. C

® Windows® XP is pre-installed on the computers.

xMAP Technology The System

Recommended Additional Equipment

Uninterruptible

Power Supply

(UPS)

Surge Protector If you do not use a UPS, use a surge protector. Choose a protector

Printer Printer, HP LaserJet 2300 or available equivalent

Barcode Labels Use the Code 128 barcode label type when scanning barcode labels

Vortex VWR product number 58816-121: Speed range 0-3200 rpm or

Luminex highly recommends using an uninterruptible power supply (UPS) to protect your system from power outages. Choose one that can provide 1050 Watts for at least 45 minutes. The UPS should be UL listed, CSA certified, and CE marked when used internationally.

that meets your needs. Factors to consider include electrical environment, endurance, suppressed voltage rating, and method of protection. It should have six outlets, rated at least 1500 Watts, and be UL listed, CSA certified, CE marked for nondomestic use when used internationally .

into the system as patient identities.

equivalent

Bath Sonicator Cole-Parmer® product number 08849-00: Operating frequency 55

kHz or equivalent

System Overview The system consists of three subsystems: electronic, fluidic, and

optical. The following section describes the user-accessible components of each subsystem.

Electronics

Power Input

Module

Communications

Ports(SB9-PIN)

Luminex 100

Analyzer

Ventilation Filter

The power input modules contain the on/off switch and fuses.

The communications port connects the Luminex 100 analyzer or the Luminex XYP instrument to the computer, and the Luminex SD system to the Luminex 100 analyzer.

Located on the bottom of the Luminex 100 analyzer, the filter must be checked and cleaned as necessary. For proper ventilation, do not obstruct the area below and allow at least two inches (5 cm) of clearance around the Luminex 100 analyzer.

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Luminex XYP

Instrument

Ventilation Filter

The XYP instrument ventilation filter cleans the air that cools the internal parts of the Luminex XYP instrument. See Figure 3-1.

1. Air intake filter access door 5. Communication Ports (DB9)

2. Power Switch 6. XYP Communication Port (DB9)

3. Power Input Module 7. Analyzer ventilation filter

4. XYP Ventilation Filter

5 7

(on bottom of analyzer)

Fluidics

Figure 3-1 Back of the Luminex 100 Analyzer and Luminex XYP

Instrument

Sample Arm The sample arm transports the sample from the sample tube to the

cuvette. The carriage drops to the microtiter well for sample retrieval.

Luminex XYP

Instrument Sample

A stainless steel sample probe acquires the sample. A shorter probe is provided for shipping and troubleshooting.

Probe

Warning: During operation, this system contains exposed moving

parts that can result in a puncture hazard. Risk of personal injury is present. Keep hands and fingers away from the sample probe. The shield should be in place.

Cheminert® Fitting This fitting attaches the Luminex 100 analyzer sample arm tubing to

the sample arm. Disconnect this fitting when you remove the sample probe. See Figure 3-2.

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xMAP Technology The System

The alignment guide directs the sample probe into the Luminex XYP instrument.

1. Cheminert Fitting

2. Sample Arm

3. Luminex XYP Instrument Sample Probe

4. Shield

5. Alignment Guide

Figure 3-2 Cheminert Fitting

Access Doors The Luminex 100 analyzer has three access doors. Two of the access

doors are on the front, and the third is on the back. The front left access door supplies access to the sheath filter. The front center access door supplies access to the syringe. The rear access door supplies access to the air intake filter. See Figure 3-3 and Figure 3-4.

1. Left door, access to service panel 2. Center door, access to syringe

Figure 3-3 Luminex 100 Analyzer Access Doors

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Air Intake Filter A replaceable air intake filter cleans the air used to pressurize sheath

fluid. This filter is enclosed behind an access door located on the back of the Luminex 100 analyzer. See Figure 3-4.

Figure 3-4 Air Intake Filter

Syringe The syringe delivers a sample from the 96-well microtiter plate to the

cuvette. See Figure 3-5.

1 2

1. Syringe Seal 2. Syringe

Figure 3-5 Syringe and Syringe Seal

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xMAP Technology The System

Sheath Filter The sheath filter removes particles greater than ten microns in

diameter from the sheath fluid. See Figure 3-6.

Figure 3-6 Sheath Filter

Air, Waste, and

Sheath Fluid

Connectors

The air, waste, and sheath connectors, located on the left side of the analyzer, connect to the SD system and waste fluid containers using clear tubing. The air connector is green, the sheath fluid connector is blue, and the waste fluid connector is orange. See Figure 3-7.

1. Sheath fluid connector (blue)

2. Air connector (green)

3. Waste connector (orange)

Figure 3-7 Air, Waste, and Sheath Fluid Connectors

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Luminex Sheath

Delivery System

Note: If you are not using the SD

system, sheath fluid levels must be monitored manually. Check the sheath fluid level before st ar ting a run or procedure.

Waste Fluid

Container

For proper operation, place the Luminex SD system at the same level as the base of the Luminex XYP instrument. Do not put it on top of the Luminex 100 analyzer.

Warning: If biological samples have been tested with the

system, use your standard laboratory safety practices.

The waste fluid container receives waste from the system. The waste container should not be placed on top of the instrument.

Caution: Waste levels must be manually monitored. Do not

allow the waste container to overflow!

Optical The optical system consists of the optical assembly and the excitation

lasers. The optical assemblies do not require manual adjustment by the user.

xMAP Technology Reagents

The xMAP technology reagent system consists of classification calibration microspheres, reporter calibration microspheres, classification control microspheres, and reporter control microspheres.

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Basic Concepts

Background Information

xMAP technology is a versatile system that measures soluble analytes. The Luminex 100 IS system performs simultaneous, discrete measurements of multiple microsphere-based reactions from a single specimen aliquot. For more conceptual information, refer to Practical Flow Cytometry, 3rd edition, by Howard M. Shapiro, M.D. (New York: Wiley-Liss Inc., 1995).

Fluidics There are two fluidic paths in the Luminex 100 analyzer. The first

path involves a syringe-driven mechanism that controls the sample uptake. This mechanism permits small sample uptake volumes from small reaction volumes. The syringe-driven system transports a specified volume of sample from a sample container to the cuvette. The sample is injected into the cuvette at a steady rate for analysis. Following analysis, the sample path is automatically purged with sheath fluid by the second fluidics path. This process effectively removes residual sample within the tubing, valves, and probe. Approximately 160 µL of sheath fluid is dispelled into each well following sample acquisition. The second fluidics path is driven under positive air pressure and supplies sheath fluid to the cuvette and sample path.

Excitation The excitation system in the Luminex 100 analyzer involves two

solid-state lasers. A reporter laser excites fluorescent molecules bound to biological reactants at the xMAP microsphere surface, and a classification laser excites fluorochromes embedded in the xMAP microsphere. The lasers illuminate the xMAP microspheres as they flow single-file through the cuvette. RP1 refers to the excitation wavelength. CL1 and CL2 refer to the dyes embedded in the microsphere. DD refers to the channel that discriminates against doublets based on size.

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Photodiodes and a photomultiplier tube receive fluorescent signals from xMAP microspheres. The Luminex 100 analyzer digitizes the waveforms and delivers the signals to a digital signal processor (DSP). Proprietary algorithms function with the DSP to greatly increase sensitivity.

xMAP Microspheres The xMAP microspheres are highly uniform, polystyrene particles

that have been crosslinked during polymerization for physical and thermal stability. Varying amounts of fluorochromes embedded within each xMAP microsphere give each xMAP microsphere set an unique fluorescent signal. To ensure the stability of this signal, it is essential to protect the microspheres from light. Follow the product information sheet instructions for storage procedures for xMAP microspheres and assay kits.

Calibrator xMAP microspheres are used to normalize the settings for the reporter channel, both classification channels and the doublet discriminator channel for the Luminex 100 analyzer.

The control xMAP microspheres are used to verify the calibration and optical integrity of the system.

Software Overview This section provides a brief overview for using the Luminex 100 IS

2.3 software.

With the Luminex 100 IS 2.3 software, you work with assay kits provided by a kit manufacturer. A template may be included with each kit that is imported into the software. The template includes a sequence of commands required for the assay.

Once you select the template, you enter sample data into a “batch.” Sample input can be done quickly with either the keyboard or a barcode scanner. A batch can include as many samples as you have for the assay and can include multiple microtiter plates. You can even group multiple batches together into a multi-batch for efficient processing. You can process a batch immediately or choose to archive the batch for testing later.

As testing proceeds through the plate, you will see the display update to show the wells that have been processed. Progress is shown in both graphic and tabular form.

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System calibrators and controls are provided as part of the system. Calibrate at least once monthly, when the delta calibration (dCAL) temperature changes by ±3 degrees, or if the system is powered off or moved. You must run system controls following calibration and as often as you like to ensure that your system continues to operate optimally.

The software provides you with a variety of reports.

• Analyte Reports provide batch results grouped by the test in a batch.

• Patient Summary reports provide batch results grouped by each patient or unknown in a batch.

• Clinical Patient reports provide a breakdown of samples according to the test analysis with that sample.

• Maintenance Reports provide a history of all maintenance operations performed during a specified period of time.

• Calibration Trend reports provide results of calibration commands performed over a specified time period.

• System Control Trend Reports provide results of system controls performed over a specified time period.

• Batch Summary Reports provide batch information in a sample versus test grid format.

• Quality Control Reports track the trends of assay controls over a period of time

These reports let you look at specific details regarding the samples you process through the Luminex 100 IS 2.3 software and system operation.

System tools help you monitor the system including a display showing real-time system property values and a message log. Errors are shown in the message log and also on your reports. You can even add comments to specific results in some of these reports.

To keep track of your reagents, the Lum inex 100 IS 2.3 software stores lot numbers, expected values, and expiration information.

A comprehensive set of self-diagnostic tests ensure that your system is working correctly. These tests are performed at startup and can be performed at any time. If any problem is detected, an error message appears in the message log to inform you.

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Using Luminex 100 IS 2.3 Software

This manual describes how to use the Luminex IS 2.3 software. It includes a glossary and information regarding the CSV file setup. The software user should have a basic familiarity with computers and Microsoft® Windows®. Commands are often available through more than one method, such as from the main menu bar, the toolbar, or on different windows. However, each procedure in this manual will describe only one method of accessing commands.

This chapter has the following sections:

• Main Window - This section describes the main window of the Luminex IS 2.3 software, including tabs and commands. Use this section to become familiar with the main functions of the software.

• S econdary Windows - This section describes the secondary windows in the Luminex IS 2.3 software, including the Analysis Window and Batch Setup Window.

• Commands - This section describes the functions of the commands in the Luminex IS 2.3 software.

• P rocedures - This section describes how to perform tasks using the Luminex IS 2.3 software.

When using microtiter plates, use plates with wells that will hold at least 185 µL (the extra 25 µL from the sample, plus an extra 160 µL that is dispensed back into the well following acquisition).

Main Window The Luminex IS 2.3 software starts automatically when you log into

Windows.

When the first software User window opens, the Main window is open, with the Home tab displayed, as shown in Figure 5-1

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1. Title bar 4. Tabs

2. Menu bar 5. Status bar

3. Tool bar

Figure 5-1. Luminex IS 2.3 Main Window

There are five major parts in the Main window: Title bar, Menu bar, Tool bar, Tabs, and Status bar. A brief description of each of these components is shown below.

The Title Bar displays the name of the software.

The Menu Bar contains three menus, File, Tools, and Help menu. A more thorough description of the Menu is shown on page 5-3.

The Toolbar - has shortcut buttons for frequently-used commands. A more thorough description of the Toolbar is shown on page 5-4.

There are five Tabs, organized by function. The tab that displays when the software starts up is the Home tab. A more thorough description of each of these tabs begins on page 5-4.

The Status bar displays the system status at the bottom of the main window. A more thorough description of the Status bar is shown on page 5-17.

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Menu Bar The menu bar contains the following menus: File Menu, Tools

Menu, and Help Menu.

File menu - contains the following commands:

• Import Template

•New Batch

• Open Batch

• Delete Batch

• Edit Patient List

• Open Incomplete Batch

• Batch Comment

• New Multi-Batch

• Open Multi-Batch

• Data Analysis

• Export Batch Data

• Print Report

•Exit

Tools menu - contains the following commands:

• Connect

• Disconnect

• Database Backup

• Database Restore

• Erase Database

• Update CAL Targets

• Update CON Targets

• New Assay Lot

• Options

•Cleanup

For information on each of these commands, see the Commands section, beginning on page 5-31.

Help menu - this contains menu selections that open the online help and describe the software and hardware.

• Contents opens the online help

• About the Device opens a dialog box that shows the version

and serial numbers of the Luminex instruments you are using.

• About the Software opens a dialog box that shows the

version of the Luminex Software you are using.

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Toolbar The shortcut buttons on the toolbar can be found in other locations in

the software. The Help button opens the online help. The Single Step option on the toolbar allows you to pause the system in between each command or sample acquisition within a batch.

1. Import Template 8. Print Report

2. New Batch 9. Connect to the Instrument

3. Open Batch 10. Disconnect from the Instrument

4. Create New Multi-Batch 11. Eject/Retra c t

5. Open Multi-Batch 12. Open Help File

6. Start Analysis 13. Single Step

7. Export Batch Data

Figure 5-2. Luminex IS 2.3 Toolbar

Tabs This section describes the tabs in the Main window.

Home tab This is the default tab. It is organized by the order of system use. It

contains a Favorites list and five button groups representing data acquisition and maintenance categories.

1. Favorites List 2. Button Groups

Figure 5-3. Home Tab

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Favorites. You can use the Favorites list to create a customized list of often-used commands and templates. Items appear in alphabetical order for easy locating. For more information on setting up the Favorites list, see page 5-42.

Button Groups. Button groups are grouped according to function.

•The Daily Startup group contains the Warmup, Prime,

Alcohol Flush, and Wash Commands. For more information on daily startup, see page 5-43. For more information on the commands, see the Commands section on page 5- 31.

•The Instrument Calibration group contains the CAL1,

CAL2, CON1, and CON2 buttons. For more information on calibrating the system, see page 5-46. For more information about these commands, see the Commands section on page 5-

31.

•The Batch Setup group contains the New Batch, New

Multibatch, and New Lot buttons. For more information on creating and running batches and multibatches, see page 5-51. For more information about these commands, see the Commands section on page 5-31.

•The Reports and Analysis group contains the Analysis and

Print buttons. For more information on running reports and analyses, see page 5-87. For more information about the Analyses and Print commands, see the Commands section on page 5-31.

•The Daily Shutdown group contains command buttons that

you can use when shutting down the system. For more information on performing a daily shutdown, see page 5-97. For more information on the Sanitize and Soak commands, see the Commands section starting on page 5-31.

Run Batch Tab The Run Batch tab contains a microtiter plate and reservoir image,

XYP and DD temperature readbacks, a sheath pressure gauge, print button, command list displaying batch commands and their status, and plate and XYP command buttons. See Figure 5-4.

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1. Microtiter plate/reservoir image 2. Temperature and Pressure Gauge s

3. Command List

Figure 5-4. Run Batch Tab

The command buttons on the Run Batch tab are:

• Print Worklist • Eject/Retract

•Start Plate •Resume

• Cancel Command • Pause

• Cancel all

For more information on these commands, see the Commands section, starting on page 5-31. For more information on setting up batches, see See “Batch Setup Procedures” on page 51..

The microtiter plate and reservoir image represents where you place samples or other fluids used in running or maintaining th e system. Samples are analyzed vertically, from top to bottom within the column, and then from left to right for subsequent columns.

The Temperature and Pressure Gauges show information about the analyzer and the XYP instrument. The XYP heater temperature measures the internal Luminex XYP instrument plate temperature.

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The DD temperature measures the doublet discriminator temperature. DD temperature shifting indicates a need for system recalibration. If you have not calibrated, the arrows showing the temperature range for the DD temperature both appear at the bottom of the thermometer, and the thermometer appears in red. An out-of-range temperature logs an error, but does not halt the acquisition.

For information on setting the XYP instrument heater temperature, see page 5-45.

The Command List displays commands associated with batches, new advanced batches or multi-batches loaded for processing on the system. The Command List shows the status of each command as the system processes it, and whether the command completes successfully or fails. Figure 5-5 shows that the first two commands were successful, the third command is running, and the rest of the commands are pending. Figure 5-6 shows that the third command failed.

Figure 5-5. Command List Section of the Run Batch Tab

Figure 5-6. Command Failure Notation in the Command List

Errors are recorded in the Message Log on the Diagnostics tab. See page 5-9 for more information about the Diagnostics tab. Doubleclick on failed commands in the Message Log for additional information. You can right-click a highlighted row to copy data to the clipboard or clear the batch from the screen.

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Maintenance Tab Use the Maintenance tab to perform maintenance, XYP, daily

startup, calibration, and verification commands to maintain your system and keep it in optimal working order.

Figure 5-7. Maintenance Tab

See the Commands section starting on page 5-31 for detailed information about these commands:

•Warmup

•Prime

•Backflush

• Alcohol Flush

• Sanitize

•Wash

•Drain

•Soak

• Self Diagnostics

• Calibration Commands: CAL 1, CAL2, CON 1, CON2, New CAL target and New CON target

• Sample Probe Down

• Eject/Retract

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Instructions for maintenance procedures begin on page 5-93. Table 511 on page 5-93 shows a recommended use schedule for maintenance operations.

Diagnostics tab Use the Diagnostics tab to monitor the progress of commands you

initiate in the system. Features on this tab monitor the state of system components. This tab displays the System Monitor, Detailed Sample Progress chart, and Message Log. The Text on the Diagnostics tab turns red if the system encounters an error. The Message Log on the Diagnostics tab indicates where the error occurred.

Figure 5-8. Diagnostics Tab

Use these tools to find information about the system and what occurs during sample acquisition and other functions. For example, you may look on the Message Log to see the last completed command or the one currently in progress.

The System Monitor provides information about the physical state of the Luminex analyzer including lasers and system calibration status. The values displayed are reported directly from the Luminex analyzer and the Luminex XYP instrument. It shows whether the calibration or control results completed successfully by displaying green text for successful events and red text for failed events.

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The system diagnoses real-time system problems related to fluidics and optics. The system also detects and reports if the Luminex XYP heater block temperature is out of range or experiencing unacceptable temperature conditions, including these items:

• L uminex XYP heater block temperature time-out

• heat circuit failure

• t emperature out-of-range sensing

• t emperature change since calibration, or a change in channel temperature

Table 5-1 defines the values listed in the System Monitor. These values are useful for diagnostic purposes when communicating with Luminex Technical Support.

Table 5-1. System Monitor Values

Property Value Units of Measure

Air Pressure PSI, air pressure to the sheath

container from the air pump

Sheath Pressure PSI, sheath pressure from the

sheath container through the system

Delta Cal Temperature °C, temperature deviation from

the last calibration

Board Temperature °C, temperature of the analog

board

DD Temperature °C, DD temperature at the U

block inside the optics platform

CL1 Temperature °C, CL1 temperature at the U

block inside the optics platform

CL2 Temperature °C, CL2 temperature at the U

block inside the optics platform

XYP Board Temperature °C, temperature of the XYP

board inside the Luminex XYP instrument

XYP Heater Temperature °C, temperature of the XYP

heaters inside the Luminex XYP instrument

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Table 5-1. System Monitor Values (Continued)

Property Value Units of Measure

XYP Heater Temperature In Range

Indicates if the XYP heater is in the set range

The Detailed Sample Progress section displays the percentage of completion for each bead ID or test. The graph shows real-time progress, so that as each sample is analyzed, the graph adjusts to show progress. Use the zoom button to view up to 20 tests at a time. Use the toggle button to view the bead ID or test name. Use the Expand Margin (up/down and left/right) arrows to expand the chart margins and view longer test names. See Figure 5-9.

1. Zoom button

2. Toggle button

3. Expand Margin arrows

Figure 5-9. Detailed Sample Progress

The Message Log is the lower pane on the Diagnostic tab. It displays a list of completed commands, errors, and warnings. It also displays command progress, time and date, and results. See Figure 5-10.

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Figure 5-10. Message Log

The log displays actions in color-coded text and shading. Items in the Message Log appear in the following color codes:

• Green text represents a successful system calibration, verification command, acquisition, or maintenance functions.

• Red text represents failed commands, errors, or warnings.

• Black text represents normal processes and actions.

• Yellow shading indicates that a detailed description about the processes or actions is available. This color may vary depending on your tool tip color system settings.

Acquisition Detail

Tab

To view details of a message, double-click the shaded row. A dialog box opens providing details. To clear the Message log, right-click in the Message Log area, and click Clear on the menu.

The Acquisition Detail tab offers advanced batch sample monitoring and “on the fly” data acquisition without templates. The primary function is real-time monitoring of batch sampling during acquisition through a display of sample bead statistics, histogram, and dot plot data.

During batch acquisition, bead statistics can be useful if batch errors occur. For example, if samples are constantly failing due to insufficient bead count, you can monitor whether the failure is due to low bead concentration or if other assay problems are present.

The Acquisition Detail tab provides access to the Developer Workbench (DWB) software features. Details about these features are provided in the Luminex Developer Workbench Guide Version

2.3 documentation. You must have the DWB software installed to

enable these features.

Caution: Do not alter kit manufacturer’s predefined templates or

create alternative templates for off the shelf kits unless instructed by the kit manufacturer.

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Acquisition Detail tab features:

• Batch Name and Description

• Batch Data Area

• Acquisition Detail Toolbar

• Histogram

• Dot Plot

Figure 5-11 identifies the major features of the Acquisition Detail tab.

1. Luminex IS 2.3 T o olbar 5. Status Bar

2. Batch Name and Description 6. Histogram

3. Batch Data Area 7. Dot Plot

4. Acquisition Detail Toolbar

Figure 5-11. Acquisition Detail Tab

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The Batch Name and Description section displays the name of the batch as well as its description.

The Batch Data area displays sample results. The left column shows plate location and Sample ID description. The remaining columns display selected bead sets for the assay. Each row represents the data for each bead set from one well.

The Acquisition Detail Toolbar shown in Figure 5-12 provides functions to acquire raw data without using templates. The toolbar has additional buttons if you have Developer’s Workbench installed. For more information about the commands on the Acquisition Detail Toolbar, see the Command section, beginning on page 5-31.

1. Replay Batch 6. Cancel All

2. New Advanced Batch 7. Eject/Retract

3. View Batch Data 8. Pause

4. Start Plate 9. Resume

5. Cancel Command

Figure 5-12. Acquisition Detail Toolbar

The Histogram, in the lower left section of the Acquisition Detail tab, defaults to display the Doublet Discriminator (DD) on the X axis and Events on the Y axis. Doublets appear when two microspheres stick together, creating undesired results. When you enable the gate, two vertical red, dashed lines appear to represent gate positions determined by the template or settings established within the New Advanced Batch. After setting the gate, everything outside the gate is ignored. You cannot change the gate position during batch acquisition. You can change the gate positions before data acquisition, after the system finishes acquiring data, or after pausing the system. The change is visual. The data is still collected according to the gate positions set in the assay template or New Advanced Batch setup. The gate in effect when the system collects data determines which values to use to obtain the result. Applying a gate or changing a gate for existing data does not change your calculated values. The gate positions, also located in the lower corner of the histogram, are used while collecting data are the numerical values selected in the template or the New Advanced Batch.

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1. Gate Boundaries 2. Aggregate Beads 3. Numerical Gate Position

Figure 5-13. Set DD Gate Example

The Histogram contains the Show Bead menu and four buttons:

• Autoscale

•Zoom

• Log/Linear

• Maximize

For more information on these commands, see the Commands section beginning on page 5-31.

The Dot Plot (or bead map) appears in the lower-right section of the Acquisition Detail tab. See Figure 5-14. The dot plot shows a graphical display of real-time data collection.

Luminex recommends using the default settings to collect data. The default axes are Classification 1 on the X axis and Classification 2 on the Y axis. To see the dot plot, you must use the default axis. To display the bead set information, hover the mouse pointer over the desired region. You can change the X axis and Y axis of the dot plot for troubleshooting purposes, although you should use the default settings in all other scenarios.

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Figure 5-14. Dot Plot Display Example

Four buttons appear at the top of the frame to control the display:

• Density/Decaying

• Log/Linear

•Zoom

• Maximize

You can toggle between two types of dot plots using the Density/ Decaying button. The Decaying Dot Plot plots only the 100 mostrecent acquired events. The Density Dot Plot displays a constant accumulation of events. Increasing density is indicated by contrasting colors. See Table 5-2 for the density dot plot color legend.

Table 5-2. Dot Plot Color Legend

Layer Color

0 1 2 3 4 5 6 7 8

The density dot plot allows visual elimination of data values determined to be insignificant to the display. Luminex recommends you collect your data in density dot plot mode to observe all

none dark blue pink dark green cyan light blue light green orange dark red

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collected events. Post acquisition does not display decaying dot plot; it’s only a real-time function.

Status Bar The Status Bar displays information about the Command State,

Device Status, Device Activity, Laser Status, Total Events per Second, and Region Events per Second. Color coding indicates the urgency of each item’s status. Device Activity uses no color coding.

Figure 5-15. System Status Bar

Table 5-3 describes the types of status bar messages in relation to the message color coding.

Table 5-3. Status Bar Color Coding

Category Color Indicates

Command State

Indicates communication status of the Luminex analyzer or operations being processed

Green Idle or processing Yellow Connecting, pausing, or paused Red Disconnected or locked out

Device Status Indicates the current process of or warning about the

Luminex analyzer

Green Running or standby Yellow Busy, pressurizing, sheath is empty, or warming

Red Not ready or disconnected

Device Activity

Indicates the activity that the Luminex analyzer is performing. Note: The device activity has no color code.

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Table 5-3. Status Bar Color Coding (Continued)

Category Color Indicates

• Idling: waiting for a command

• Aspirating: drawing in sample

• Collecting Data: collecting data

• Sanitizing: sanitizing the instrument

• Washing: washing the sample line

• Priming: priming the instrument

• Calibrating: calibrating the instrument

• Canceling: canceling a command

• Backflushing: backflushing the instrument

• Draining: draining the instrument

• Warming Up: warming up the instrument

• Verifying: verifying calibration

• Soaking: soaking the probe

• Adjusting sample probe

• Self Diagnosing: performing self-diagnostic routine

Laser Status Laser temperature and readiness status

Green Lasers warmed up Yellow Warmup timer countdown in seconds from

1800

Red Lasers off

Total Events/

Number of total bead events detected per second

Second Region

Events/Second

Number of bead events detected per second that are classified in a region

The Status Bar displays status information as the software processes commands. Table 5-4 shows the typical messages that appear in the status bar. Additionally, information displays as text on the Diagnostics tab.

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Table 5-4. Status Bar

Communication Message Details

Status Bar

Message Indicates

Color

Green Idle Waiting to process the next

command.

Standby The Luminex analyzer is ready

and waiting to perform a command.

Yellow Connecting The software is attempting to

connect to the instrument.

Processing The instrument is communicating

with the software as it processes commands.

Pausing The instrument has stopped

processing the list of commands, but finishing the active command.

Paused The software stopped processing

the list of commands. A “resume” function becomes available to change the state back to “processing.”

Busy The instrument is processing a

maintenance command.

Red

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Disconnected

Locked Out Another application currently has

The software has not yet attempted to connect or fails to connect to the instrument.

control of the instrument. The software locks out as long as the other application runs. To remove the locked out status, close the other application or wait until the application completes.

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Secondary Windows

In addition to the main window, there are other windows that are displayed when you select certain commands. These are the Batch Setup Window and Analysis Window. Additional windows display in the Developer’s Workbench software, if you have it installed on your system. For more information about the Developer’s Workbench, see the Luminex Developer Workbench Guide for Version 2.3.

Batch Setup Window In the Batch Setup window, you define information used to create

batches. All of the commands are on the same page; there are no tabs. The window opens when you click New Batch and select a template.

The following features are part of the Batch Setup Window:

• Batch Info group box

• Template Info group box

• Insert command

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• Command list

• Microtiter Plate image

• Command button group box

• Save group box

• Standard Info group box

• Control Info group box

• New Lot button

The Batch Info group box has text boxes in which you can input the name, description, and creator name of a new batch.

The Template Info group box shows the name, description, version number, and developing company for the template that you are using in a batch.

The Insert command lets you insert a patient into a batch or skip a well.

The Command list displays the commands that were imported from the template, plus commands added using the Insert command or manually entered. The lock in the far left column indicates that the command was imported from the template and cannot be changed in the Batch Setup window.

The Microtiter Plate image reflects the command information for each well, as defined in the template.

The Command button group contains the Finish, Cancel, Load Pa List, and Help command buttons. For more information on these commands, see the Commands section, beginning on page 5-31.

The Save group box contains the Save and Load and Save only option buttons. For more information on these commands, see the Commands section, beginning on page 5-31.

The Standard Info group box is visible only if the template you are using requires the use of standards. If standards are used in the template, the Standard Info group box reflects the Standard information as defined in the template product.

The Control Info group box is visible only if the template you are using requires the use of controls. If controls are used in the template, the Control Info group box reflects the Control information as defined in the template products.

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The New Lot button is visible only if the template you are using requires the use of standards or controls. The button opens the Update Lot Information dialog box, in which you can manag e lot information.

Analysis Window When the system analyzes batches, it displays the data in a three-tab

format within the Analysis window. The following three tabs present the batch information in greater detail:

• E rrors tab—lists errors that occur during batch acquisition, such as controls that failed.

• Standards tab—lists all tests in the batch, a regression chart for each test, and the standards or controls associated with the batch.

• S amples tab—lists background samples and all sa mples or unknowns acquired in the batch with either a qualitative or quantitative result.

Nine function buttons are available on the lower portion of the Analysis window. These buttons perform tasks or functions that are relevant to the Standards tab although they appear on all three tabs of the Analysis window. The buttons are:

• Next Test (F2)

• Previous Test (F3)

• Invalidate Standard (F4)

• Validate Standard (F5)

• Invalidate Control (F6)

• Validate Control (F7)

• Change Standard - only if Developer’s Workbench is installed.

• Change Lot (Alt + F8)

• Recalc (Standards tab only) - only available on the Standards tab, and only if the Auto checkbox is not selected.

For a description of these commands, see “Commands” on page 5-

31. For information on procedures using these commands, see

“Validating or Invalidating Standards and Controls” on page 5-84 and “Change Lot” on page 5-85.

In addition to the function buttons, you can choose to sort data sequentially by order acquired, or alphabetically by Sample ID. These options are available in the Sort group box next to the function buttons. These sorting functions are helpful in viewing batches containing replicate samples. Replicate samples are defined as samples with identical sample identifications. Replicate standards,

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controls, and unknown samples are not always acquired in sequential wells and thus make sample viewing difficult. When viewing samples alphabetically all replicate samples are displayed together with the replicate sample’s average (AVG), then individual samples. If you view replicate batches sequentially, each sample displays in the order it was acquired, followed by a replicate average summary section.

Errors Tab The Errors tab displays a list of errors that occurred during

acquisition. It organizes the errors in two categories: System and assay errors (top section of tab), and Sample errors (bottom section of tab) Table 5-5 lists the errors in each category

Figure 5-16. Analysis Window - Errors Tab Open

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Table 5-5. Error Categories

System and Assay Errors Sample Errors

• Instrument Not Calibrated

• Failed Verification (system lists each failed control)

• Failed Control (verifier failed)

• Temperature Divergence from Calibration Temperature

• Failed Curve Fit

• Analysis Error

• APD T emp Range Exceeded

• XYP Temp Unstable

• Low Voltage

• High Sheath Pressure

• Low Sheath Pressure

• Command Timeout

• Low Laser Power

• Cannot Calculate Inverse Function

• Failed Std in Batch

• Insufficient Bead Count

• Sample High/Low

• Analyzer Error

• High Sheath Pressure

• Low Sheath Pressure

• Sample Timeout

• Sample Empty

• Cannot Calculate Inverse Function

• Different Qualitative Results

Standards Tab The Standards tab lists all the batch tests with system comments

specific to each test. The detailed test information that is displayed on the three tabs of the Analysis window is determined by the test selected under the Standards tab.

The Standards tab displays the quantitative batch’s standard curve. It displays qualitative batches as a graph plotting the assay standard. Above the graphical information, the tab displays the formula that it used to calculate batch data. Below the graphical information, the tab displays the Standards and Controls values for the selected test.

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Figure 5-17. Analysis Window - Standards Tab Open

When a median fluorescence intensity (MFI) value for an unknown sample or control lies outside the standard curve MFI range, the concentration is not calculated. The unknown sample or control result is reported as less than (<) or greater than (>) next to the corresponding standard expected concentration. The out-of-range samples and controls will also have a “Sample High/Low” statement in the comment column. If a sample lies within the standard curve MFI range, but the sample’s MFI value does not intersect the curve, the result will be reported as “Error”. A “cannot calculate inverse function” statement displays in the comments column. This error condition usually occurs when a standard curve “flattens out” at the high or low end. Examples of out-of-range sample labeling for noncompetitive assays are shown in Table 5-6. Figure 5-18 shows what a standard curve for a non-competitive assay should look like.

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Table 5-6. Non-Competitive Out-Of-Range Labeling

Condition Concentration Label MFI of Standard Referenced

Left of Curve < min concentration standard Lowest

Right of Curve > max concentration standard Highest

Figure 5-18. Non-Competitive Assay

Examples of out-of-range sample labeling for competitive assays are shown in Table 5-7. Figure 5-19 shows what a standard curve for a competitive assay should look like.

Table 5-7. Competitive Out-Of-Range Labeling

Condition Concentration Label MFI of Standard Referenced

Left of Curve < min concentration standard Highest

Right of Curve > max concentration standard Lowest

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Figure 5-19. Competitive Assay

The standards tab displays an Expected Concentration column for standard and control samples. The Standards Expected Concentration column allows users to edit the standard concentrations. You can edit controls; however, you must select the control name or Expected Concentration box for that control, then click Change Lot.

When editing standard and control lot values the system may prompt you for a new number. If this is the first batch you analyze using this lot number, the system allows editing without requiring a new lot name. However, if you have analyzed a previous batch using this lot, the system will require that you rename the lot if edited or modified.

Background samples are commonly used in the assay process. Although recommended, the discretion on whether to use or not is left to the assay or kit developer. If background samples are included they are defined in the assay template.

Background samples are samples with no active test substance. The system uses background samples to remove assay background noise from the sample results. Typical background sample s contain coupled beads,

assay buffer, detection reagents, and reporter

fluorophore. Background samples do not contain target analytes.

If the background sample is designated accordingly, the system subtracts the background sample’s reported fluorescence from all other samples in a batch to report net MFI. If a batch contains more

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than one background sample, the fluorescence values of the two background samples are averaged together and subtracted from the other samples to obtain a net fluorescence intensity.

Samples Tab The Samples tab lists tests from your batch and displays the results

for each sample.

Figure 5-20 shows a Sample tab example listing three tests. Notice that the left pane displays Test IL-4, IL-6 and IL-8 with Test IL-4 selected. The right pane shows the IL-4 sample results. Notice that the well G3 location test result is “<10” and the associated Comments column for the third sample indicates a sample out of range error “Sample High/Low” because this sample has an MFI less than the lowest standard in the standard curve for this noncompetitive batch.

The first sample is within the standard curve range and thus displays an unflagged test result.

The system does not flag results as normal or abnormal according to a defined range. Error flags only note samples that fall below or above the standard curve.

The Comments column cells are editable and the information is displayed in reports.

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Replicate

Averaging

Figure 5-20. Analysis Window - Samples Tab Open

Standard and control replicates are predefined in the template. You define unknown sample replicates in batch setup indicated by replicate Sample ID.

The data analysis function supports replicate sampling. It calculates each sample as an individual sample, which is then averaged to obtain a replicate average.

• Standards tab—displays standard and control average values.

• Samples tab—displays sample average values.

Replicate averages are displayed with AVG in the Loc (location) column.

The AVG entry appears immediately afte r the we lls being averaged or at the end of the list depending on what you select under Sort (Sequentially or Alphabetically).

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If the system fails to determine the results for a replicate due to an excessive skew of one of the samples, you can invalidate the out of tolerance value. Use the Invalidate Standard (F4) or Invalidate Control (F6) buttons at the bottom of the Analysis window. Be aware that this fixes standards and controls, not patient sample. If any of the replicate standards and/or controls are invalidated, the “Avg” results reflects the average of the remaining standard and/or control replicates.

The system flags the failed sample so you may calculate the replicate set’s average without including the failed result in the equation.

You can sort the batch samples Sequentially (by the order in which they are acquired), or Alphabetically (by sample ID). Select the sorting method in the Sort group box.

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Commands This section describes the different commands used in the Luminex

IS 2.3 software. They are grouped Alphabetically

Acquire Patient This command, which is part of the Insert command/menu, adds

patients to the command list. After adding the patients you define the Sample ID and Dilution Factor for each new patient. The Dilution Factor defaults to 1.

Add Batch When creating a multibatch, adds an existing batch to a multi-batch. Alcohol Flush Removes air bubbles from the sample tubing and the cuvette using

70% isopropanol or 70% ethanol. The alcohol flush takes about five minutes. Uses the Luminex XYP reservoir due to volume requirement.

Autoscale Automatically adjusts the maximum number of events shown on the

Y axis on the histogram. Click during acquisition to readjust the Y axis scale.

Autosize Automatically adjusts column widths to fit the data and header sizes. Backflush Removes obstructions from the fluidics pathways by drawing sheath

fluid from the sheath fluid container. You do not need to supply solution in a plate. A backflush command takes about seven seconds.

CAL1 Runs the Classification portion of the Luminex System calibration. CAL2 Runs the Reporter portion of the Luminex System calibration. Cancel (Command) Cancels the process for the last command initiated. Cancel All Cancels all the commands in progress. Change Lot Opens the Change Lot dialog box from which you select a lot to

apply to a batch.

CON1 Runs the verification for the Classification calibration. CON2 Runs the verification for the Reporter calibration.

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Connect to Instrument Establishes a connection between the PC and the analyzer. Create New Multi-Batch This command opens the Luminex Multi-Batch dialog box, in which

you add or create new batches to add to a multi-batch.

Delete Batch Opens the Delete Batch dialog box, which shows a listing of the

unprocessed batches in the database. When you highlight a batch and click Select in this dialog box, the system deletes the selected batch.

Density/Decaying Toggles between the default density dot plot and the decaying dot

plot views.

Display Confirmation Screens

Disconnect from

Enables confirmation dialog boxes to display when you initiate many of the maintenance commands.

Disconnects communication between the PC and the analyzer

the Instrument Drain Used during troubleshooting to help remove debris from the bottom

of the cuvette. When draining, you do not need to supply solution. Draining takes approximately two minutes and should be followed by an alcohol flush with 70% isopropanol or 70% ethanol. Any fluid that drains from the system drains to the Luminex XYP reservoir as the default. However, you can set the system to drain to any unused well on the microtiter plate. The drain function normally expels 125

µL of fluid.

Eject/Retract If the XYP plate is retracted, this command ejects the microtiter

plate. If the XYP plate is already ejected, this command retracts the plate.

Enable Raw Data Storage Saves bead event data to the database. Export Batch Data Opens the Open Batch dialog box, from which you choose a batch to

export to an output.csv file.

Export CAL Opens a dialog box in which you choose the calibration file to export

to an output file.

Export CON Opens a dialog box in which you choose a control (validation) file to

export to an output file.

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Export Data Exports data to a .csv file in the batch folder. There is no dialog box

to confirm that the data was exported.

Help Opens the help file. Use this command if you need instructions on

using a feature in the Luminex IS 2.3 software.

Import Calibration Opens a dialog box in which you select a calibration file that

contains calibration lot information to import to use for calibrating the analyzer.

Import Control Opens a dialog box in which you select a control file that contains

control lot information to import for use when verifying calibration.

Import Template Opens up the Import Template dial og box, from which you select a

template to import for use in a batch.

Insert This combination menu/command on the Batch Setup window

allows you to add a user-defined number of patients to the command list, or skip a defined number of wells on the microtiter plate. The two options on the menu are Acquire Patient and Skip.

Invalidate Control Invalidates or removes a control from the data analysis. Luminex

does not recommend invalidating controls.

Invalidate Standard Removes a standard from the data curve. Use this command if doing

so improves the curve fit. Use caution when doing so because invalidating a standard will greatly affect the curve fit and subsequently the sample results.

Load Patient List Opens the Open Patient List File dialog box, from which you select a

patient list text file to append to a batch.

Log/Linear Toggles the x axis scale on the histogram or dot plot between

logarithmic and linear modes.

Maximize/Minimize Toggles between maximum and minimum histogram and dot plot

views.

Next Test This command is found in the Analysis window. Use this command

to view the next test in the batch.

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New Advanced Batch Assay developers use New Advanced Batch to acquire data without

having to build templates. This provides the developer with a faster and more convenient tool to develop assays. It does not store the results in the Luminex IS 2.3 database. It writes raw data to the output.csv file if Auto Export is selected, and writes run files if Enable Raw Storage is selected. This feature is the primary use of the Acquisition Detail tab. When you click on the New Advanced Batch button, the Options dialog box opens, where you enter information about the batch, define bead events for the session, and define commands for plate layout.

New Batch Opens a dialog box in which you select a template to create a new

batch in the Luminex Batch Setup window.

New CAL Targ. Opens the Update CAL Targets dialog box, in which you enter

information about the calibration microspheres you are using to calibrate the system.

New CON Targ. Opens the Update CON Targets dialog box, in which you enter

information about the control microspheres you are using to verify system calibration.

New Lot Opens the Open Template dialog box, from which you select a

template to which you want to add a lot or edit lot information.

Open Batch 1. The Open Batch command selected from the file menu or toolbar

opens the Open Batch Dialog box, from which you choose an existing batch to use for acquisition.

2. The Open Batch command on the Analysis window allows you to view data from a different batch.

Open Help Opens the online help file. Open Incomplete Batch Opens the Open Run dialog box, in which you select a batch to

recover.

Open Multi-Batch Opens the Open Multi-Batch dialog box, in which you select a multi-

batch to use for acquisition.

Pause Pauses the comma nd that the system is currently processing.

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Previous Test This command is found in the Analysis window. Use this command

to view the previously run test in the batch.

Prime Removes air from the system’s fluidic pathways by drawing

Luminex xMAP Sheath Fluid from the sheath fluid container. You do not need to supply solution in a plate. Priming takes approximately one minute. This command should also be used as part of the daily startup routine.

Print Batch Worklist Prints the status of each of the commands in the command list, as

well as the microtiter plate graphic.

Print Report Prints the data interpretation report including the standards, controls,

graph of standards, and Samples data.

Recalc Recalculates an affected test after a change is made to lot

information. There are two options: Manual recalculate and auto. If the auto checkbox is selected, the Recalc button (manually recalculate) is grayed out. In the auto selection mode, changes automatically trigger the system to reanalyze the affected test. If you are not using the auto mode, the ReCalc button is available after you make a change such as changing a formula or lot, or invalidating or validating a standard or control.

Replay Batch Replays batch sample acquisition. All batch commands are replayed

allowing for batch acquisition viewing exactly as in real-time. You cannot replay New Advanced Batches. When running a Replay Batch command, the original acquisition data is not altered. A new file is created. The most common use is for system demonstration and training.

Report Raw Fluorescence Enables the median fluorescence intensity (MFI) to appear on the

Analyte Report.

Resume Resumes the process that was paused. Sample Probe Down Raises and lowers the sample probe to adjust the probe height. This

adjustment is recommended when using different microtiter plates or when changing sample probes.

Sanitize Uses the Luminex XYP reservoir due to volume requirement. The

sanitize command performs a similar function as the alcohol flush

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command. However, the sanitize command uses 10% to 20% household bleach to decontaminate sample lines and the cuvette after biohazard contact.

Save and Load If selected, displays the run batch tab immediately after you set up

batch information and click Finish.

Save Only Allows you to save the batch information. User can open the batch

later to acquire data.

Self Diagnostics Performs a self-diagnostics to see if the Luminex analyzer and all

system operations are functioning correctly. Self-Diagnostics tests these functions:

• Flash memory test

• Microcontroller RAM test

• Nonvolatile memory test

• DSP program CRC test

• DSP capture test

• High voltage module

• Channel backgrounds test

• Pressurization test

• Actuator test

• Syringe pump test

• Backflush valve test

• Debubbler valve test

Show Bead After selecting an entry from the drop-down list, sets the histogram

to show events for only one beadset [bead set number], <all gated> events within the gate, or <all> events inside and outside the gate. Select <all> (default) before setting the gate.

Single Step Selecting this option on the toolbar pauses the system in between

each command or sample acquisition within a batch.

Skip [wells] Enables you to deliberately bypass specific wells during batch

acquisition.

Soak Prevents salt crystals from forming in the probe due to air exposure.

Soaking the probe replaces sheath fluid in the probe with water. You should perform the soak function at the end of each day. The system uses at least 250

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Start Analysis Opens the Open Batch dialog box, from which you select a batch to

analyze. Once you select the batch, the Analysis window opens.

Start (Plate) This command initiates data acquisition on New Advanced Batches

and New Batches using system templates. During acquisition, the system draws sample from the microtiter plate for processing.

Statistics This command enables you to display selected statistics for sample

data. There are eleven different statistics: %CV, Trimmed %CV, Count, Trimmed Count, Mean, Trimmed Mean, Median, Trimmed Peak, Peak, Trimmed Std Dev, and StdDev.

Test Sort Orders This command is in the Data Export Tab of the Options tab. You can

select Alphabetical by test name, Sequential by Region ID, or by Template Setup Order

• Alphabetical by Test Name - the tests are sorted by alphabetical order

• Sequential by Region ID - the tests are sorted numerically by bead ID#.

• Template Setup Order - the tests are sorted n the order set up in template, regardless of name or ID.

Validate Control Opens the Validate Control dialog box. In this box, you choose to

validate all control tests (click Yes), or only a single test (click No).

Validate Standard Opens the Validate Standard dialog box. In this box, you choose to

validate all control tests (click Yes), or only a single test (click No).

View Batch Data Click to open a previously acquired batch that is stored in the

database. You cannot view New Advanced Batches.

Warmup Warms up the system to prepare the optics prior to sample

acquisition. The system automatically begins warming up when you turn power on; however, you will need to use the Warmup command if the system is idle for four hours or longer.

Wash Washes fluidics line. Sends fluid (distilled water) through the fluidic

lines in the system. Pulls the fluid from a well or the reservoir and runs it completely through the system to the waste receptacle.

Zoom Zooms or enlarges a specific area on the histogram or dot plot

display. Click and drag right to left to adjust the graph’s range.

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Procedures This section explains how to perform functions in the software.

Using the Online Help Help files describe Luminex System features and capabilities. You

may find these topics by searching the contents of books and topics or by searching an alphabetic listing of the topics and books.

Topics provide information and details regarding software features, system capabilities, and any number of related material.

Books usually contain a group of topics. These topics are grouped together because they discuss similar or related topics.

Hyperlinks provide instant access to another topic or subtopic with related information to the linked subject or term.

 To open the system’s online help:

1. On the Help menu, click Contents.

2. In the Help dialog box, scroll through the contents and select the desired topic. Also, consider the index to locate information.

3. Double-click the help topic that you want to view. A topical dialog box opens with information on that topic.

 To display device information about the Luminex analyzer,

Luminex XYP instrument, and the Luminex LXR SDK:

On the Help menu, click About the Device. The resulting dialog box shows information that may be helpful when contacting Luminex Technical Support. Click OK to close the dialog box.

 To display information about the system software:

On the Help menu, click About the Software. The resulting dia- log box shows information about the system software. This includes software version, build number, and the system copyright information. Click OK to close the dialog box.

 To display system information:

1. On the Help menu, click About the Software. The resulting dialog box opens that displays software information.

2. Click System Info. The System Information dialog box opens. Click X in the top-right corner to close the dialog box.

3. Click OK to close the dialog box.

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Setting Software Options Set up and customize the system software and enter your company

information in the Options dialog box. The Software Options dialog box has three tabs: the General tab, the Company Information tab, and the Data Export tab.

 To configure software options:

1. On the Tools menu, click Options.

2. Click on the tab in which you want to set options. A full description of each tab and its options is listed below.

3. After you have entered and selected all of your preferences, click OK.

Define General

Tab Information

You define the following options on the General tab. Default Batch Directory. Names the default directory to store

batch information. Click the browse button and navigate to the desired folder (directory).

Current User. Enter the name of the current user or operator. The name appears on reports.

Analysis Display Digits. Use this feature to customize the number of digits shown on the Data Analysis dialog box and printed reports. The data is stored with its full precision (that is, including all digits), but the data appears as requested. The default analysis digit display is for two digits to show in the analysis.

Display Confirmation Screens. Enables confirmation dialog boxes to display when you initiate many maintenance commands. If you disable the confirmation screen display option, the confirmation screens remain for commands initiated from the Home tab.

Enable Raw Data Storage. Select this feature to save bead event data that is acquired while processing batches in the database. The system defaults to Enable Raw Data Storage. Raw data storage is necessary particularly when you use file mode (Replay Batch).

Report Raw Fluorescence. Select this feature to enable the median fluorescence intensity (MFI) display to appear on the Analyte Report. This feature was previously used to display MFI values on all reports. In the Luminex IS 2.3 software, the only report affected by this selection is the Analyte Report. All others are hard coded by the system.

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Auto-Start Analysis. Select this feature to enable the software to begin analyzing data immediately after processing batches. The automatic analysis feature takes place after the system finishes acquiring data while processing batches.

Define Company

Information Tab

Figure 5-21. Options Dialog Box—General Tab

You can enter information about your company and a company logo into the Company Information tab. This information is stored in the Registry for reference. The logo will not alter any Luminex IS 2.3 report or screen.

Company Information (name, address, phone and fax numbers, and location of company logo).

Use the browse button to navigate to the location where the logo is stored.

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Figure 5-22. Options Dialog Box—Company Information Tab

Define Data Export

Tab Information

Use the Data Export tab to configure your export data. See Figure 5-

23. The following options are available:

Auto Export Batches. Select this feature to automatically export the .csv file formats when the system finishes analyzing the batch. This allows you to run your own programs on exported data without having to manually start the export. This feature also takes place after acquisition completes. If this is not selected while running a New Advanced Batch, you must right-click on the data grid to get your output.csv file.

Copy Output.csv file to Common Output Dir. Select to send a copy of the Output.csv file to the My Batches/Output folder.

Prompt for Batch Comment. Check this button to initiate a prompt for batch commenting when a batch is finished.

Write Sample Comments. Select to add sample comments to the Notes column in the output.csv file.

Additional Export Stats. Select to define which sample statistics to export outside the Luminex IS 2.3 software to the Output.csv file.

Test Sort Order. Choose an option to define the sorting order. For more information on the options, see the Command section, beginning on page 5-31.

Additional Batch Information. Select one or more options to add additional information to the exported batch file (output.csv).

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Export Location Label Style. Choose one of these options to define the label data style exported to the Output.csv file. You can select sequential numbering, by plate location, or both (default).

Figure 5-23. Options Dialog Box—Data Export Tab

Setting up the Favorites List

 To add templates to the Favorites list:

1. On the Favorites list, click Add Template.

2. In the Open Template dialog box, double-click the template to

add to your Favorites list.

 To add commands to the Favorites list:

1. Click Add Commands from the Favorites list. The Command List dialog box opens.

Figure 5-24. Command List Dialog Box

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2. Select the command that you want to add to your Favorites list. For certain commands, you will need to select the location where you want to draw or expel fluid. Use the Location drop-down menu to open the microtiter plate image. Click on the microtiter plate location on the image. Ensure that the location you select is compatible with the volume intended for that location.

3. Click OK to add the command. The command displays in the Favorites list.

 To remove items from the Favorites list:

Select the item you want to remove from the Favorites list, then click Remove. The item disappears from the list.

Startup Procedures

Warm Up the

System

Warm up the system to prepare the optics prior to sample acquisition. The system automatically begins warming up when you turn power on. This procedure takes approximately thirty minutes.

Caution: Failure to properly warm up the system will effect assay results and system performance.

After four hours of inactivity , the status bar appears red and indicates that the lasers are off. You need to warm up the system again by manually initiating warmup.

 To warm up the system:

Click Warmup, then click OK. The command list on the Run Batch tab indicates that the system lasers are running. The Device Activity box on the Status Bar indicates that the system is warming. The Laser Status section on the Status Bar is yellow as it counts down from 1800 seconds. Upon completion, the Laser Status bar turns green and displays Warmed Up .

Prime the System Prime your system as part of the daily startup routine and as

necessary to remove air from the system’s fluidic pathways after:

• refilling the sheath container

• removing and replacing the sheath container

• observing air in the tubing

• changing the sheath fluid filter

• changing the syringe seal

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When priming, the system draws Luminex xMAP sheath fluid from the sheath fluid container and sends it directly out to waste, and takes approximately one minute. You do not supply solution in a plate.

 To prime the system:

Click Prime, then click OK to confirm that you want to prime the system. The status bar indicates that the prime command is processing.

Backflush the

System

Run an Alcohol

Flush

Backflush the system:

• t o remove obstructions from the cuvette

• if fluid does not flow through the waste tubing during prime cycles or during sample acquisition

• i f fluid drips from the sample prob e during priming and forms puddles of fluid on the plate

During a backflush, the system draws sheath fluid from the sheath fluid container and sends it directly to waste. You do not need to supply solution in a plate. A backflush takes between 10 and 30 seconds.

 To clear obstructions from the cuvette:

Click Backflush, then click OK to verify that you want to backflush the system. The status bar shows that the backflush command is processing.

Alcohol flush the system to remove air bubbles from the sample tubing and the cuvette using 70% isopropanol or 70% ethanol. The cuvette is the principal fluid pathway within the optics component of the system where the system reads the sample.

 To remove air bubbles from the sample tubing and cuvette:

1. On the Maintenance tab, click Eject/Retract

2. A confirmation dialog box opens telling you to place solution in the reservoir.

3. Put 70% isopropanol or 70% ethanol in the reservoir.

4. Click OK. The plate holder retracts, and the system performs the Wash command.

Run a Wash Cycle Use the wash cycle after an alcohol flush or as needed. For example,

wash four times with distilled water after calibration and twice with

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distilled water after Sanitize. Place at least 200 µL in a microtiter well or fill the Luminex XYP reservoir with distilled water. Washing takes about 30 seconds.

You should wash after calibration and verification, between batches and multi-batches, after sanitize, and before daily shutdown.

 To perform a Wash command:

1. On the Maintenance tab, click Eject/Retract.

2. Select Reservoir from the dropdown menu next to the Wash button, then click Wash. A confirmation dialog box opens telling you to place solution in the reservoir.

3. Put distilled water in the reservoir.

4. Click OK. The plate holder retracts, and the system performs the Wash command.

Set Luminex XYP

Instrument Heater

Temperature

Refer to your assay kit instructions to see if the assay needs to be analyzed at a particular temperature. If the instructions indicate that the Luminex XYP instrument heater is needed, set the heater to the specified heat setting. The user definable heater range is 35°C to 60°C. Use the heater only with the heater block in place.

Luminex recommends using a Costar® Thermowell® thin-wall polycarbonate 96-well plate (nonskirted), model P over the heater block sent with the Luminex System. Do not use standard 96-well microtiter plates if you are using the heater block.

Any temperature that you set remains in effect until you set another temperature or turn off the Luminex XYP instrument plate heater or exit the software.

The system displays the target temperature in the box below the T u rn ON button. Before the heater block temperature reaches the new temperature setting, the XYP Heater Temperature thermometer is red. Upon reaching the target temperature, the thermometer turns green. See page 5-9 for more information about the system monitor.

Warning: The heater plate of the Luminex XYP instrument is hot

when in use and may cause burns. Do not touch the heater plate.

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 To set the Luminex XYP instrument heater temperature:

1. Click Eject/Retract to eject the plate holder.

2. Set the Luminex XYP heater block on the plate holder.

3. Click Eject/Retract to retract the plate holder.

4. In the Temperature and Gauges area of the Run Batch tab, click Turn ON. The light on the button turns green and the thermometer fluid turns red as it raises to reach the target temperature.

5. Use the up and down arrows beneath the target temperature box to set the temperature you want the Luminex XYP instrument heater block to maintain. The user definable heater range is 35°C to 60°C.

6. Wait for the heater to reach your selected temperature and stabilize before processing samples. The thermometer turns green when the temperature is stabilized.

Calibration Procedures Calibrator xMAP microspheres are used to normalize the settings for

the reporter channel, both classification channels, and the doublet discriminator channel. Control xMAP microspheres are used to verify calibration and optical integrity for the system.

Calibrate the system at least once a month and:

• Following installation

• if the system is moved.

• if a part is replaced.

• if the delta calibration temperature shown on the system

monitor (on the Diagnostics tab) is more than ±3 degrees.

• if sample acquisition is problematic.

Each step in the calibration procedure usually takes less than one minute. You must run xMAP controls after each calibration. Once calibrated, the calibration values remain until you calibrate again. You can track system calibration and verification results through the Calibration Trend Report and the System Control Trend report. If you need target value information for Calibration or Control microspheres, you can find the information on the Luminex website at

http://www.luminexcorp.com. Click on the Support link then

navigate to the FAQ page on the Support page.

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Ensure that the Luminex analyzer lasers are warmed up and the probe height is set correctly before calibrating the system. Do not move the system waste line while calibrating.

You can run calibration and verification commands from the Maintenance tab. You can import and export calibration and control lots and reuse existing lot information for calibration and controls.

Run System xMAP

Calibrators

Note: When dispensing calibra-

tion and control microspheres, hold the bottle upside down at a 90-degree angle to the microtiter plate to ensure that you are getting accurate drop volume.

 To calibrate your system with xMAP calibrators:

1. Vortex the xMAP calibrator and control containers to ensure homogeneity. Do not dilute xMAP calibrator or control reagents.

2. Load a microtiter plate with at least five drops of each: CAL1 in well A1, CAL2 in well B1, CON1 in well C1, CON2 in well D1 and distilled water in well E1 through H1 to wash a total of four times. Use different wells as necessary. To select different well locations in the software, click on the drop-down arrow next to the entry cell for the calibrator or control, then click in the well location on the microtiter plate image.

3. Click Eject/Retract, then place the plate on the plate holder.

4. Fill the Luminex XYP reservoir with a solution of 70% isopropanol or 70% ethanol.

5. Click Eject/Retract.

6. On the Maintenance tab, click Prime. Click OK and wait for the Prime to finish (about 1½ minutes).

7. Click Alcohol Flush. Click OK and wait until the alcohol flush completes. The Device Status section in the status bar changes from yellow to green and displays “Standby”.

8. Click New CAL Targ. to enter or confirm calibration lot numbers in the Update CAL Targets dialog box. See Figure 5-

25.

9. Enter the CAL1 lot number and expiration date as printed on the bottle. Enter the values listed on the Certificates of Quality (COQ) included with your calibrators into the CAL1 boxes. If you are using a previously entered lot, select it from the dropdown menu in the Current CAL1 group box or click Import to import the information. See page 5-50 for more details on importing a Calibration lot.

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Figure 5-25. Update CAL Targets Dialog Box

10. Repeat step 9 for the CAL2 lot.

11. In the Maintenance tab, click CAL1, then click OK. The device

status section in the status bar changes from “Running” to “Standby”.

12. Click CAL2, then click OK. Wait until CAL2 completes.

The Device Status section in the status bar changes from “Running” to “Standby” and the Diagnostics tab turns red. The System Monitor on the Diagnostics tab displays the date and time in green if CAL1 and CAL2 are successful.

The Diagnostics tab turns red if all substances (CAL or CON) have not been run and under the following conditions:

• the first time the software is opened

• the first time a system is calibrated

• a new database is installed

• an old database is restored

• a CAL or CON fails.

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You must run system controls following calibration. Continue with the following “Run System xMAP Controls” section.

Run System xMAP

Controls

Run System xMAP controls to verify calibration. Dispense the Control microspheres into the microtiter plate at the same time that you dispense the Calibration microspheres. See “Run System xMAP Calibrators” on page 5-47.

 To verify system calibration with controls:

1. In the Maintenance tab, click New CON Targ. The Update CON Target Information dialog box opens. See Figure 5-26.

2. Enter the CON1 lot number, expiration date, and the values listed from the COQ included with your system controls into the CON1 entry boxes. If you are using a previously entered lot, select it from the drop-down menu in the Current CON 1 group box or click Import to import the information. See page 5-50 for more details on importing a lot.

3. Repeat step 2 for the CON2 lot information.

Figure 5-26. Update CON Targets Dialog Box

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4. Ensure that the analyzer is set to draw CON1 and CON2 beads from the wells you loaded in step 2 of “Run System xMAP Calibrators” on page 5-47.

5. In the Maintenance tab, click CON1 then click OK. Wait until CON1 completes. The Device Status section in the status bar changes from “Running” to “Standby”.

6. Click CON2, then click OK. Wait until CON2 completes. The Device Status section in the status bar changes from “Running” to “Standby”. The System Monitor on the Diagnostics tab displays date and time in green if both CON1 and CON2 are successful.

7. Ensure the analyzer is set to the draw distilled water from the well you loaded it in step 2 of “Run System xMAP Calibrators” on page 5-47.

8. Click Wash to wash the system after running the system calibrators. Wash a total of four times. You will need to change the well location. Click on the dropdown menu located to the right of the Wash button.

Selecting Existing

CAL or CON Lots

Importing CAL or

CON Lots

9. Click OK and wait until the wash completes. The device status section in the status bar changes from “Running” to “Standby”.

If an error occurs during system calibration or verification, an X appears in front of the Diagnostics tab title, and the title text turns red.

 To select an existing lot:

1. On the Maintenance tab, click New CAL Targ or New CON Targ.

2. Select a lot using the arrows located to the right of the Import and Export buttons in the Update CAL Targets and Update CON Targets dialog boxes.

Review the lot information and press OK to select the calibration lot.

 To import system CAL or CON lots:

1. On the Maintenance tab, click New CAL Targets or New CON Targets as appropriate. An Update CAL Targets or Update CON Targets dialog box opens.

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2. Click Import CAL or Import CON as appropriate. The Open

dialog box opens.

3. To select the calibration lot or control lot to import, click the

drop-down arrow for the Look in box. Browse for the appropriate folder, diskette, or CD location. The file type is a .lif file.

After you select the location, the available lots display in the selection list. Click the name of the lot to import and click Open. The lot name appears in the product information box. The lot and target information is displayed on the Update dialog box.

4. Click OK to complete the operation.

Exporting CAL or

CON lots

 To export system CAL or CON lots:

1. On the Maintenance tab, click New CAL Targets or New CON Targets. An Update CAL Targets or Update CON Targets

dialog box opens. Ensure you have the desired lot to export displayed or selected.

2. Click Export CAL or Export CON as appropriate.

3. In the Save As dialog box, select the folder (directory) where you want to export the lot as the Save-in location. The default is the Backup folder (directory) found in

Files\Luminex\Luminex 100 IS\Back up

4. Enter the lot name for the exported lot into the File Name box.

5. Click Save, then click OK. The dialog box closes. After you click OK, you can use the lot target values with the next calibration or verification.

The system saves the lot to the existing software as a lot accessible for the next calibration and/or verification. Once you export the desired calibration or control lot you can save it to disk to import to another computer.

C:\Program

Batch Setup Procedures A batch consists of a group of samples processed under control of a

template. Batches are set up using templates defined by assay kit manufacturers. Batches consist of templates and samples for acquisition, and can span more than one plate. Templates contain predefined commands that must be included in every batch acquisition.

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You can group batches together as a multi-batch. Multi-batches can consist of any number of batches that have been setup from different assay templates and are processed consecutively.

The assay kit manufacturer may provide templates in the kits, which they distribute on diskette or CD. Templates typically include assay standards, controls, and maintenance commands (such as washes or primes to acquire along with samples). OEM manufacturers may provide templates pre-installed with your system.

The kit manufacturer includes assay reagents in the assay kit. You must provide information about these reagents, such as lot numbers and concentration values for the standards and assay controls.

A batch can include samples across more than one plate. When setting up a batch, if the number of samples exceeds the wells in one microtiter plate, another plate appears for additional samples. The new plate appears to the immediate right of the existing plate image on the screen with a dark line between the adjacent columns of the two plates.

Importing

Templates

During acquisition, the Run Batch tab displays the wells containing the samples in the microtiter plate. Colors indicate the progress in analyzing the samples. The following well colors indicate wellacquisition states:

• Green well: with command number—sample not acquired.

• Yellow well: sample currently in acquisition

• Red well: sample failed. Check the system monitor for more information

• Green background: with check mark—successfully completed

Common sample errors are due to lower number of events acquired than established in the template.

You need to import new templates to the system only once. You must enter lot information for the standard and control reagents as specified in the template. This lot information is used for every batch setup using the template until it is changed.

Templates include standards, controls, both standards and controls, maintenance commands, and acquisition commands.

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 To import a template from a diskette or CD:

1. Insert the kit’s diskette or CD into the appropriate drive.

2. On the File menu, click Import Template. The Import Template dialog box opens.

3. Click the Look in drop-down arrow and navigate to the diskette or CD drive containing the template. The diskette drive is typically drive A and the CD drive is typically drive D.

4. The kit manufacturer’s template appears on the selection list. Click the name of the template.

5. Click Open to load the template.

Create a New

Batch

 To create a new batch:

1. Read the instructions provided with the assay kit you are using. Follow the kit instructions for any preparations.

2. Click New Batch. The Open Template dialog box opens.

3. Double-click the template you want to apply to the new batch. The template loads, and the Luminex Batch Setup window opens. See Figure 5-27.

Figure 5-27. Luminex Batch Setup Window

4. Enter the batch name (if different from the default name), a description (optional), and the creator’s name (optional).

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5. If you want to insert an Acquire Patient or Skip command, select

the command from the Insert menu. In the multiplier box, enter the number of patients that you want to add to the list or the number of wells that you want to skip and click Apply. Skipped wells and patient wells added to the batch are shown as green wells on the microtiter plate image.

6. If you are running a maintenance template, add any samples (for

processing). Then click Save and Load (default) or Save Only. Otherwise, continue with step 7.

7. If you want to change the well location where you begin acquiring samples, drag the highlighted starting well (default is A1) to the desired location on the microtiter plate.

8. Click the field in the Sample ID row that represents the last empty well on the microtiter plate.

9. Enter the sample ID for the sample to add. Repeat this step to add all of the additional samples to the batch. You can enter the sample manually, through a patient list, or using the system barcode reader. Batches may span more than one plate. When the first plate is full, a blue line separates the columns of the first plate with those of a second plate. To add a sample ID to the end of the list, press the Enter key or double-click in the last line.

Note: If any of the acquire sam-

ple commands within the template of the batch has an unassigned Sample ID, the system applies the first patient ID in the list to the unassigned sample acquisition command. The system appends any remaining patient IDs to the end of the command list in the order as they appear in the patient list.

10. To add a patient file to the batch, click Load Pa List. An Open Patient List File dialog box opens. See Figure 5-28. If you d o not want to add a patient list to the batch, skip to step 11.

Figure 5-28. Open Patient List File Dialog Box

11. Select a patient file to append to the batch and click Open. The system appends the patients to the batch.

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If all patient IDs in the batch are identified, the system appends the patient list items to the first empty location following the batch’s last command list activity. See “Add a Patient List” on page 5-62 for information regarding the patient file format.

12. Verify the dilution factor settings, and adjust as necessary. See page 5-65 for more information.

13. Select Save and Load (default) or Save Only.

14. Click Finish. If you selected Save and Load, the Run Batch tab opens displaying the batch, including the samples you added. If you selected Save Only, the system becomes idle as it waits for you to initiate a command.

15. If you selected Save and Load, load the plate using the Eject/ Retract button, then click Start Plate.

Open a Batch Use this procedure to open an existing batch. The batch name and

description appear on the Run Batch tab when you load the batch. You can open “Saved Only” batches with this procedure.

Copying and

Exporting Batch

Data

Clear a Batch from

the System

 To open a saved batch for an acquisition:

1. Click Open Batch. The Open Batch dialog box opens.

2. Double-click on the desired batch. The system loads the batch as you created it in the Run Batch tab.

3. Click Start Plate to initiate batch acquisition.

 To export Batch Data:

Right-click in the Batch Data area of the Acquisition Detail tab. In the right-click menu, click Copy to copy the currently displayed data to the clipboard. Click Export to manually export the currently loaded batch to the appropriate Output.csv file.

The Clear Batch command clears the entire batch from the Run Batch tab or the Message Log on the Diagnostic tab. Once you choose to clear the batch and verify that you want to continue with the command, you can recover th e cleared batch if it has not been run by clicking Open Batch.

 To clear a batch from the system:

1. Right-click on the area to clear.

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2. Click Clear in the dialog box.

3. Click Yes to confirm that you want to clear the batch.

Replay a Batch You can reprocess batches through the system multiple times using

Replay Batch. Replay Batch uses the data stored in the run files from the initial acquisition to reprocess a batch, creating a new batch output file.

Each time you reprocess a batch using Replay Batch, the system handles it as if it is a new batch; thus, creating a separate processed batch entry and output file. The initial batch data and output file always remain intact and unchanged.

You reprocess a batch using Replay Batch to:

• Run as demonstrations to see how the system processes samples and analyzes the results.

• Test a batch using different parameters, such as setting a different number of events to be collected or using a different bead map or new formula for analysis, also using a different template.The number of beads for collection must be less than or equal to the number of previously collected in the original sample.

If you reprocess a batch with the same template parameters in a different template, the system obtains results identical to the original batch. If you reprocess a batch using changed parameters, the system may obtain different results. When you replay a batch it labels unknown samples as Pa1, Pa2, Pa3, and so on. If you replay a batch containing replicates, replicate averaging will not be calculated in data analysis.

A number of variables can affect the final test results. You may also change the standards or controls processed with the batch or multibatch. These variables may effect your test results:

• minimum number of events for acquisition

• f ormula used to analyze the MFI values

• standards or controls validation or invalidation

• t ype of analysis (qualitative, quantitative, acquisition only, or maintenance)

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 To reprocess samples using Replay Batch:

1. On the Acq. Detail tab, click Replay Batch. The Browse for Folder dialog box opens displaying the My Batches folder.

2. Select the desired batch under the My Batches folder and click OK.

3. The Open Template dialog box opens. Click on the desired template and click Select.

4. The Run Batch tab becomes the active tab. You can monitor the commands as they process. Click on the Acq. Detail tab and monitor the data, histogram, and dot plot.

After replaying a batch, you can analyze the data.

 To analyze Replay Batch data:

Note: The Open Batch

dialog box does not list or show the templates associated to the batches.

1. Click Start Analysis.

2. In the Open Batch dialog box, select the batch you want to analyze, then click Select. The most recent Replay Batch is the last or has the highest ID number. See Figure 5-29.

Figure 5-29. Analyze Open Batch

The Analysis window opens, showing the Batch info in the Standards tab. To close the Analysis window click Close. See “Analyzing Batches and Multi-Batches” on page 76..

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Delete a Batch You can only delete unprocessed batches. Batches are deleted from

the Open Batch list and moved to the Open Incomplete Batch list.

 To delete an unprocessed batch:

1. On the File menu, click Delete Batch. A dialog box opens listing

the unprocessed batches in the database.

2. Select the unprocessed batch that you want to delete.

3. Click Select to delete the batch.

If you need to recover a deleted batch, select Open Incomplete Batch from the File menu.

Create a Multi-

Batch

A multi-batch is a set of batches that you want to process consecutively. You can add batches to the multi-batch from existing batches in your database, or you can create new batches to add to the multi-batch. You can include as many batches as you need. The software does not limit you to a certain number of batches per multibatch. Different batches within the multibatch are separated by thick red lines above the first well and below the last well of each batch. Multi-batches may span more than one plate. A blue line separates the graphical representation of the plates. A scroll bar appears along the bottom of the microtiter plate image so you can view additional plates.

 To create a multi-batch with new batches:

1. Click New Multi-Batch. The Luminex Multi-Batch Setup dialog

box opens, shown in Figure 5-30.

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Specifications and Main Features

Frequently Asked Questions

User Manual

Contents

Introduction

About This Manual This manual provides you with information to understand and use the

Intended Use The Luminex 100 IS 2.3 system is designed to perform a wide range

Technical Support Luminex Technical Support representatives are ready to help you,

Luminex Website Additional information is available on the Luminex website. Search

Safety

Symbols Please become familiar with the information in this chapter before

Warnings and Notes Informational notes and warnings appear in this manual.

Safety Precautions Read the following safety information before setting up or using the

FCC Label This equipment has been tested and found to comply with the limits

Fluidics This system contains fluidics. In the event of a fluid leak, turn off all

Luminex 100 Analyzer Laser

Barcode Reader Laser This label is attached to the barcode reader.

Mechanical

Biological

Heat

Blue Indicator Light

Decontaminating the Luminex 100 Analyzer for Return Shipment

The System

Theory of Operation Luminex 100 IS technology is based on flow cell fluorometry with

Hardware The Luminex 100 IS system includes the following hardware:

xMAP Technology Reagents

Required Laboratory Reagents

Luminex 100 IS 2.3 Software

Luminex 100 IS Performance Specification

Speed • USB communications link for fast data transfer

Accuracy and Precision • Sample uptake volume: ± 5%

Sensitivity • Detect 1000 fluorochromes phycoerythrin (PE) per xMAP

Capacity The specifications below reflect minimum capacity values:

Luminex 100 Analyzer General

Optics • Reporter laser: 532 nm, nominal output 10-15 mW, maximum

Fluidics • Sheath flow rate 90 µL ± 5 µL/second

Electronics • Reporter channel detection: A/D resolution 14 bits

Luminex XYP Instrument General

Luminex SD System General

PC Specifications For systems using a PC, A Dell OptiPlex GX280 or Dell Optiplex

Recommended Additional Equipment

System Overview The system consists of three subsystems: electronic, fluidic, and

Electronics

Fluidics

Optical The optical system consists of the optical assembly and the excitation

xMAP Technology Reagents

Basic Concepts

Background Information

Fluidics There are two fluidic paths in the Luminex 100 analyzer. The first

Excitation The excitation system in the Luminex 100 analyzer involves two

xMAP Microspheres The xMAP microspheres are highly uniform, polystyrene particles

Software Overview This section provides a brief overview for using the Luminex 100 IS

Using Luminex 100 IS 2.3 Software

Main Window The Luminex IS 2.3 software starts automatically when you log into

Menu Bar The menu bar contains the following menus: File Menu, Tools

Toolbar The shortcut buttons on the toolbar can be found in other locations in

Tabs This section describes the tabs in the Main window.

Status Bar The Status Bar displays information about the Command State,

Secondary Windows

Batch Setup Window In the Batch Setup window, you define information used to create

Analysis Window When the system analyzes batches, it displays the data in a three-tab

Commands This section describes the different commands used in the Luminex

Acquire Patient This command, which is part of the Insert command/menu, adds

Add Batch When creating a multibatch, adds an existing batch to a multi-batch. Alcohol Flush Removes air bubbles from the sample tubing and the cuvette using

Autoscale Automatically adjusts the maximum number of events shown on the

Autosize Automatically adjusts column widths to fit the data and header sizes. Backflush Removes obstructions from the fluidics pathways by drawing sheath

CON1 Runs the verification for the Classification calibration. CON2 Runs the verification for the Reporter calibration.

Connect to Instrument Establishes a connection between the PC and the analyzer. Create New Multi-Batch This command opens the Luminex Multi-Batch dialog box, in which

Delete Batch Opens the Delete Batch dialog box, which shows a listing of the

Density/Decaying Toggles between the default density dot plot and the decaying dot

Display Confirmation Screens

Eject/Retract If the XYP plate is retracted, this command ejects the microtiter

Enable Raw Data Storage Saves bead event data to the database. Export Batch Data Opens the Open Batch dialog box, from which you choose a batch to

Export CAL Opens a dialog box in which you choose the calibration file to export

Export CON Opens a dialog box in which you choose a control (validation) file to

Export Data Exports data to a .csv file in the batch folder. There is no dialog box

Help Opens the help file. Use this command if you need instructions on

Import Calibration Opens a dialog box in which you select a calibration file that

Import Control Opens a dialog box in which you select a control file that contains