Luminex 200 User Manual

Lum/nex®

Luminex 200™ User Manual for LDS Version 1.7

© LUMINEX CORPORATION, 1998 - 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.

Voice: [phone]

Fax: [phone]

Technical Support U.S. and Canada: toll-free 1-877-785-2323

Technical Support international: [phone]

Technical Support fax: [phone]

Luminex® 200™ User Manual for LDS Version 1.7

CN-M078-01

PN 89-00002-00-150 Rev. A

October, 2005

Luminex reserves the right to modify its products 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.

The following are trademarks of Luminex Corporation: Luminex, Luminex 100, Luminex HTS,

Luminex 100 IS, Luminex 200, LabMAP, LumAvidin, Luminex XYP, Luminex SD, xMAP, and FlexMAP. All

other trademarks, including Windows, Cheminert, Tween, Pentium, Dell, Alexa and Titertube are registered trademarks of their respective companies.

The contents of this manual and the associated Luminex software are the property of LUMINEX CORPORATION and are copyrighted. Any reproduction in whole or in part is strictly prohibited.

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 SOFTWARE 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 SOFTWARE or the sale 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 microspheres 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 TERMINATION. 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 patent, copyright, trade secret, trade name, 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. You 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 SOFTWARE PRODUCT IS AT 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 WARRANTIES OF MERCHANTABILITY, 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 INFORMATION, 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

Contents

Introduction

1-1

Principle of Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-1 Basic Concepts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-2 Fluidics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-2 Excitation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-3 xMAP Microspheres . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-3 Reporter Fluorochromes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-3 Fluorescence Compensation . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-4 Repetitive Microsphere Measurements . . . . . . . . . . . . . . . . . . 1-4 About this Manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-4 Technical Assistance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-5

Safety

2-1

Intended Use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-1

Safety and Regulatory Symbols . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-2

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

Safety Precautions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-3

Electrical . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-3

Fluidics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-5

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

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

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

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

Equipment Disposal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-8

System Overview

3-1

Hardware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-1

xMAP Reagents. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-1

Required Laboratory Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-2

Software. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-2

Luminex 200 Performance Specifications . . . . . . . . . . . . . . . . . . . 3-2

Speed. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-2

Accuracy and Precision . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-2

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

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

Luminex 200 Analyzer Specifications . . . . . . . . . . . . . . . . . . . . . . 3-3

Optics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-4

Fluidics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-4

Electronics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-4

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xMAP Technology

Luminex XYP Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-4

Luminex SD System General . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-5

PC Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-5

Recommended Additional Equipment . . . . . . . . . . . . . . . . . . . . . . 3-5

Printer. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-5

Uninterruptible power supply (UPS) . . . . . . . . . . . . . . . . . . . . 3-5

Surge Protector. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-6

Vortex . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-6

Bath Sonicator . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-6

Luminex System Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-6

Electronic . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-6

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

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

Luminex XYP Reservoir . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-12

Luminex XYP Heater Block . . . . . . . . . . . . . . . . . . . . . . . . . 3-12

Installation

4-1

Luminex 200 System Installation. . . . . . . . . . . . . . . . . . . . . . . . . . 4-1

Luminex 200 Software Setup . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-3

Adjusting the Sample Probe Height . . . . . . . . . . . . . . . . . . . . . . . . 4-4

Installing Software and Firmware . . . . . . . . . . . . . . . . . . . . . . . . . 4-6

What to Do Next. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-6

System Startup

5-1

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

Calibration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-2

Add New Classification Channel Calibration Lots . . . . . . . . . 5-2

Add New Reporter Channel Calibration Lots . . . . . . . . . . . . . 5-4

Calibrate the Luminex Analyzer . . . . . . . . . . . . . . . . . . . . . . . 5-5

Preview or Print Calibration Report . . . . . . . . . . . . . . . . . . . . 5-7

Analyzer menu commands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-9

Data Acquisition

6-1

Options . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-1

User tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-1

Acquisition tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-3

Advanced tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-3

Info tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-4

Setup XY Tab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-4

Sessions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-5

Create a Session Using Default Settings. . . . . . . . . . . . . . . . . 6-6

Create a Session Using a Template . . . . . . . . . . . . . . . . . . . . . 6-6

Open and Add to an Existing Session . . . . . . . . . . . . . . . . . . . 6-8

Open an Existing Session and Save it as a New Session. . . . . 6-9

Templates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-9

Create a Template . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-9

Modify a Template. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-9

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Contents

Delete a Template . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-10

Multiplex Acquisition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-10

Menu Bar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-10

Toolbar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-12

Main Control and Session Detail . . . . . . . . . . . . . . . . . . . . . . 6-12

Right-Click Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-14

Session Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-14

Histogram . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-17

Gates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-18

Analyzer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-19

Bar Graph/Dot Plot Display . . . . . . . . . . . . . . . . . . . . . . . . . . 6-20

System Monitor. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-22

Status Bar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-24

Data Collection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-24

Run an Entire Plate of Samples . . . . . . . . . . . . . . . . . . . . . . . 6-25

Run a Range of Samples. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-26

Run a Single Sample. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-27

Interrupt a Run . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-27

Reports. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-28

Maintenance and Cleaning

7-1

Startup . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-2

Shutdown. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-2

Check Fluid Levels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-3

Check for Leaks. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-3

Calibrate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-3

Sanitize . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-3

Clean Accessible Surfaces. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-4

Clean Sample Probe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-4

Replace Air Intake Filters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-5

Replace Syringe Seal. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-6

Clean Luminex Analyzer Ventilation Filter . . . . . . . . . . . . . . . . . . 7-7

Change Sheath Filter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-7

Change Sheath Quick-disconnect O-rings . . . . . . . . . . . . . . . . . . . 7-8

Change Fuses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-9

Adjust Sample Probe Vertical Height . . . . . . . . . . . . . . . . . . . . . . . 7-9

Storing the Analyzer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-10

Decontaminating the Luminex Analyzer for Return Shipment. . . 7-11

Troubleshooting

8-1

Power Supply Problems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-2

Communication . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-2

Pressurization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-3

Fluid Leaks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-4

Sample Probe. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-5

Calibration Problems. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-7

Acquisition Problems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-9

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Bead Detail Irregularities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-11

Error Messages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-13

Printing Errors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-14

Verification. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-14

Protocols

9-1

xMAP Microsphere Handling . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-1 Microsphere Dispersion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-1 Probe Sonicator . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-2 Bath Sonicator . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-2 Enumeration of xMAP Microsphere Suspensions. . . . . . . . . . 9-3 xMAP Microsphere Separation Methods. . . . . . . . . . . . . . . . . 9-3 xMAP Microsphere Agitation During Assay . . . . . . . . . . . . . 9-3 xMAP Microsphere Stability and Storage . . . . . . . . . . . . . . . . 9-4

Two-Step Carbodiimide Coupling of Protein to xMAP

Carboxylated Microspheres . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-4 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-4 Equipment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-4 Materials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-4 Preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-5 Procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-5 Technical Notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-6

One-Step Carbodiimide Coupling of

Oligonucleotides to xMAP Carboxylated Microspheres . . . . . . . . 9-7 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-7 Equipment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-7 Materials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-7 Preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-8 Procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-8 Technical notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-9

Binding Biotin-conjugated Molecules to

LumAvidin-Modified xMAP Microspheres. . . . . . . . . . . . . . . . . . 9-9 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-9 Equipment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-9 Materials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-10 Preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-10 Procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-10 Technical notes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-11

Glossary	A-1
Product Numbers	B-1

Hardware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B-1

Software . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B-2

Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B-2

Training . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B-3

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Luminex Sheath Delivery System

C-1

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

Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-1

General . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-1

Power . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-1

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

Hardware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-2

Sheath Delivery System Setup . . . . . . . . . . . . . . . . . . . . . . . . . . . C-3

Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-3

Using the Sheath Delivery System . . . . . . . . . . . . . . . . . . . . . . . . C-5

Audible Alarm and Fault LED . . . . . . . . . . . . . . . . . . . . . . . . C-5

Maintenance and Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . C-6

Replacing the Sheath Delivery System with Sheath Bottles . C-6

Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-6

Contact Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-7

Labels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-8

Index

Index-1

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vi	PN 89-00002-00-150 Rev. A

1 Introduction

Principle of

Operation

The Luminex® 200™ system is a compact analysis unit consisting of a Luminex analyzer, the Luminex XY Platform (Luminex® XYP™), a PC, Luminex software, and reagents. The Luminex analyzer integrates optics, fluidics, and advanced signal processing in a 17 inch by 20 inch footprint. The Luminex XYP provides efficient handling of 96-well microtiter plates. The PC uses Luminex software to control the analyzer. The optional Luminex Sheath Delivery system (Luminex® SD™) is used to provide sheath fluid directly from a 20 liter container.

Photodiodes and a photomultiplier tube receive fluorescent signals from xMAP® microspheres. The analyzer digitizes the waveforms and delivers the signals to a digital signal processor (DSP).

Proprietary algorithms function with the DSP to greatly increase the sensitivity.

Hardware and preloaded software simplify installation and setup, and with the help of this manual, you will be able to begin using xMAP technology.

The Luminex 200 system analyzes immunoassays, complex genetic analyses, and enzymatic assays in one simple format. The reactants (antibodies, oligonucleotides, substrates, etc.) of these assays are anchored to the surfaces of uniquely fluorescent microscopic beads, called microspheres. The unique fluorescent emission spectra of a given microsphere identifies each of the assays performed simultaneously on a single sample.

The Luminex analyzer individually analyzes xMAP microspheres in a flow stream. Every xMAP microsphere is accurately classified to

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Basic Concepts

Fluidics

its own subset, based on its fluorescent signature. In addition, the Luminex analyzer scans each microsphere for the presence of a reporter fluorescence that quantifies the assay at the microsphere’s surface.

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

Microtiter plates with 96 wells must be compatible with the Luminex XYP instrument heater block temperature when performing heated assays. The heater block temperature ranges from 35°C to 55°C (95°F to 131°F) .

The system reads the wells in column-first order. An entire column of wells is read; that is, well A1, B1, C1, D1, and so on to the end of the column. Then the XY Platform moves over to the next column and reads well A2, B2, C2, D2, and so on. When placing a plate on the plate holder, place it so well A1 is in the upper-left corner of the holder. You can run an entire plate, a consecutive range of samples from the plate, or a single sample from the plate.

Luminex has pioneered a versatile and robust technology for the measurement of soluble analytes. The Luminex 200 system performs simultaneous, discrete measurements of multiple microsphere-based reactions from a single specimen aliquot. A clear understanding of the concepts and functionality associated with the instrument and xMAP microspheres contributes to greater success with this technology. We present a brief overview of some of the basic concepts. For more information, refer to Practical Flow Cytometry, 4th edition, by Howard M. Shapiro, M.D. (New York: Wiley-Liss Inc., 2003).

There are two fluidic paths in the Luminex analyzer. The first path is 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 userspecified volume of sample from a microtiter plate to the cuvette. The sample is injected into the cuvette at a steady rate for analysis. After analysis, the sample path is purged with sheath fluid by the second fluidics path. This process expels sheath fluid into the sample

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Introduction

Excitation

xMAP Microspheres

Reporter Fluorochromes

container and effectively removes residual sample within the tubing, valves, and probe. The second fluidics path is driven under positive air pressure and supplies sheath fluid to the cuvette.

The excitation system in the Luminex analyzer uses two solid-state lasers. A reporter laser excites fluorescent molecules bound to biological reactants at the microsphere surface, and a classification laser excites fluorochromes embedded in the microsphere. The lasers illuminate the xMAP microspheres as they flow single-file through the cuvette. These fluorescent signals are discriminated with selective emission filters and are converted into intensity units by a digital signal processor.

The xMAP microspheres are highly uniform, polystyrene particles that have been crosslinked during polymerization for great physical and thermal stability. Varying amounts of fluorochromes embedded within each microsphere give each microsphere set an unique fluorescent signal. To ensure the stability of this address, it is essential to protect the xMAP microspheres from light. Do not subject xMAP microspheres to prolonged high temperatures, and protect them from freeze/thaw manipulations.

Each xMAP microsphere is dyed to emit light in a certain classification channel. All xMAP microspheres of a given emission represent a distinct assay within a multiplex of assays. A reporter channel is used to detect fluorescence bound to the surface of each microsphere, and each reporter emission quantitates each of the distinct assays. Only one reporter emission is used for multi-analyte assays in the Luminex analyzer. The Luminex analyzer uses an excitation wavelength of 532 nm, and has an emission wavelength of 575 nm ± 12 nm.

Fluorochromes R-Phycoerythrin and Alexa® 532 work well with the Luminex analyzer, but you are not limited to these two. You may have success using other fluorochromes such as Cyanine 3.

	R-	Alexa
	Phycoerythrin	532
Formula weight (Daltons)	240,000	470
Absorbance max (nm)	480, 546, 565	531
Extinction max (M-1cm-1)	1,960,000	83,800
Emission max (nm)	578	554

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Fluorescence

Compensation

Repetitive Microsphere

Measurements

About this Manual

Quantum yield	0.82	0.8

Fluorescent spillover occurs when an emission spectrum overlaps other emission criteria; therefore, selective filtering cannot occur. In most analyzers, emission spillover is corrected using a technique called compensation. Compensation involves subtraction of emission percentage from another emission signal. In the Luminex 200 system, compensation is not necessary—the reporter signal does not spill over into classification emissions.

Bound to reagents at the surfaces of the xMAP microspheres, the reporter provides raw analytical data. Because a microsphere suspension provides near liquid phase reaction kinetics, each microsphere theoretically binds an equal number of reporter molecules. Equal binding results in a statistically even distribution of reporter on each microsphere in a set. When the Luminex analyzer registers numerous events for each set, the system reports individual population values based on an even coating of reporter. This means that numerous replicates for each microsphere population are measured from a single reaction vessel. The confidence in a given measurement strengthens with increased replicate measurements. For adequate confidence, 100 collected events per microsphere set in each reaction vessel is usually sufficient. Measurements are reported as the median for the xMAP microspheres having a specific color signal.

This manual introduces you to the Luminex 200 system. With step- by-step instructions, the manual guides you through initial setup and powering-on procedures. Next, it offers a concepts section which describes some of the key topics encountered during system operation. Once you have a working knowledge of the analyzer, the manual’s text and figures lead you through the data collection process, giving detailed explanations of available options and offering examples when necessary. Glossary and troubleshooting sections assist as references. In short, this manual instructs you through each operational stage—from unpacking the Luminex analyzer and XYP to collecting your data.

The conventions in this document assume a basic familiarity with

computers and a knowledge of Windows® software. We typically document only one method of accessing a command, although many commands are available through more than one method, such as

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Introduction

Technical

Assistance

from the main menu bar, from the toolbar, and from menus that appear when you right-click an area of the screen.

You can find answers to frequently asked questions (FAQs) on our website: http://www.luminexcorp.com. From the Support menu at the top of the page, select FAQs.

For assistance, contact Luminex Technical Support via email. Users may reach us at [e-mail] or [e-mail].

Users in the U.S. and Canada can call us at 1-877-785-BEAD (1- 877-785-2323) between the hours of 7:00 a.m. and 7:00 p.m. Central Time, Monday through Friday. Users outside of the U.S. and Canada can call us at [phone] between the hours of 7:00 a.m. and 7:00 p.m. Central Time, Monday through Friday.

Users in Europe by can call us at [phone] between the hours of 8:30 and 5:30, Central European Time, Monday through Friday.

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2 Safety

Intended Use

Your safety is important. Please do not perform procedures on your Luminex 200 system that are not specifically contained in this manual, unless you are directed to do so by Luminex Technical Support. This chapter discusses the following safety topics

•Intended Use

•Safety and Regulatory Symbols

•Warnings and Notes

•Safety Precautions

•Equipment Disposal

The Luminex analyzer is designed for a wide range of indoor laboratory testing applications measuring biomolecular reactions on the surfaces of xMAP microspheres. The Luminex 200 for LDS Version 1.7 system is intended for general laboratory use.

The Luminex XYP instrument is intended for use with the Luminex analyzer. With the Luminex XYP, you can process 96-well microtiter plates without handling individual samples. The Luminex XYP works with microtiter plates that are no thicker than 0.75 in.

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Safety and

Regulatory Symbols

These symbols describe warnings, cautions, and general information used in the operation of this analyzer.

Symbol	Description	Symbol	Description	Symbol	Description
			Warning		Warning
	Alternating current		(refer to manual)		(refer to manual)
	(ac)
			Warning		Warning
	Protective ground		(refer to manual)		(refer to manual)
			Warning		Warning (refer to
	On		(refer to manual)		manual)
			Catalog Number		Consult instructions
	Off				for use
	WEEE - Do not throw		Batch Code		Serial Number
	in trash
	Temperature		Expiration Date		Date of Manufacture
	Limitation
	Manufacturer		Underwriter’s		Canadian
		US	Laboratory		Underwriter’s
					Laboratory
	European Union
	Conformity
Warnings and Notes		Informational notes and warnings may 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.

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Safety

Safety Precautions

Electrical

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

Please read the following safety information before setting up or using the Luminex 200 system. A user should always be present during system operation. This system contains electrical, mechanical, and laser components which, if handled improperly, are potentially harmful. In addition, biological hazards may be present during Luminex analyzer operation. Therefore, Luminex Corporation recommends that all Luminex analyzer 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 equipment is used in a manner not specified by Luminex Corporation.

The Luminex analyzer and Luminex XYP must be connected to approved power sources.

Do not perform any maintenance or cleaning of the system’s electrical components (except for the fuses).

This caution label appears on the back of Luminex analyzers and on the Luminex XYP:

Figure 2-1. Fuse Replacement Warning Label

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

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Figure 2-2. European Safety Requirements Label

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

The following voltage labels displays on the back of the Luminex analyzer:

Figure 2-3. Luminex 200 Voltage Label (Coherent)

This voltage label displays on the back of the Luminex XYP:

Figure 2-4. Luminex XYP Voltage Label

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Safety

Fluidics

Warning: If the system is used to test biological samples, use your standard laboratory safety practices when handling system waste.

The Luminex analyzer 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.

You must manually monitor waste levels. Do not allow the waste container to overflow. You can avoid this by emptying the waste container each time you fill the sheath fluid container. Do not place the waste container on top of the analyzer.

Be careful not to touch the seal in the waste bottle cap or allow it to become wet or dirty. This could cause the bottle to become pressurized, causing pressure errors in the system. If the seal gets wet, let it air-dry. Touching the seal can contaminate it. If it gets dirty, you should replace the waste bottle cap.

To maintain a stable flow rate, do not move the waste container or waste line during system operation. While you can move the line along a horizontal surface, do not permanently reroute the waste line elevation without first contacting Technical Support. You can move the line temporarily for cleaning and maintenance purposes.

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

This label appears on the back panel of the analyzer:

Figure 2-5. Laser Class Label

This label appears on the back panel of the Luminex analyzer:

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Figure 2-6. Laser Caution Label

Do not remove the analyzer cover. When performing routine maintenance, ensure that power to the analyzer is OFF and the power cord is disconnected.

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

Figure 2-7. Laser Caution Label on Optics Cover

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

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Safety

Mechanical

Note: Access doors must be closed while operating the instrument. An operator must be present during operation of the instrument.

Biological

Figure 2-8. Avoid Exposure Label

Caution: Use of controls or adjustments or performance of procedures other than those specified herein may result in hazardous radiation exposure.

Caution: To avoid exposure to hazardous radiation, only perform procedures and adjustments as specified in this manual.

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 sample probe. Keep hands and fingers out of the Luminex XYP slot during operation.

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 syringe arm.

Warning: Human/animal samples may contain biohazardous infectious agents. Where exposure (including aerosol) to potentially biohazardous materials exists, follow appropriate biosafety procedures and use personal protective equipment, such as gloves, gowns, laboratory coats, face shields or masks, 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 may be hot and could cause personal injury if touched. Do not touch the heater plate.

Blue Indicator Light

Equipment Disposal

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

The Waste Electrical and Electronic Equipment Directive 2002/96/ EC requires that you properly dispose of electrical and electronic equipment when it reaches its end of life.

NOTE: This directive applies to customers in the European Union (EU) only. Users outside of the EU may disregard this section.

If you are disposing of a Luminex 200 Analyzer, SDTM System, or XYPTM Instrument, you must decontaminate the system. Refer to “Decontaminating the Luminex Analyzer for Return Shipment” on page 7-11. Next, call Luminex Technical Support for a Recycled Returned Materials Authorization (RMA) number at [phone] between the hours of 7:00 a.m. to 7:00 p.m. Central Time, Monday through Friday. You will then return the equipment to:

Luminex Corporation

12201 Technology Blvd., Suite 130 Austin, TX 78727

For information on disposing of the PC or monitor, refer to manufacturer documentation.

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3 System Overview

Hardware

xMAP Reagents

The Luminex 200 for LDS Version 1.7 system includes the following hardware:

•Luminex analyzer

•Computer (PC), monitor, and accessories

•Luminex XYP instrument (with long sample probe)

•Power cables

•Short sample probe

•Reservoir

•Shield

•Heater block

•Sheath fluid container

•Waste container

•Sheath fluid line

•Air line

•Sheath fluid intake line

•Serial cable

•USB cable

•CANBUS cable

•Sample probe height alignment kit

In addition, you can purchase the optional Luminex Sheath Delivery system (Luminex SD).

•xMAP classification calibration microspheres (CAL1)

•xMAP reporter calibration microspheres (CAL2)

•Luminex xMAP Sheath Fluid

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Required

Laboratory

Reagents

Software

Luminex 200

Performance

Specifications

Speed

Accuracy and Precision

The following items are necessary to run the Luminex 200 system. They are not provided by Luminex.

•Household bleach

•70% isopropanol or 70% ethanol

•Mild detergent

•Distilled water

Luminex Data Collector software provides complete control of the Luminex analyzer and performs real-time digital analysis of assays. The Luminex 200 system is preloaded with the Luminex software. However, we supply a software CD in case 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.

•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

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

•Up to 100 xMAP microsphere sets per sample

•System warmup: 30 minutes. Systems that remain inactive for at least four hours will require a warm-up to restart the lasers. After acquiring sample, running system calibrators, running system controls, and warming up the instrument, the system resets the four-hour internal clock.

•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.

•Internal sample carry over: < 0.9%

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

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PN 89-00002-00-150 Rev. A

xMAP Technology

System Overview

Sensitivity

Capacity

Luminex 200

Analyzer

Specifications

•Detect 1000 fluorochromes phycoerythrin (PE) per xMAP microsphere

•Reporter channel dynamic range: 3.5 decades of detection

The specifications below reflect minimum capacity values:

•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

•Sample 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

•Microtiter 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: any color of flatbottom, conical, round, filter bottom, or half-well plates. The overall plate must not have an overall height of more than 0.75” (19 mm).

•Microtiter plates with 96 wells must be compatible with Luminex XYP instrument heater block temperature from 35°C to 55°C (95°F to 131°F) when performing heated assays and using the heater block.

•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.

PN 89-00002-00-150 Rev. A

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Luminex 200 User Manual for LDS Version 1.7

xMAP Technology

Optics

Fluidics

Electronics

Luminex XYP

Specifications

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

•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

•Sheath flow rate 90 µL ± 5 µL/second

•Cuvette: 200 micron square flow channel

•Sample injection rate: 1 µL/second ± .05 µL/second

•Sample uptake volume: 20-200 µL

• Reporter channel detection: A/D resolution 14 bits

•Communications interface: USB

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

•Humidity: 20% to 80%, noncondensing

•Altitude: designed to operate at 2400 m (7874 ft.) above mean sea level or below.

•Physical dimensions: 44 cm (17.25 in) W x 61 cm (24 in) D x 8 cm (3 in) H

•Weight: 15 kg (33 lbs)

•UL installation category: UL Installation Category II, as defined in Annex J of UL 3101-1.

•Pollution degree: UL Pollution Degree 2, as defined in section 3.7.3.2 of UL 6101A-1

•Heater operating range: 35°C to 60°C (95°F to 140°F) with tolerance 0°C to +2°C

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

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

•Communications interface: RS 232

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

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PN 89-00002-00-150 Rev. A