Beckman Coulter Cell Lab Quanta SC Instructions For Use Manual

Cell Lab Quanta SC

Instructions for Use

PN 721742AD (October 2011)

Manufactured for
Beckman Coulter, Inc.
250 S. Kraemer Blvd.
Brea, CA 92821

WARNINGS AND PRECAUTIONS

READ ALL PRODUCT MANUALS AND CONSULT WITH BECKMAN COULTER-TRAINED PERSONNEL BEFORE ATTEMPTING
TO OPERATE INSTRUMENT. DO NOT ATTEMPT TO PERFORM ANY PROCEDURE BEFORE CAREFULLY READING ALL
INSTRUCTIONS. ALWAYS FOLLOW PRODUCT LABELING AND MANUFACTURER’S RECOMMENDATIONS. IF IN DOUBT AS
TO HOW TO PROCEED IN ANY SITUATION, CONTACT YOUR BECKMAN COULTER REPRESENTATIVE.

HAZARDS AND OPERATIONAL PRECAUTIONS AND LIMITATIONS

WARNINGS, CAUTIONS, and IMPORTANTS alert you as follows:

WARNING - Can cause injury.

CAUTION - Can cause damage to the instrument.

IMPORTANT - Can cause misleading results.

BECKMAN COULTER, INC. URGES ITS CUSTOMERS TO COMPLY WITH ALL NATIONAL HEALTH AND SAFETY
STANDARDS SUCH AS THE USE OF BARRIER PROTECTION. THIS MAY INCLUDE, BUT IT IS NOT LIMITED TO,
PROTECTIVE EYEWEAR, GLOVES, AND SUITABLE LABORATORY ATTIRE WHEN OPERATING OR MAINTAINING THIS OR
ANY OTHER AUTOMATED LABORATORY ANALYZER.

WARNING Risk of operator injury if:

r All doors, covers and panels are not closed and secured in place prior to and during instrument operation.
r The integrity of safety interlocks and sensors is compromised.
r Instrument alarms and error messages are not acknowledged and acted upon.
r You contact moving parts.
r You mishandle broken parts.
r Doors, covers and panels are not opened, closed, removed and/or replaced with care.
r Improper tools are used for troubleshooting.

To avoid injury:

r Keep doors, covers and panels closed and secured in place while the instrument is in use.
r Take full advantage of the safety features of the instrument. Do not defeat safety interlocks and sensors.
r Acknowledge and act upon instrument alarms and error messages.
r Keep away from moving parts.
r Report any broken parts to your Beckman Coulter Representative.
r Open/remove and close/replace doors, covers and panels with care.
r Use the proper tools when troubleshooting.

CAUTION System integrity might be compromised and operational failures might occur if:

r This equipment is used in a manner other than specified. Operate the instrument as instructed in the Product Manuals.
r You introduce software that is not authorized by Beckman Coulter into your computer. Only operate your system’s

computer with software authorized by Beckman Coulter.

r You install software that is not an original copyrighted version. Only use software that is an original copyrighted

version to prevent virus contamination.

IMPORTANT If you purchased this product from anyone other than Beckman Coulter or an authorized Beckman Coulter

distributor, and, if it is not presently under a Beckman Coulter service maintenance agreement, Beckman Coulter cannot
guarantee that the product is fitted with the most current mandatory engineering revisions or that you will receive the most
current information bulletins concerning the product. If you purchased this product from a third party and would like
further information concerning this topic, call your Beckman Coulter Representative.

REVISION STATUS

Initial Issue A, 10/05

Software Version 1.0

Issue AA, 11/08

Software Version 1.0
Revision AA contains updates or new information in the following chapters:

r Chapter 1:

t Updates to the laser warning labels instructions, and graphic depicting location of all of

these labels. See Heading 1.6, WARNING LABELS AND PRECAUTIONS section.

t Updates to Table 1.1, List of Approved Reagents, under Heading 1.8, REAGENTS.

t Indicates the BCI website to access reagent information and search by Reagent Name or

Part Number, under Heading 1.8, REAGENTS.

t Indicates the BCI website to access Cell Lab Quanta System Application Notes, under

Heading 1.8, REAGENTS.

t Added instructions for three new warning labels:

t Heading 1.6, WARNING LABELS AND PRECAUTIONS

r Chapter 4:

t Updates to Table 4.7, Cups Approved for Use , under Heading 4.2, CUPS

Issue AB, 07/09

Software Version 1.0
Revision AB contains an update to the new corporate address.

Issue AC, 03/10

Software Version 1.0
Revision AC contains updates or new information in the following chapters:

r Cover page:

t Replaced the manufacturer’s symbol to show the Brea, California address.

r Chapter 1:

t Replaced the label on the rear of the instrument to show the Brea, CA address.

r Chapter 4:

t Updated Table 4.1, Absolute Count Performance Specifications under Heading 4.1,

PERFORMANCE SPECIFICATIONS.

r Chapter 7:

t Added an important notice under the Playing Back Files section.

t Updated the following sections: Create/Delete Protocol Groups, Assign a User to a

Protocol Group and Remove a User Assigned to a Protocol Group.

Issue AD, 10/11

Software Version 1.0.

Changes were made to pages 2-3.

Note: Changes that are part of the most recent revision are indicated in text by a bar in the
margin of the amended page.

This document applies to the latest software listed and higher versions. When a subsequent software version
affects the information in this document, a new issue will be released to the Beckman Coulter website. For
labeling updates, go to www.beckmancoulter.com and download the latest version of the manual or system
help for your instrument

PN 721742AD

iii

REVISION STATUS

iv

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REVISION STATUS, iii

CONTENTS, v

Illustrations, xi

Tables, xiii

INTRODUCTION, xv

Text Conventions, xvi
Graphic Conventions, xvi
Safety Symbols, xvii

1 USE AND FUNCTION,1-1

1.1 SYSTEM OVERVIEW,1-1

1.2 INTENDED USE,1-1

1.3 QUANTA SC SYSTEM OVERVIEW,1-2

CONTENTS

1.4 SYSTEM COMPONENTS,1-3

1.5 PRINCIPLES OF OPERATION,1-4

1.6 WARNING LABELS AND PRECAUTIONS,1-5
Mercury Arc Lamp,1-5
Laser,1-6

Laser Interlocks,1-8

Handling Precautions,1-9

1.7 OPTIONS,1-9
Hardware Options,1-9

Flow Cell Configurations,1-10
Jumper Positions for Different Size Particles µm,1-11
Light Source Configuration,1-11
Printer,1-11

1.8 REAGENTS,1-11

1.9 CONTROLS AND INDICATORS,1-12
Waste Bottle and Vacuum Bottle,1-13
PC,1-14

1.10 MATERIAL SAFETY DATA SHEETS (MSDS),1-15

PN 721742AD

2INSTALLATION,2-1

2.1 DELIVERY INSPECTION,2-1

2.2 SPECIAL REQUIREMENTS,2-1
Space and Accessibility,2-1
Installation Category,2-2

v

CONTENTS

Electrical Input,2-3
Ambient Temperature and Humidity,2-3
Heat Dissipation,2-3
Drainage,2-4

2.3 INSTALLATION PROCEDURES,2-4
Interunit Connections,2-4
Lifting and Carrying,2-4

3 OPERATION PRINCIPLES,3-1

3.1 OVERVIEW,3-1

3.2 SAMPLE FLOW,3-1

4 SPECIFICATIONS,4-1

4.1 PERFORMANCE SPECIFICATIONS,4-1
Absolute Count,4-1
Fluorescence,4-1
Sizing,4-1
Optics,4-1
Parameters,4-2
Calculated Parameters,4-2

FC and FSD Relevance,4-2
Mathematical Definitions,4-2
Numeric Range of FSD and FC,4-2
FSD and FC Calibration,4-3

Possible FSD and FC Calculation Errors,4-3
Software,4-3
Fluidics,4-3
Installation Requirements,4-4

4.2 CUPS,4-4

4.3 FLOW CELL,4-4

5 GETTING STARTED,5-1

5.1 POWERING UP THE SYSTEM,5-1
Turn On the Instrument and the Computer,5-1

5.2 INTRODUCTION TO THE QUANTA SC SOFTWARE,5-1
Overview,5-1
Microsoft Windows Basics,5-2

Desktop,5-2
Windows Password,5-2

Launching the Software,5-3

To Launch from the Start Menu,5-3
To Launch from the Desktop,5-3

Main Screen,5-4

Main Menu,5-4

vi

PN 721742AD

Menu Options,5-4

File Menu,5-5
Instrument Menu,5-5
Gain Menu,5-6
Vol um e M en u,5-6
Analysis Menu,5-6
Regions Screen,5-7
Compensation Screen,5-7
Protocols Screen,5-8
Current Settings Screen,5-8
File Information Screen,5-9
Security Menu,5-9
Help Menu,5-10

Closing a Window or Closing the Software,5-10

Closing a Window,5-10
Closing the Software,5-10

5.3 POWERING DOWN THE SYSTEM,5-11
Turn Instrument and Workstation OFF,5-11

CONTENTS

5.4 PLACING A CUP ON THE INSTRUMENT,5-12

6DAILY ROUTINE,6-1

6.1 PERFORM STARTUP,6-1

6.2 RUN QC (QUALITY CONTROL) SAMPLES,6-4

6.3 PERFORM SHUTDOWN,6-4

7SOFTWARE,7-1

7.1 UNDERSTANDING THE MAIN SCREEN,7-1
Parameter Information,7-3

Calibrate FSD or FC,7-4

Region Statistics,7-5

7.2 WORKING WITH THE FILE MENU,7-5
Saving Listmode (.LMD) Files,7-6
Auto Save Option,7-6
Playing Back Files,7-7
Defining Playback Options,7-8
Generating a Microsoft Excel Report,7-9
Exporting Data to Excel,7-9
Printing,7-9
Exiting (Closing) the Application,7-11

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7.3 WORKING WITH THE INSTRUMENT MENU,7-11
Start Up,7-12
Shut Down,7-13
Defining the Power Setting,7-14
Running the Cleaning Cycle,7-16

vii

CONTENTS

Flushing,7-17
Fill Cup,7-18
Resetting Fluid Count,7-18
Setting Up the Instrument,7-19

Understanding the Instrument Setup Screen,7-20
Understanding the Diagnostic Functions Screen,7-22

Laser Control,7-23

Understanding the Laser Control Options,7-23

Importing Settings,7-24

Understanding the Import Settings Screen,7-25

Automatically Aligning the Optics,7-25

Understanding the Auto Optical Alignment Screen,7-26

7.4 WORKING WITH THE GAIN MENU,7-27
Show Gain Settings,7-27
Defining Tracking Settings,7-28

Understanding the Tracking Settings Screen,7-29
Tracking Start,7-30
Reset Tracking,7-31

7.5 WORKING WITH THE VOLUME MENU,7-31
Calibrating the Volume,7-31

Calibration Beads,7-32
Understanding the Volume Calibration Screen,7-35

Displaying Channels,7-35

7.6 WORKING WITH THE ANALYSIS MENU,7-36
Parameter Ratio Analysis,7-36
Data Flag Settings,7-37

7.7 WORKING WITH THE REGIONS MENU,7-39
Managing Regions,7-39

Understanding the Manage Regions Screen,7-42

Showing Region Statistics,7-45

Understanding the Region Statistics Screen,7-46

7.8 WORKING WITH THE COMPENSATION MENU,7-47
Defining Compensation Settings,7-47

Understanding the Compensation Settings Screen,7-50

7.9 WORKING WITH THE MANAGE PROTOCOLS SCREEN,7-52

Understanding the Protocol Management Screen,7-53
Loading Protocols,7-54
Creating New Protocols,7-55
Saving Current Settings to a Protocol,7-57
Deleting Protocols,7-58
Save Description Changes (Editing a Protocol),7-60
Create/Delete Protocol Groups,7-61

Assign a User to a Protocol Group,7-62

Remove a User Assigned to a Protocol Group,7-63

viii

PN 721742AD

7.10 WORKING WITH THE CURRENT SETTINGS SCREEN,7-64
Defining Stop Sample Criteria,7-64
Defining the Concentration (Use, # of Seconds, and Start Time),7-65
Enabling/Disabling Checkboxes,7-65
Defining Auto Save Options,7-66
Customizing an Excel Report,7-68

Pre-defined Excel Templates,7-68
Creating/Modifying Text in Excel Report,7-68

Defining Filter Configurations,7-70

Understanding the Current Instrument Settings Screen,7-71

7.11 WORKING WITH THE FILE INFORMATION SCREEN,7-73
Entering Sample Information,7-74

Understanding the File Information Screen,7-75

7.12 WORKING WITH THE SECURITY MENU,7-76
Change Password,7-76
Manage Users,7-77

Understanding the Security Menu Screen,7-78
Password Options,7-79
Modify Database Path,7-79

CONTENTS

7.13 WORKING WITH THE HELP MENU,7-80
Launching Help,7-80
Viewing Software Information,7-80

8 QUALITY CONTROL,8-1

8.1 OVERVIEW,8-1

8.2 QC MATERIALS,8-1

8.3 DAILY QC,8-1
Before Running Flow-Check Fluorospheres,8-1
Running Flow-Check Fluorospheres,8-2
Running the Arc Lamp Alignment Beads,8-3

8.4 QC METHOD,8-4
Prepare QC Material,8-4

Flow-Check Fluorospheres,8-4

8.5 AUTOMATICALLY ALIGNING THE OPTICS,8-4

Arc Lamp Alignment Beads,8-4

9 SAMPLE ANALYSIS,9-1

9.1 BEFORE RUNNING SAMPLES,9-1

PN 721742AD

9.2 PREPARING SAMPLES,9-1

9.3 RUNNING SAMPLES,9-2

9.4 AFTER RUNNING SAMPLES,9-4

ix

CONTENTS

9.5 SETTING INITIAL GAIN, VOLTAGE AND DISCRIMINATOR VALUES,9-5

9.6 WORKING WITH REGIONS,9-5

10 CLEANING/REPLACEMENT,10-1

10.1 INSTRUMENT CLEANING AND HANDLING REQUIREMENTS, 10-1

10.2 INSTRUMENT QUICK DISCONNECT, 10-1

10.3 OPENING/CLOSING THE COVER, 10-2

10.4 EMPTYING THE WASTE BOTTLE, 10-4

10.5 CHANGING A FILTER, 10-5

10.6 CHANGING AN ARC LAMP EXCITATION FILTER, 10-12

10.7 REPLACING THE FUSE, 10-15

10.8 ADJUSTING FUSE VOLTAGE, 10-17

11 TROUBLESHOOTING,11-1

11.1 SYSTEM CONNECTIONS, 11-1

11.2 MONITOR VACUUM READINGS, 11-3

11.3 ERROR MESSAGES, 11-4

11.4 TROUBLESHOOTING GUIDE, 11-5

REFERENCES, REFERENCES-1

GLOSSARY,GLOSSARY-1

INDEX,INDEX-1

BECKMAN COULTER, INC. CUSTOMER END USER LICENSE AGREEMENT,1

x

PN 721742AD

Illustrations

1.1 Quanta SC System,1-1

1.2 Schematic Overview of Quanta Instrument,1-2

1.3 System Components,1-3

1.4 Triangular Flow Cell: Overview,1-4

1.5 Mercury Arc Lamp Warning Labels,1-5

1.6 Laser Warning Labels,1-7

1.7 Laser Warning Labels,1-7

1.8 Laser Interlocks,1-8

1.9 Optical Filter Configuration -- Laser 488 nm,1-9

1.10 Optical Filter Configuration -- Lamp 365 nm,1-10

1.11 Inside the Instrument: Controls and Indicators,1-12

1.12 Vacuum Regulator: Controls and Indicators,1-12

1.13 Jumper Positions: Controls and Indicators,1-13

1.14 Reservoirs: Controls and Indicators,1-13

1.15 Waste System and Reagents,1-14

1.16 Workstation PC: Controls and Indicators,1-14

5.1 Desktop,5-2

5.2 Main Screen,5-4

5.3 Main Menu,5-4

5.4 File Menu,5-5

5.5 Instrument Menu,5-5

5.6 Gain Menu,5-6

5.7 Volume Menu,5-6

5.8 Analysis Menu,5-6

5.9 Regions Screen,5-7

5.10 Compensation Screen,5-7

5.11 Protocols Screen,5-8

5.12 Current Instrument Settings Window,5-8

5.13 File Information Screen,5-9

5.14 Security Menu,5-9

5.15 Help Menu,5-10

7.1 Main Screen: Defined,7-1

7.2 Main Screen: Parameter Information,7-3

7.3 Main Screen: Calibrate,7-4

7.4 Main Screen: Region Statistics,7-5

7.5 Instrument Setup Screen: Defined,7-20

7.6 Diagnostic Functions Screen: Defined,7-22

7.7 Laser Control Options: Defined,7-23

7.8 Import Setting Screen: Defined,7-25

7.9 Auto Optical Alignment Screen: Defined,7-26

7.10 Tracking Settings Screen: Defined,7-29

7.11 Volume Calibration Screen: Defined,7-35

7.12 Manage Regions Screen: Defined,7-42

7.13 Single Parameter Region Definition Screen: Defined,7-43

7.14 Region Definition Screen for And/Or Regions: Defined,7-44

7.15 Region Statistics Screen: Defined,7-46

7.16 Compensation Settings Screen: Defined,7-50

7.17 Set Compensation using the FLn Control Screen: Defined,7-51

CONTENTS

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xi

CONTENTS

7.18 Protocol Management Screen: Defined,7-53

7.19 Current Instrument Settings Screen: Defined,7-71

7.20 File Information Screen: Defined,7-75

7.21 Security Menu: Manage Users,7-78

10.1 Disconnect Analyzer Power, 10-1

11.1 Cable Connections: Back of Instrument, 11-1

11.2 Cable Connections: Back of PC, 11-2

11.3 Cable Connections: Monitor, 11-2

11.4 Tubing Connections: Waste Bottle and Vacuum Bottle, 11-3

xii

PN 721742AD

Tables

1.1 List of Approved Reagents,1-11

2.1 System Dimensions and Accessibility,2-2

4.1 Absolute Count Performance Specifications,4-1

4.2 Fluorescence Performance Specifications,4-1

4.3 Sizing Performance,4-1

4.4 Optics Performance,4-1

4.5 Fluidics Specifications,4-3

4.6 Installation Requirements,4-4

4.7 Cups Approved for Use,4-4

11.1 Error Messages, 11-4

11.2 Troubleshooting Guide, 11-5

CONTENTS

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xiii

CONTENTS

xiv

PN 721742AD

OVERVIEW

This introductory section contains the following topics:

r ABOUT THIS MANUAL

r CONVENTIONS, and

r GRAPHICS.

ABOUT THIS MANUAL

The manual covers the description, installation, operation and maintenance of the Cell Lab

TM

Quanta

The information in your Quanta SC Instructions For Use manual is organized as follows:

s Chapter 1, USE AND FUNCTION

s Chapter 2, INSTALLATION

SC system.

Contains a short description of the major instrument components and options, and the
reagents and quality control materials used.

Contains instrument requirements, and diagrams of the interunit cable connections.

INTRODUCTION

s Chapter 3, OPERATION PRINCIPLES

Contains a brief description of how the system uses light scatter analysis to perform
cellular enumeration.

s Chapter 4, SPECIFICATIONS

Details the instrument and performance specifications.

r Chapter 5, GETTING STARTED

Provides information needed to get started, including Powering Up and Powering Down.

r Chapter 6, DAILY ROUTINE

Provides instructions for procedures that need to be done daily, including Start Up and
Shut Down.

r Chapter 7, SOFTWARE

Provides details on how to use the software.

r Chapter 8, QUALITY CONTROL

Provides information on how to run quality control material to verify instrument setup.

r Chapter 9, SAMPLE ANALYSIS

Provides information on how to run patient samples.

r Chapter 10, CLEANING/REPLACEMENT

Provides information on cleaning and replacement procedures.

r Chapter 11, TROUBLESHOOTING

Provides information on error messaging and instrument troubleshooting guide.

PN 721742AD

r INDEX

Provides page numbers for indexed information.

xv

INTRODUCTION

CONVENTIONS

CONVENTIONS

Text Conventions

r Bold font indicates a software option, such as Startup.

r Italics font indicates screen text displayed on the instrument, such as

r A Note contains supplemental information.

r An ATTENTION contains information that is important to remember or helpful when

r The terms “screen” and “window’ are used interchangeably.

r Quanta is used interchangeably with Quanta SC System and instrument.

r

Graphic Conventions

r indicates “select” with or “click” the left mouse button.

Run Shutdown.

performing a procedure.

Bold, italics font indicates a procedure heading.

r indicates double-click with the left mouse button.

r indicates “click” the right mouse button.

xvi

PN 721742AD

SYMBOLS

Safety Symbols

INTRODUCTION

SYMBOLS

Safety symbols alert you to potentially dangerous conditions. These symbols, together with
text, apply to specific procedures and appear as needed throughout this manual.

Symbol Warning Condition

Biohazard. Consider all materials (specimens,

reagents, controls, and calibrators, and so forth)
and areas these materials come into contact with as
being potentially infectious.

Electrical shock hazard.
shock when instrument is plugged in to the power
source.

Hot Surface hazard. Possibility of injury from a hot
surface.

Light hazard. Consider all light sources and light
emissions as being potentially hazardous to your
eyes.

Laser hazard. Consider all laser sources as being
potentially hazardous to your eyes.

Possibility of electrical

Action

Wear standard laboratory attire
and follow safe laboratory
procedures when handling any
material in the laboratory.

Before continuing, unplug the
instrument from the electrical
outlet.

Before continuing, use caution
when touching a surface that may
be hot.

Before continuing, verify that you
are wearing the proper protective
eye wear to avoid damage to your
eyes from beams of light. Never
look directly into a beam of light.

Before continuing, verify that you
are wearing the proper protective
eye wear to avoid damage to your
eyes from beams of light. Never
look directly into a beam of light.

GRAPHICS

PN 721742AD

Before continuing, verify that you
have read and understood the
warning described in the product
labeling.

Before continuing, please contact
your dealer or local Beckman
Coulter office for proper
decontamination information and
take back program to facilitate the
proper collection, treatment,
recovery, recycling, and safe
disposal of device.

A28219-AA

International warning. Whenever this symbol is
present, refer to the product labeling for detailed
description of the warning.

WEEE International warning. Whenever this symbol
is present, refer to local disposal requirements in
the event that the labeled parts need replacement or
disposal.

All graphics, including screens and printouts, are for illustration purposes only and must not
be used for any other purpose.

xvii

INTRODUCTION

GRAPHICS

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PN 721742AD

1.1 SYSTEM OVERVIEW

Figure 1.1 shows the Cell Lab Quanta SC system. The Quanta SC system is a three-color flow

cytometer, which provides the additional sizing parameter of Electronic Volume (EV) and the
granularity differentiating parameter of Side Scatter. Two light sources are provided; the
mercury arc lamp and the 488 nm solid state laser.

Figure 1.1 Quanta SC System

USE AND FUNCTION

1

1

1.2 INTENDED USE

The Cell Lab Quanta SC instrument is intended for General Laboratory Use. The instrument
simultaneously measures the fluorescence and electronic volume (EV) based on the Coulter

1

Principle.

PN 721742AD

1-1

USE AND FUNCTION

QUANTA SC SYSTEM OVERVIEW

1.3 QUANTA SC SYSTEM OVERVIEW

Figure 1.2 shows the major components of the Quanta instrument.

Figure 1.2 Schematic Overview of Quanta Instrument

Vacuum System

Vacuum

Pump

Vacuum

Bottle

COM2 (PC) Laser Control

Waste
Bottle

Controller

Power

Supply

Vacuum

Regulator

10 " Hg

Drip

Chamber

Fiber

Optics

Laser 488 nm

M

Valve

Assembly

Microscope

UG1 Filter

X Axis

Beam Shaper

M

Focus (F)

Dichroic

450 dcxr

M

Transducer’s

Flow Cell

Electrodes

Flow Cell

channel

Micro-objective
(100x) 1.25µm

Mirror

X Axis

position (T)

Y Axis

Diode

z 488 rdc

SC

525 BP (Laser)
460 BP (Lamp)

Sheath fluid

Sample cup

Filter Plate

550 DLP

Pre-Amp ECV

bottle

600 DLP

670 LP

FL2FL1

Syringe

Pump COM 1

(PC)

Power
Supply

Syringe Pump

PMT

FL3

575 BP

PMTPMT

Mercury Arc

Lamp 100 W

2380057

Power

Supply

M

Y Axis

Z Axis
(Focus)

Motors Motors (PC)

(Lamp’s motors & Microsc’s motors)

Log Amplifiers

Card

A/D

A/D
FL2

Motherboard

A/D
FL1

A/D

SC

EV

SIU Board (PC) Data

Syringe Pump

Cage

A/D
FL3

1-2

PN 721742AD

1.4 SYSTEM COMPONENTS

See Figure 1.3

Figure 1.3 System

b

c

d

USE AND FUNCTION

SYSTEM COMPONENTS

e

f

1

g

h

Components

Workstation Computer.

b

To control the cytometer and analyze data from
the cytometer.

Flow Cytometer.

d

Contains fluidics, optical flow chamber,
electronics, laser and arc lamp.

Vacuum Module.

f

Provides system vacuum.

Shutdown Solution.

h

Contains shutdown solution.

Workstation Monitor.

c

Displays data from the Workstation computer.

Waste Collection.

e

Contains Waste bottle and Vacuum bottle for
waste collection.

Sheath Container.

g

Contains sheath fluid.

PN 721742AD

1-3

USE AND FUNCTION

PRINCIPLES OF OPERATION

1.5 PRINCIPLES OF OPERATION

Electronic volume (EV) and optical measurements are made simultaneously in the same
spatial location consisting of a flow cell with an equilateral triangular cross section. See

Figure 1.4.

Figure 1.4 Triangular Flow Cell: Overview

This triangular flow geometry produces large hydrodynamic forces that focus the sample
stream to the center of the triangular aperture and allows the simultaneous collection of
optical and electronic volume measurements.

The optical system consists of a 100X oil immersion micro-objective with a numerical
aperture of 1.25 used to collect the fluorescence emission. A 100 W stabilized arc lamp with
wavelengths of 365, 404, and 435; and a 488 nm laser are used as the light sources for the
florescence measurements. The fluorescence signals are collected with photomultiplier tubes.

The fluidics system is controlled with a metering pump, which also consists of 17
computer-controlled valves that automatically perform all the sample handling and flushing
operations. Fluid transfer is accomplished by vacuum.

1-4

PN 721742AD

1.6 WARNING LABELS AND PRECAUTIONS

The Cell Lab Quanta SC system contains a 100-Watt Arc Lamp (Figure 1.5) emitting high
intensity heat and a 488 nm diode laser (Figure 1.6). The instrument, therefore, may pose
certain hazards associated with this lamp and laser if misused.

Mercury Arc Lamp

WARNING The Arc Lamp has power to 100 W and is accessible when the instrument cover is opened.

Avoid contact with the lamp.

When operating the instrument with the cover opened, coming into contact with the Arc Lamp may cause
severe burns. Avoid coming into contact with the lamp.

To avoid injury: Limit access to the inside of the instrument while the lamp is turned on to only those
occasions when there is an absolute need to open the instrument cover (Start Up or Calibration). Never look
directly into the lamp.

USE AND FUNCTION

WARNING LABELS AND PRECAUTIONS

1

Figure 1.5 Mercury Arc Lamp Warning Labels

10

0

20

10

20

90

40

40

80

90

40

40

70

50

80

50

60

60

70

PN 721742AD

1-5

USE AND FUNCTION

WARNING LABELS AND PRECAUTIONS

Laser

The Cell Lab Quanta SC is a Class I laser product.

WARNING Avoid direct exposure to the beam.

Reflected laser light can be as damaging as the original beam. Remove rings, watchbands, metal pens,
pendants and any other reflective accessories from hands and clothing.

Eye and skin exposure to direct and reflected laser light is hazardous and may be extremely harmful.

Ensure that all mirrors and optics are securely positioned and fixed. Prevent stray reflections from other
surfaces.

Do not place reflective objects in the laser beam.

Limit access to the laser to personnel who are familiar with the equipment. The laser must not be installed,
operated or repaired by inexperience or untrained personnel.

Do not open the Controller or Laser Head enclosure for any reason. Always return the units to the
manufacturer for repair.

Provide bright light around the laser equipment to reduce the operator’s pupil size.

Always wear eye protection appropriate to the beam wavelength and intensity when in the vicinity of the
laser equipment. Note: Glasses may make the beam invisible, increasing the risk of skin burns.

The laser equipment must be turned off when not in use.

Never operate the unit in the presence of flammable gases or fumes.

Laser radiation may be emitted from the end of the fiber optic cable. Never look directly into the fiber while
the laser is ON.

1-6

PN 721742AD

Figure 1.6 Laser Warning Labels

SCARED YA!

BOO!

L

L
M

M
S

S

USE AND FUNCTION

WARNING LABELS AND PRECAUTIONS

1

Figure 1.7 Laser Warning Labels

00000000

S/N

CELL LAB QUANTA SC

NAME

00000000

P/N

100/120/240

VOLTS

4 / 4 / 2

AMPS

50/60

HERTZ

SINGLE

PHASE

Manufactured for

Beckman Coutler, inc. by

CMSI

1313 Shotgun Road

Sunrise, Florida

U.S.A. 33326

PN 721742AD

1-7

USE AND FUNCTION

WARNING LABELS AND PRECAUTIONS

Laser Interlocks

WARNING Risk of personal injury if the laser safety interlock is bypassed. Do not tamper with the laser

interlock unless otherwise instructed in this manual.

Figure 1.8 shows the laser interlocks.

Figure 1.8 Laser Interlocks

Top cover

interlock

10

20

40

0

10

20

40

90

40

90

40

80

50

60

70

80

50

60

70

Front door

interlock

1-8

PN 721742AD

Handling Precautions

Proper handling procedures for samples and reagents used in flow cytometry analysis should
be adhered to at all times. Consult appropriate Material Safety Data Sheets for all diluents and
reagents used.

1.7 OPTIONS

Hardware Options

Figure 1.9 Optical Filter Configuration -- Laser 488 nm

USE AND FUNCTION

OPTIONS

1

PN 721742AD

1-9

USE AND FUNCTION

OPTIONS

Figure 1.10 Optical Filter Configuration -- Lamp 365 nm

1-10

Flow Cell Configurations

Description Configuration

Standard 125 x 125 µm

PN 721742AD

USE AND FUNCTION

REAGENTS

Jumper Positions for Different Size Particles µm

Recommended EV

Flow Cell

Aperture µm

125 3-40 15-40 10-15 3-10 Cells, large beads

Particle Size

limits µm

Jumper Position for different

Size Particles µm*

LMS

Suggested

particles to be analyzed

* Sample mean size is depicted in the above table for Electronic Volume (EV)

Light Source Configuration

Quanta SC Systems configuration contains both light sources, Mercury Arc Lamp and the
488 nm Laser with side scatter and three fluorescence parameters.

IMPORTANT Erroneous results can occur if both light systems (Mercury Arc Lamp and 488 nm Laser) are

active at the same time. Ensure that the appropriate light source is selected during the startup procedure,
refer to Heading 6.1, PERFORM STARTUP for detailed instructions.

1

Printer

A printer is not supplied with the instrument but is available as an option.

1.8 REAGENTS

Do not use any reagents that are not compatible with the specific wetted surfaces of the
sample instrument, such as non-aqueous solvents.

For a list of approved reagents for use on this system, see Table 1.1.

Table 1.1 List of Approved Reagents

Reagent Name Part Number

IsoFlow Sheath Fluid 1x10L 8546859

IsoFlow Sheath Fluid 4x1.8L 8547008

Marine Iso-Diluent 10 L 731088

Shutdown Solution 5 L 629968

Cleaning Solution Kit 629969

COULTER CLENZ 500 mL 8546929

COULTER CLENZ 5 L 8546930

COULTER CLENZ 10 L 8546931

PN 721742AD

1-11

USE AND FUNCTION

CONTROLS AND INDICATORS

1.9 CONTROLS AND INDICATORS

For details, see the following illustrations:

r Figure 1.11, Inside the Instrument: Controls and Indicators

r Figure 1.12, Vacuum Regulator: Controls and Indicators

r Figure 1.13, Jumper Positions: Controls and Indicators.

Figure 1.11 Inside the Instrument: Controls and Indicators

b

10

0

20

10

20

90

40

40

80

90

40

40

70

50

80

50

60

60

70

c

d

Mercury Arc Lamp voltage

b

display

Adjustment dial for Mercury

c

Arc Lamp power

Mercury Arc Lamp power

d

starter

Arc Lamp Excitation Filter

e

Figure 1.12 Vacuum Regulator: Controls and Indicators

b

g

e

f

1742015A

c

d

e

Vacuum regulator adjustment

b

Sheath

c

Air

d

Waste

e

Vacuum

f

Vacuum gauge

g

1-12

PN 721742AD

Figure 1.13 Jumper Positions: Controls and Indicators

10

0

0
2

0

10

2

90

40

40

80

0

90

4

40

70

50

8

50

60

0

60

7
0

S
C
A

R
E

B

D

O

Y

O

A

!

!

b

c

d

Waste Bottle and Vacuum Bottle

For details see the following illustrations:

USE AND FUNCTION

CONTROLS AND INDICATORS

Jumper Position S

b

Jumper Position M

c

Jumper Position L

d

1

r Figure 1.14, Reservoirs: Controls and Indicators

r Figure 1.15, Waste System and Reagents

Figure 1.14 Reservoirs: Controls and Indicators

b

c

d

e

\

Power ON/OFF for pump

b

Fuse

c

Power cord, connects to surge

d

protector

Connects to instrument vacuum

e

bottle

PN 721742AD

1-13

USE AND FUNCTION

CONTROLS AND INDICATORS

Figure 1.15 Waste System and Reagents

C

d

Waste Bottle

b

Vacuum Bottle

c

Sheath Fluid Bottle

d

b

PC

See Figure 1.16.

Figure 1.16 Workstation PC: Controls and Indicators

e

e

b

Shutdown Solution Bottle

e

Monitor

b

Keyboard

c

d

c

1.44 Floppy Drive

d

CDRW Drive

e

CD-ROM Read/Write Drive

1-14

PN 721742AD

1.10 MATERIAL SAFETY DATA SHEETS (MSDS)

To obtain an MSDS for reagents used on this system:

1. On the internet, go to http://www.beckmancoulter.com:

a. Select MSDS from the Customer Support drop-down menu.

b. Follow the instructions on the screen

c. Contact your Beckman Coulter Representative if you have difficulty locating the

information.

2. If you do not have internet access:

r In the USA, either call Beckman Coulter Customer Operations (800-526-7694) or

write to:

Beckman Coulter, Inc.
Attention: MSDS Requests
P.O. BOX 169015
Miami, FL 33116-9015

r Outside the USA, contact your Beckman Coulter Representative.

USE AND FUNCTION

MATERIAL SAFETY DATA SHEETS (MSDS)

1

PN 721742AD

1-15

USE AND FUNCTION

MATERIAL SAFETY DATA SHEETS (MSDS)

1-16

PN 721742AD

2.1 DELIVERY INSPECTION

The instrument is tested before shipping. International symbols and special handling
instructions are printed on the shipping cartons to inform the carrier of the precautions and
care applicable to electronic instruments.

CAUTION Possible instrument damage could occur if you uncrate the instrument, install it, or set it up.

Keep the instrument in its packaging until your Beckman Coulter Representative uncrates it for installation
and setup.

When you receive your instrument, carefully inspect all cartons. If you see signs of
mishandling or damage, file a claim with the carrier immediately. If separately insured, file the
claim with the insurance company.

2.2 SPECIAL REQUIREMENTS

Before your system is installed, determine where you want the system placed. Consider the
following:

r Space and Accessibility

r Installation Category

r Electrical Input

INSTALLATION

2

2

r Ambient Temperature and Humidity

r Heat Dissipation

r Drainage

Space and Accessibility

Allow room to interconnect the system components. Consider:

r A comfortable working height

r Adequate space for ventilation, and access for maintenance and service

r Placement of the system so that:

r the vents are not blocked and proper airflow is allowed

r the system is at least 6 inches from any wall or object, such as a filing cabinet or

other instrument.

r Proper electrical requirements in the location chosen for system placement. To prevent

electrical shock, be sure to plug equipment into properly grounded electrical outlets.

r Once the computer and the system have been setup, store the shipping boxes in a dry

area.

PN 721742AD

2-1

INSTALLATION

SPECIAL REQUIREMENTS

Table 2.1 System Dimensions and Accessibility

Specifications Instrument

Workstation
Monitor

Workstation
Computer*

Height

Clearance above for servicing
Total clearance needed

Width

Clearance on right for servicing
Clearance on left for servicing
Total clearance needed

Depth

Clearance behind instrument for
sufficient cooling and room for
servicing

Total clearance needed

Weight 34.9 kg (77 lbs) 5.6 kg (12.4 lbs) 5.0 kg (11 lbs)

Sound Pressure Level <85 dBA <85 dBA <85 dBA

*Workstation Computer must be placed on the floor.

44.5 cm (17.5 in.)

48.3 cm (19 in.) min.

92.3 cm (36.5 in.)

55.9 cm (22 in.)

15.2 cm (6 in.)

15.2 cm (6 in.) min.

86.3 cm (34 in.)

70.5 cm (27.8 in.)

15.2 cm (6 in.)

85.7 cm (33.8 in.)

43.3 cm (17.1 in.)
N/A

43.3 cm (17.1 in.)

42.2 cm (16.6 in.)
N/A
N/A

42.2 cm (16.6 in.)

17.6 cm (6.9 in.)

N/A

17.6 cm (6.9 in.)

43.18 cm (17 in.)

20.32 cm (8 in.)

43.18 cm (17 in.)

Note: Keep all shipping containers in case your system needs to be shipped to Beckman
Coulter, Inc. for repair.

Installation Category

Category II (per IEC 1010-1 standard). Pollution degree 2.

2-2

PN 721742AD

INSTALLATION

SPECIAL REQUIREMENTS

Electrical Input

CAUTION Possible instrument damage could occur if you plug the Quanta SC System on the same

electrical circuit as another instrument or use an extension cord or a power strip to connect the Quanta SC
System. Use a dedicated outlet with isolated ground for the Quanta SC System plug.

2

AC Input Specifications
Instrument

AC Input Specifications
Pump Station

AC Input Specifications
Computer

Voltage Rating: 100/120 VAC Voltage Rating: 220/240 VAC

Current Rating: 6 A Current Rating: 3 A

Frequency Rating: 50/60 Hz Frequency Rating: 50/60 Hz

Connection Type: IEC 320/C14 Connection Type: IEC 320/C14

Voltage Rating: 100/120 VAC Voltage Rating: 220/240 VAC

Current Rating: 6 A Current Rating: 3 A

Frequency Rating: 50/60 Hz Frequency Rating: 50/60 Hz

Connection Type: IEC 320/C14 Connection Type: IEC 320/C14

Voltage Rating: 100/120 VAC Voltage Rating: 220/240 VAC

Current Rating: 6 A Current Rating: 3 A

Frequency Rating: 50/60 Hz Frequency Rating: 50/60 Hz

Connection Type: IEC 320/C14 Connection Type: IEC 320/C14

PN 721742AD

Ambient Temperature and Humidity

This device is intended for indoor use only.

Keep the room temperature between 16°C and 29°C (60°F and 84°F), and do not let it change
more than 3°C (5°F) since the last alignment verification. Keep the humidity between 30%
and 80%, without condensation. Maximum Altitude 2000m.

Heat Dissipation

Heat dissipation is 500 W (1,706 Btu/hour) for the total system. Provide sufficient air
conditioning (refer to Ambient Temperature and Humidity).

2-3

INSTALLATION

INSTALLATION PROCEDURES

Drainage

WARNING Risk of biohazardous contamination if you have skin contact with the waste container or

vacuum bottle, its contents, and its associated tubing. The waste container and its associated tubing might
contain residual biological material and must be handled with care. Clean up spills immediately. Dispose of
the contents of the waste container in accordance with your local regulations and acceptable laboratory
procedures.

The waste line from the Cytometer is connected to a waste bottle, which sits next to the
vacuum module. Dispose of the waste in accordance with your local environmental
regulations and acceptable laboratory procedures.

2.3 INSTALLATION PROCEDURES

Your system will be installed by a Beckman Coulter Representative.

Interunit Connections

Laser Power

Supply

Monitor

Computer

WASTE

VACUUM

Pneumatics SolutionsInstrument

Lifting and Carrying

WARNING Possible operator injury. One person only should not lift the instrument. Lifting handles are not

provided and the instrument weighs over 40 lbs. Lifting of the instrument should only be done by a
minimum of two persons following the requisite safety precautions.

2-4

PN 721742AD

3.1 OVERVIEW

The Quanta SC System is a three-color flow cytometer, which provides the additional sizing
parameter of Electronic Volume (EV), and the granularity differentiating parameter of side
scatter.

The Quanta SC System uses a triangular flow cell (Figure 1.4) filled with conductive sheath
liquid. Cells suspended in a weak electrolyte solution are drawn through a small aperture that
separates two electrodes between which a slight current flows. As each cell passes through the
electrical sensing zone of the aperture, the cell displaces its own volume of conducting liquid,
momentarily increasing the impedance. This change in resistance produces a voltage pulse
large enough to accurately measure; the Coulter Principle states that the amplitude of this
pulse is directly proportional to the volume of the cell. The Coulter Principle, which is not
affected by shape, color, or refractive index, is the accepted reference method for cell and
particle counting and sizing.

A lamp or laser light source is used to excite fluorescent dyes attached to the particles. This
light is directed toward the aperture in the flow cell.

As particles pass through the light source, the dye emission is reflected to a mirror and then to
filters that direct the fluorescent light in turn to three PMTs (FL1, FL2 and FL3). The PMTs
generate voltage pulses proportional to the amount of fluorescence.

OPERATION PRINCIPLES

3

3

If the laser is ON, then laser light bounces off the particles as they pass through the laser light.
The light which bounces off at a right angle is passed to the side scatter detector. This
detector also generates a voltage pulse proportional to the amount of light which bounced off
the particle.

The Electronic Volume (EV) pulse, fluorescent pulse(s) and the side scatter pulse are
amplified, digitized and then analyzed for pulse height. The pulse height of each
measurement is then used to create the plots seen on the main screen.

A suspension of cells or particles, of precise volume, is drawn through the orifice of an
aperture. A slight current is maintained, by two electrodes, across the orifice. Single cell
suspensions enter the “sensing zone” resulting in an increase in resistance ultimately
producing a voltage pulse proportional to the volume of the cell or particle. This
measurement is not affected by color, shape, or refractive index of the sample. The Coulter
Principle is the reference method for automated cell counting and sizing.

3.2 SAMPLE FLOW

When a sample is placed on the Cell Lab Quanta SC instrument and the Start button is
selected, the metering pump aspirates the sample from the sample cup into the sample loop
(holding station for the sample). Once the sample is in the sample loop, the sample is then
boosted from the sample loop to the entrance of the flow cell.

When the sample is at the entrance of the flow cell, the metering pump slows the sample
down to allow the triangulating forces of the sheath flow to line the particles in the sample up
as they pass through the flow cell. Once a stable flow has been achieved, the software begins
data collection.

PN 721742AD

3-1

OPERATION PRINCIPLES

SAMPLE FLOW

Data collection continues until the user manually stops it, a user defined stop criteria is met,
or the volume of sample that was aspirated into the sample loop has been dispensed through
the flow cell.

After a stop criteria is met or the sample volume has been dispensed, several automated post
sample processes may occur. These can include:

r Saving the file to disk

r Printing a file report

r Recovering the remaining sample into sample cup

r Rinsing the fluidics to prepare for the next sample.

3-2

PN 721742AD

4.1 PERFORMANCE SPECIFICATIONS

Absolute Count

Table 4.1 Absolute Count Performance Specifications

Description Specification

Accuracy ± 5%

Reproducibility (CV) < 5%

Concentration

Fluorescence

Table 4.2 Fluorescence Performance Specifications

Description Specification

4

3 x 10

to 2 x 106 particles per mL.

ATTENTION: For greatest accuracy, samples should have a

concentration in the range of 2.5 x 10

SPECIFICATIONS

4

5

to 2 x 106 particles per mL.

4

≤ 2.5% HPCV using TRBC DNA Reference Calibrator in linear scale with

Resolution

NIM-DAPI and the mercury arc lamp

≤ 3% HPCV using COULTER Flow-Check Fluorospheres in linear scale
with the 488 laser

Sizing

Table 4.3 Sizing Performance

Description Specification

Sizing Electronic impedance with a measurement range of 3.0 to 40 microns

diameter with the standard 125 µm flow cell.

Optics

Table 4.4 Optics Performance

Description Specification (in nms)

Excitation* 365, 404 or 435 arc excitation and a 488 nm laser

*May only operate one light source at a time.

Fluorescence Detectors 3 Ultra Sensitive Photomultipliers tube

Optical Alignment Automated Computer Controlled

PN 721742AD

Optical Coupling 1.25 NA oil immersion objective

Scatter Solid State Photo Diode

4-1

SPECIFICATIONS

PERFORMANCE SPECIFICATIONS

Parameters

r Fluorescence 1 Linear/Log display

r Fluorescence 2 Linear/Log display

r Fluorescence 3 Linear/Log display

r Side Scatter Linear/Log display

r EV Linear display

r Time display

r Counts per mL display

r Calculated Fluorescence Surface Density (FSD)/ Fluorescence Concentration (FC)

display

Calculated Parameters

The following parameters take advantage of the Coulter Volume used in Flow Cytometry:

r FC – Fluorescent Concentration

r FSD – Fluorescent Surface Density

FC and FSD are parameters calculated for every data point, and the calculation is treated as a
parameter for display, as well as statistics.

FC and FSD Relevance

FC and FSD are relevant whenever the quantity of stain is proportional to the size of the
particle (that is, when a bigger particle will allow for more staining than a smaller particle). If
this is the case, then FC (used for internal markers) or FSD (used for external markers) can
normalize the fluorescence to stain density, instead of total quantity of stain.

When the quantity of stain is proportional to the size of the particle the stain density (FC and
FSD) can help to separate populations that blend together using the total fluorescent
measurement.

Mathematical Definitions

FC = Fluorescent Channel/Volume Channel
FSD = Fluorescent Channel/(Volume Channel)^(2/3)

To get Surface Area of a Sphere from the Volume:

Volume = 4/3Pi R^3
Surface Area = 4Pi R^2
Surface Area = Volume^(2/3) * 4Pi*(3/(4*Pi)) ^ (2/3)

4-2

The surface area is equal to 2/3 root of the volume times a constant. For arbitrary channel
assignment, constants can be ignored as they are equivalent to gain.

Numeric Range of FSD and FC

The range of FC and FSD parameters is the addition of the range of the Fluorescent and
Volume parameters which are the basis of the calculation.

PN 721742AD

SPECIFICATIONS

PERFORMANCE SPECIFICATIONS

FC range = 1/EV max channel to FL Max Channel
FSD range = 1/(EV max channel)^2/3 to FL Max Channel

If EV is 1000 linear channels, and FL is 4 decades of Log then:
FC range = .001 to 10000 (7 decades)
FSD range = .01 to 10000 (6 decades)

FSD and FC Calibration

The FSD parameter can be calibrated to Antigen Density assuming that the stain density is
directly proportional to the Antigen Density.

The FC parameter can be calibrated for Absolute Concentration (the concentration of the
item that the internal marker binds to). This also assumes that the stain concentration is
proportional to the concentration of the item being calibrated.

The FC parameter may also be calibrated for NPE (Nuclear Packing Efficiency). This is the
inverse of the Absolute Concentration, and is specific for Cell Nuclei.

Possible FSD and FC Calculation Errors

The FSD parameter assumes the particle is spherical when calculating the surface area. If the
particle is not spherical, then the ratio of the volume to the surface area may not match the
calculated parameter which uses the 2/3 root of the volume to calculate the surface area.

4

Software

r Microsoft® Windows® XP Professional

r Microsoft Office

r FCS 2.0 Generated Files

Fluidics

Table 4.5 Fluidics Specifications

Description Specification

Flow Cell Patented 125 µm triangular flow cell (standard)

Sample Rate 4.17 µL to 100 µL per minute for 125 µm

Valves Computer-controlled fluidics system

Delivery System Vacuum pump/motorized metering pump

Minimum Sample Size 150 µL

Maximum Sample Size 2 mL

Carryover < 1% from one specimen to another when # of gated events is between

100 and 10,000.

PN 721742AD

4-3

SPECIFICATIONS

CUPS

Installation Requirements

Table 4.6 Installation Requirements

Description Specification

Power 100 Vac, 120 Vac, or 220 Vac, 50/60 cycles

Wattage Consumed 500 watts (total system)

Operating Temperature between 16°C and 29°C (60°F and 84°F)

Physical Dimensions 22 W x 28 D x 18 H (in.)

Instrument Weight* 34.9 kg (77 lbs)

4.2 CUPS

The Quanta SC System analyzes samples in cups.

Beckman Coulter does not recommend the use of one cup in preference to another nor
guarantees the acceptability of the sample cup to produce quality results. If you need
information on a sample cup not listed here, contact your Beckman Coulter Representative.

56 W x 71 D x 45 H (cm)

* Workstation weight is separate and varies with the workstation components manufacturers.

Table 4.7 Cups Approved for Use

Cup Size Manufacturer Part Number

4 mL
Note: Maximum Sample Volume, 2 mL

4.3 FLOW CELL

The standard flow cell is 125 µm.

Beckman Coulter, Inc.

383721

4-4

PN 721742AD

5.1 POWERING UP THE SYSTEM

Turn On the Instrument and the Computer

1 Power ON the Workstation computer.

2 Launch the Cell Lab Quanta SC

software.

3 If the Sign-In window appears, select

the User ID from the drop-down list
and enter password. For additional
information regarding User IDs and
passwords, refer to Heading 7.12,

WORKING WITH THE SECURITY
MENU.

GETTING STARTED

5

5

4 Run the startup item from the Instrument Menu, and follow the steps as prompted. Refer

to Heading 6.1, PERFORM STARTUP for detailed instructions regarding instrument
startup.

5.2 INTRODUCTION TO THE QUANTA SC SOFTWARE

Overview

Cell Lab Quanta SC software is compatible with Microsoft Windows XP based systems.
The Analysis software functions include:

r Data Collection

r Instrument Control

r Data Analysis

r Acquisition

r Reporting

r Gating

PN 721742AD

5-1

GETTING STARTED

INTRODUCTION TO THE QUANTA SC SOFTWARE

r Statistical Analysis

r Alignment

r Listmode Playback

r Calculate EV

r Printing

r Compensation

Microsoft Windows Basics

Desktop

After your Workstation is powered up and runs through its routines, the Windows desktop
(Figure 5.1) appears. Because desktops can be customized, yours may look different.
Beckman Coulter, Inc. recommends that the Windows scheme set up on your Workstation at
installation be maintained and not altered.

Figure 5.1 Desktop

5-2

Windows Password

The Cell Lab Quanta SC software is password protected and log on specific which protects
against unauthorized changes to your data or system. This protection is provided at the
application level only and does not apply to the Microsoft Windows XP Operating System
(OS).

Beckman Coulter recommends that you utilize the Microsoft Windows password and screen
saver password if you are concerned about protecting unauthorized changes to data files as
well as OS settings. Refer to your Microsoft Windows documentation or contact your

PN 721742AD

GETTING STARTED

INTRODUCTION TO THE QUANTA SC SOFTWARE

Network Administrator should you have difficulties setting the appropriate permissions for a
user ID.

Launching the Software

You can either launch the software from the Start menu or from your Windows desktop.

To Launch from the Start Menu

1. .

5

2.
appears.

All Programs tt Cell Lab Quanta SC tt Cell Lab Quanta SC. The Main screen (Figure 5.2)

PN 721742AD

To Launch from the Desktop

From the Windows desktop, . The Main screen (Figure 5.2) appears.

5-3

GETTING STARTED

INTRODUCTION TO THE QUANTA SC SOFTWARE

Main Screen

When you launch the software, the Main screen (Figure 5.2) appears. The Main menu
(Figure 5.3) appears at the top of the screen. For details about the Main screen, see

Heading 7.1, UNDERSTANDING THE MAIN SCREEN.

Figure 5.2 Main Screen

Main Menu

See Figure 5.3.

Figure 5.3 Main Menu

Menu Options

The Main menu options include:

r File Menu

r Instrument Menu

r Gain Menu

r Vol um e M en u

r Analysis Menu

r Regions Screen

r Compensation Screen

r Protocols Screen

r Current Settings Screen

5-4

PN 721742AD

r File Information Screen

r Security Menu

r Help Menu

File Menu

GETTING STARTED

INTRODUCTION TO THE QUANTA SC SOFTWARE

5

When you
see Heading 7.2, WORKING WITH THE FILE MENU.

Figure 5.4 File Menu

Instrument Menu

When you
menu, see Heading 7.3, WORKING WITH THE INSTRUMENT MENU.

File from the Main menu (Figure 5.4), appears. For details about this menu,

Instrument from the Main menu, Figure 5.5 appears. For details about this

PN 721742AD

Figure 5.5 Instrument Menu

5-5

GETTING STARTED

INTRODUCTION TO THE QUANTA SC SOFTWARE

Gain Menu

When you

Gain from the Main menu, Figure 5.6 appears. For details about this menu, see

Heading 7.4, WORKING WITH THE GAIN MENU.

Figure 5.6 Gain Menu

Volume Menu

When you

Volume from the Main menu, Figure 5.7 appears. For details about this menu,

see Heading 7.5, WORKING WITH THE VOLUME MENU.

Figure 5.7 Volume Menu

Analysis Menu

When you

Analysis from the Main menu, Figure 5.8 appears. For details about this menu,

see Heading 7.6, WORKING WITH THE ANALYSIS MENU.

Figure 5.8 Analysis Menu

5-6

PN 721742AD

Regions Screen

GETTING STARTED

INTRODUCTION TO THE QUANTA SC SOFTWARE

5

When you
see Heading 7.7, WORKING WITH THE REGIONS MENU.

Figure 5.9 Regions Screen

Compensation Screen

Regions from the Main menu, Figure 5.9 appears. For details about this menu,

When you
menu, see Heading 7.9, WORKING WITH THE MANAGE PROTOCOLS SCREEN.

Figure 5.10 Compensation Screen

Compensation from the Main menu, Figure 5.11 appears. For details about this

PN 721742AD

5-7

GETTING STARTED

INTRODUCTION TO THE QUANTA SC SOFTWARE

Protocols Screen

When you

Protocols from the Main menu, Figure 5.11 appears. For details about this

menu, see Heading 7.9, WORKING WITH THE MANAGE PROTOCOLS SCREEN.

Figure 5.11 Protocols Screen

Current Settings Screen

When you

Current Instrument Settings from the Main menu, Figure 5.12 appears. For details

about this menu, see Heading 7.10, WORKING WITH THE CURRENT SETTINGS SCREEN.

Figure 5.12 Current Instrument Settings Window

5-8

PN 721742AD

File Information Screen

GETTING STARTED

INTRODUCTION TO THE QUANTA SC SOFTWARE

5

When you
this menu, see Heading 7.11, WORKING WITH THE FILE INFORMATION SCREEN.

Figure 5.13 File Information Screen

File Information from the Main menu, Figure 5.13 appears. For details about

Security Menu

When you
menu, see Heading 7.12, WORKING WITH THE SECURITY MENU.

Figure 5.14 Security Menu

Security from the Main menu, Figure 5.15 appears. For details about this

PN 721742AD

5-9

GETTING STARTED

INTRODUCTION TO THE QUANTA SC SOFTWARE

Help Menu

When you

Help from the Main menu, Figure 5.15 appears. For details about this menu,

see Heading 7.13, WORKING WITH THE HELP MENU.

Figure 5.15 Help Menu

Closing a Window or Closing the Software

Throughout this manual, you may be instructed to close a software window or to close
(shutdown) the Quanta software. For details, see:

r Closing a Window

r Closing the Software

Closing a Window

To close an open software window and leave the software open, on the window to
close. The previous window appears.

Closing the Software

To close the Quanta software entirely and return to the Windows desktop, from the
Main screen. Your Windows desktop appears.

5-10

PN 721742AD

5.3 POWERING DOWN THE SYSTEM

Turn Instrument and Workstation OFF

1 Prior to powering down the system, perform shutdown procedure, see Heading 6.3,

PERFORM SHUTDOWN.

2 Close the Quanta software (see Closing the Software.)

3 At the Workstation, tt Shutdown tt Shutdown or TURN OFF.

GETTING STARTED

POWERING DOWN THE SYSTEM

5

4 Allow the system time to shut down.

5 Turn off the PC and monitor.

PN 721742AD

5-11

GETTING STARTED

PLACING A CUP ON THE INSTRUMENT

5.4 PLACING A CUP ON THE INSTRUMENT

Perform this procedure to place a sample cup securely on the instrument at the sample stage
(Figure ).

Sample Stage

WARNING Risk of contact with biohazardous

material if you use a cup not approved for use
on this instrument. Only use approved cups
listed in Table 4.7.

5-12

PN 721742AD

1 Place the cup on the instrument:

a. Introduce cup so that the probe

(

b) is inserted in the cup.

b. Tilt the cup and push up so that

the cup (
holder.

c) is secure in the

b

GETTING STARTED

PLACING A CUP ON THE INSTRUMENT

c

5

c. Push up until the cup is

completely secure.

2 When you are certain the cup is secure, release it so that it is supported by the holder.

PN 721742AD

5-13

GETTING STARTED

PLACING A CUP ON THE INSTRUMENT

5-14

PN 721742AD

6.1 PERFORM STARTUP

Perform this procedure at the beginning of each day and after switching between light sources
or filters.

1 Verify correct filters are installed for the selected application and light source.

2 Power ON the Workstation computer.

3 Launch the Cell Lab Quanta SC

software.

DAILY ROUTINE

6

6

4 Once the Quanta software has

launched, the following screen appears
if a username and password are
required. For additional information
regarding User IDs and passwords,
refer to Heading 7.12, WORKING

WITH THE SECURITY MENU.

5 Instrument tt Start Up. The following

screen appears.

PN 721742AD

6-1

DAILY ROUTINE

PERFORM STARTUP

6 Turn ON the Quanta instrument.

7 Next Step. The following screen

displays. Follow the directions on the
screen to continue with the Start Up
Procedure to startup the instrument.

For additional information regarding
this Start Up step, refer to

Heading 10.4, EMPTYING THE
WASTE BOTTLE and Heading 11.2,
MONITOR VACUUM READINGS.

8 Next Step. The following screen

displays. Follow the directions on the
screen to continue with the Start Up
Procedure to startup the instrument.

Refer to Figure 1.15, Waste System and

Reagents to locate the appropriate

containers.

6-2

PN 721742AD

PERFORM STARTUP

9 Next Step. The following screen

displays. Follow the directions on the
screen to continue with the Start Up
Procedure to startup the
instrument.

CAUTION Possible damage to the instrument if Mercury Arc Lamp power starter is continuously

depressed after visible light is evident. Do not continue to depress the Mercury Arc Lamp power
starter after observing visible light.

DAILY ROUTINE

6

CAUTION Possible damage or reduced bulb life expectancy if Mercury Arc Lamp is ignited when lamp

is HOT. If the Mercury Arc Lamp is turned OFF for any reason, including brief power failures, the lamp
must be allowed to cool before reignition. Wait at least 15-20 minutes before attempting to reignite
Mercury Arc Lamp. Very high or unstable CVs or HPCVs may occur as the result of a damaged Mercury
Arc Lamp.

IMPORTANT Possible erroneous results if laser is operated with a laser base temperature >7oC (45oC)

o

above ambient. Operating the laser at a temperature >7
HPCVs and CVs and/or the gain and voltage may have to be adjusted significantly from original
settings.

IMPORTANT Erroneous results can occur if both light systems are active at the same time. Ensure

that the appropriate light source is selected during the startup procedure.

C (45oC) above ambient may result in high

PN 721742AD

6-3

DAILY ROUTINE

RUN QC (QUALITY CONTROL) SAMPLES

10 Next Step. The following screen

displays. Follow the directions on the
screen to continue with the Start Up
Procedure to startup the instrument.

6.2 RUN QC (QUALITY CONTROL) SAMPLES

Beckman Coulter recommends the use of QC materials to gauge instrument performance of
reportable parameters. See Chapter 8, QUALITY CONTROL for details.

6.3 PERFORM SHUTDOWN

Perform this procedure at the end of each day.

Note: COULTER CLENZ can be used as the shutdown fluid if the instrument will be
shutdown for a time period not to exceed one (1) day.

CAUTION Risk of damage to the instrument if you exit the Quanta application or turn off the instrument

and computer without performing the Shutdown procedure. Perform the Shutdown procedure before
you exit the Quanta application or turn off the instrument and computer to prevent damage to the
instrument.

.

6-4

PN 721742AD

1 Instrument tt ShutDown. The

following screen displays. Follow the
directions on the screen to continue
with the Shut Down Procedure to
shutdown the instrument.

Refer to Figure 1.14, Reservoirs:

Controls and Indicators to locate the

appropriate containers.

CAUTION Instrument damage can occur if

instrument is shut down for more than one
(1) day using COULTER Clenz as the
shutdown solution. Use Shutdown Solution,
referenced in Table 1.1, List of Approved

Reagents.

DAILY ROUTINE

PERFORM SHUTDOWN

6

2 Next Step. The following screen

displays. Follow the directions on the
screen to continue with the Shut Down
Procedure to shutdown the instrument.

Refer to Figure 1.14, Reservoirs:

Controls and Indicators to locate the

appropriate containers.

PN 721742AD

6-5

DAILY ROUTINE

PERFORM SHUTDOWN

3 Next Step. The following screen

displays. Follow the directions on the
screen to continue with the Shut Down
Procedure to shutdown the instrument.

Refer to Figure 1.14, Reservoirs:

Controls and Indicators to locate the

appropriate containers and

Heading 10.4, EMPTYING THE
WASTE BOTTLE for proper waste

removal.

4 Next Step. The following screen

displays. Follow the directions on the
screen to continue with the Shut Down
Procedure to shutdown the instrument.

6-6

PN 721742AD

5 Turn OFF the Quanta instrument.

DAILY ROUTINE

PERFORM SHUTDOWN

6

PN 721742AD

6-7

DAILY ROUTINE

PERFORM SHUTDOWN

6-8

PN 721742AD

7.1 UNDERSTANDING THE MAIN SCREEN

The Main screen (Figure 7.1) can display up to ten graphs which may be either single or dual
parameter. The seven graph display includes statistics under each graph for its corresponding
regions. The X-axis of the first display is always the primary (trigger) parameter. The
following parameters are available for collection:

r FL1 (Fluorescent Channel 1)

r FL2 (Fluorescent Channel 2)

r FL3 (Fluorescent Channel 3)

r SS (Side Scatter)

r EV (Electronic Volume)

r Time (must be selected on the Current Setting screen; see Heading 7.10, WORKING

WITH THE CURRENT SETTINGS SCREEN); only displayed in the dual-parameter

scatterplots.

Figure 7.1 Main Screen: Defined

SOFTWARE

7

7

PN 721742AD

7-1

SOFTWARE

UNDERSTANDING THE MAIN SCREEN

Main menu (see Figure 5.3)

b

Zoom button: Show pop-up screens that

d

enlarge the display of the histograms and the
scatterplots. Use the slide bar on the left side to
adjust the Y-axis scale for single-parameter
histograms.

Display areas for the parameter selected from

f

the parameter drop down list.
The Main screen can display up to 10 graphs,

each of which may be single or dual
parameters, see Setting Up the Instrument for
detailed instructions.

Region statistics, gain and tracking windows

h

are displayed in this area.

Parameter Information button displays the

j

Parameter Information dialog which allows
users to customize certain parameter
properties (Name, Color, etc.). For additional
information about this dialog, see Parameter

Information.

Select a region to be the gate for the display

c

or select All Data Points. If a region is
selected from the drop down list, then only
data points within that region are shown on
the display.

Message area that displays the Laser state,

e

Compensation state, Data Flags and Protocol
Name, as appropriate.

Note: If the settings are modified after a
protocol is loaded, (modified) appears after
the protocol name.

Region statistics for the display directly

g

located above area.

Flow Rate Control bar: Controls the flow rate

i

of the sample through the instrument. If the
box is checked beside the bar, the instrument
automatically adjusts the flow rate to keep
the count rate constant during the sample
run.

Clear Data button resets the total count and

1)

erases all data points collected up to that
point. This should only be done if the data
collected is no longer desired. The rinse
automatically does this between samples.

7-2

Save button saves the listmode data file in FCS

1!

2.0 format to the operating system.

Rinse button cleans the fluidics and prepares

1#

the instrument for the next sample.

Aspirates sample into the instrument and

1@

begins data collection.

Lower Level Discriminator (LLD)/Upper Level

1$

Discriminator (ULD) bar: activates the scroll
bar allowing you to adjust the selected
discriminator value. Setting these values is
important for every experiment.

Coordinate discriminator values with gain
adjustments on the front of the instrument.

In most cases, the LLD should be set to a
level where the noise signal is not visible or
just slightly visible on the graph.

IMPORTANT Risk of invalid data collection

or lost data if the discriminator values are
not properly set.

PN 721742AD

SOFTWARE

UNDERSTANDING THE MAIN SCREEN

7

Recover Sample button allows most of the

1%

sample that was aspirated into the instrument
to be recovered from the sample loop. You can
recover sample even if the run volume has
been completely dispensed.

IMPORTANT Misleading results can occur

due to the potential of dilution. Use care
when selecting Auto Recover Sample.

Drop down list of parameters available for

1&

collection, includes: FL1, FL2, FL3, EV, SS,
Time, FLn-FSD, FLn-FC.

Note: FLn represents either FL1, FL2 or FL3.
FSD (Fluorescence Surface Density); FC
(Fluorescence Concentration).

X-axis is the primary parameter display;

1^

triggers data collection and determines which
events are collected. There are five settings
for this button: EV, FL1, FL2, FL3, SS.

Note:

r If this graph is set to FL1, FL2, or FL3

and fluorescence is not detected during
data acquisitions, no data is collected.

r If the setting is for EV and no volume

data is detected, no data is collected.

r If the setting is for SS and no side

scatter is detected, no data is collected.

Counting Display: Displays the count rate,

1*

concentration/mL x 10
what is being collected while the sample is
running.

3

, and total count of

Parameter Information

The Parameter Information button displays a dialog box with the list of available parameters
allowing the user to customize certain fields.

Figure 7.2 Main Screen: Parameter Information

Parameter lists the available parameters for

b

collection.

Short Name allows the user to customize the

d

parameter name for buttons and labels; name
length is limited to 9 characters.

Collect checkbox allows the user to select

c

which parameters are to be collected.

Long Name is saved to the FCS listmode file

e

as $PnS keyword.

PN 721742AD

7-3

SOFTWARE

UNDERSTANDING THE MAIN SCREEN

Color drop down list allows the user to select a

f

default color assigned to the histogram data.

IMPORTANT Misleading results can occur if

the same color is assigned to parameters and
regions. Do not assign the same color to a
parameter if region has been assigned
duplicate color.

Trigger checkbox determines the primary

h

parameter.

FC checkbox determines the fluorescence

j

signal of the parameter to be used for FC data
collection.

Note: FSD or FC, not both, can be checked.

Log checkbox switches the parameter

g

between linear and logarithmic data
collection.

FSD checkbox determines the fluorescence

i

signal of the parameter to be used for FSD
data collection.

Note: FSD or FC, not both, can be checked.

Calib (Calibration) button when pressed

1)

allows calibration of FSD for antigen density
or FC for NPE and absolute concentration
fluorescence ratios, see Figure 7.3.

Calibrate FSD or FC

For detailed information regarding the FSD or FC calculated parameter, refer to Calculated

Parameters found in Chapter 4.

Figure 7.3 Main Screen: Calibrate

Calibration Type drop down list displays the

b

type of antigen density, absolute concentration,
NPE analysis.

Calibrate Using drop down list allows the user

d

to selection Region or Fixed Channel for
calibration.

Value field allows the user to input the assay

c

value for calibration.

Calibration Channel field allows the user to

e

input the channel manually for calibration.

7-4

PN 721742AD

SOFTWARE

WORKING WITH THE FILE MENU

7

Region drop down list allows the user to select

f

a region around the standard to be used for
calibration.

Cancel button returns you to the Parameter

h

Information dialog with updating the settings.

Region Statistics

Figure 7.4 Main Screen: Region Statistics

Save button saves your settings and returns

g

to the Parameter Information dialog.

Import button allows the user to import the

i

current value of the selected region to be
used for calibration

Region name can be double-clicked to edit the

b

region, see Heading 7.7, WORKING WITH THE

REGIONS MENU.

Double-clicking the desired parameter color

d

displays the Region Statistics screen, see

Heading 7.7, WORKING WITH THE REGIONS
MENU.

7.2 WORKING WITH THE FILE MENU

The File menu (Figure 5.4) allows you to save files, playback files, generate reports, export
data, and print files.

For details, see:

r Saving Listmode (.LMD) Files

r Auto Save Option

r Playing Back Files

r Defining Playback Options

r Generating a Microsoft Excel Report

r Exporting Data to Excel

Column header names can be double-clicked

c

to edit the heading label.

PN 721742AD

7-5

SOFTWARE

WORKING WITH THE FILE MENU

r Printing

r Exiting (Closing) the Application

Saving Listmode (.LMD) Files

Perform this procedure to save the data to the disk as a listmode FCS 2.0 file.

1 File tt Save.

2 Select the desired folder where you

want the file to be saved.

7-6

3 Type the file name.

4 Save.

Auto Save Option

Perform this procedure to automatically save or print results as defined in Heading 7.10,

WORKING WITH THE CURRENT SETTINGS SCREEN. When enabled, this option allows

for the LMD file, reports, Excel Files to be saved or printed and to perform flagging if Data
Flags were defined.

PN 721742AD

SOFTWARE

WORKING WITH THE FILE MENU

1 File tt Auto Save.

Note: When Auto Save is unavailable, it will appear gray in the File Menu.

2 Browse to indicate the location where the files are to be stored.

Playing Back Files

Perform this procedure to simulate running files that have previously been run. You can also
modify the file and re-save it with the new information.

7

IMPORTANT Erroneous results can occur if using FCS 2.0 files that have been generated on other systems,

other than the Quanta instruments.

1 File tt Playback.

2 Select the desired file to be played back:

a. Locate the file in the appropriate

folder.

b. Select the file.

c.

Open.

PN 721742AD

7-7

SOFTWARE

WORKING WITH THE FILE MENU

3 During file playback, a slider appears on the left of the first histogram that allows you to

control the speed of the file playback.

Note: Return to normal operation of acquiring data from the instrument by

Normal Mode.

Defining Playback Options

Perform this procedure to define playback options, which allow you to customize the
playback information. The options include:

Quick Playback allows you to view the results of a past run without having to wait through

the length of collection time.

Playback Regions, Playback Volume, and Playback Compensation Value (when selected) play

back the chosen file using the file’s original settings. If these options are not selected, the
files play back using the instrument’s current settings.

1 File tt Playback Options.

File tt

2 Select the desired playback options:

a. next to the desired option.

b. appears when the option is

selected.

Note: To deselect an option,

until appears.

3 OK to save the options.

7-8

PN 721742AD

SOFTWARE

WORKING WITH THE FILE MENU

Generating a Microsoft Excel Report

Perform this procedure to generate an Excel report based on the currently selected Template
file, see Heading 7.10, WORKING WITH THE CURRENT SETTINGS SCREEN. If no
Template is selected in the protocol, then one of the Standard Template files will be used
based on the number of parameters configured, or you can create a customer Template file.

1 File tt Excel Report. to open the currently selected Excel template populated with the

active File Information.

Exporting Data to Excel

Perform this procedure to send raw listmode data to an Excel file.

7

1 File tt Export Data to Excel.

Printing

Perform this procedure to print. If you want file information printed along with the data.

1 File tt Print.

PN 721742AD

7-9

SOFTWARE

WORKING WITH THE FILE MENU

2 Select the desired printer and print

settings.

3 Print.

7-10

PN 721742AD

WORKING WITH THE INSTRUMENT MENU

Exiting (Closing) the Application

Perform this procedure to close the application, which automatically sends a Valves Off
command to the instrument.

1 File tt Exit.

2 Your Windows desktop appears after

the Quanta software is closed.

SOFTWARE

7

7.3 WORKING WITH THE INSTRUMENT MENU

The Instrument menu (Figure 5.5) allows you to do the following procedures. This menu is
disabled after playing back a listmode file; select

r Start Up

r Shut Down

r Defining the Power Setting

r Running the Cleaning Cycle

r Flushing

r Fill Cup

r Resetting Fluid Count

r Setting Up the Instrument

r Importing Settings

r Automatically Aligning the Optics

File tt Normal Mode to end Playback Mode.

PN 721742AD

7-11

SOFTWARE

WORKING WITH THE INSTRUMENT MENU

Start Up

Perform this procedure to do a Start Up routine on the instrument.

1 Place an empty sample cup on the

instrument.

2 Instrument tt Start Up.

3 Follow the instructions on the screen.

Refer to Heading 6.1, PERFORM

STARTUP for detailed instructions.

7-12

PN 721742AD

WORKING WITH THE INSTRUMENT MENU

4 Next Step. The screen will prompt you through the proper procedure to startup the

instrument.

IMPORTANT Erroneous results can occur if both light systems are active at the same time. Ensure that the

appropriate light source is selected during the startup procedure, refer to Heading 6.1, PERFORM

STARTUP for detailed instructions.

Shut Down

Perform this procedure run a Shut Down routine on the instrument.

1 Instrument tt Shut Down.

SOFTWARE

7

2 Follow the instructions on the screen.

Refer to Heading 6.3, PERFORM

SHUTDOWN for detailed instructions.

3 Next Step. The screen will prompt you through the proper procedure to shutdown the

instrument, see Heading 6.3, PERFORM SHUTDOWN.

PN 721742AD

7-13

SOFTWARE

WORKING WITH THE INSTRUMENT MENU

Defining the Power Setting

After the lamp has completed its initial warm up cycle of about 20 minutes, perform this
procedure once weekly to define the correct setting for the arc lamp potentiometer located on
the power supply next to the voltage display.

1 Instrument tt Power Setting.

2 Open the cover.

C

3 Obtain the current voltage from the

power supply inside the instrument.
Voltage reading should be between
19-28 volts.

B

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7-14

PN 721742AD

4 Type the lamp voltage; notice that the

Potentiometer Setting automatically
changes.

5 Set the Potentiometer to the new

Potentiometer setting value.

SOFTWARE

WORKING WITH THE INSTRUMENT MENU

7

PN 721742AD

6 Record the lamp potentiometer setting in the instrument logbook per your laboratory

requirements.

7-15

SOFTWARE

WORKING WITH THE INSTRUMENT MENU

Running the Cleaning Cycle

Perform this procedure to remove flow cell clogs. The system runs the cleaning solution
through the aperture at a high speed.

1 Place an empty sample cup on the

instrument.

2 Instrument tt Cleaning Cycle. The

following screen displays. Follow the
directions on the screen to continue
with the Cleaning Procedure.

7-16

PN 721742AD

3 Next Step. The following screen

displays. Follow the directions on the
screen to continue with the Cleaning
Procedure.

SOFTWARE

WORKING WITH THE INSTRUMENT MENU

7

Flushing

Perform this procedure to empty fluid from the electrodes and certain areas of the flow cell
and to refill the locations. The system does this by running sheath fluid through the flow cell
at a high speed.

1 Place an empty sample cup on the

instrument.

PN 721742AD

7-17

SOFTWARE

WORKING WITH THE INSTRUMENT MENU

2 Instrument tt Flush (F).

Fill Cup

Perform this procedure to fill the sample cup with diluent.

1 Place an empty sample cup on the

instrument.

7-18

2 Instrument tt Fill Cup.

Resetting Fluid Count

Perform this procedure to reset the fluid count after the sheath bottle has been emptied and
re-filled or after the waste container has been emptied to ensure proper level sensor
monitoring.

1 Instrument tt Reset Fluid Count.

PN 721742AD

WORKING WITH THE INSTRUMENT MENU

2 OK.

Setting Up the Instrument

Only your Beckman Coulter Representative can perform this procedure to define the
instrument settings. For additional information on the Instrument Setup screen, see

Understanding the Instrument Setup Screen.

1 Instrument tt Instrument Setup.

SOFTWARE

7

2 This screen is displayed for reference

information only. The user cannot
modify any settings which appear on
this screen display.

PN 721742AD

7-19

SOFTWARE

WORKING WITH THE INSTRUMENT MENU

Understanding the Instrument Setup Screen

See Figure 7.5.

Figure 7.5 Instrument Setup Screen: Defined

7-20

b

d

f

h

Identifies the instrument model

Defines the maximum number of
parameters that the instrument can collect

Defines the sample volume (about

54.5 µL) that must be boosted from the
sample loop to the flow cell.

Set Boost Size button allows you to
automatically configure the boost size.

Defines the sample volume (about

57.0 µL) from the aspirator tip to the first
T, where valve 7 is located

c

e

g

i

Identifies the serial number on back of the
instrument

Identifies the valve scenario (e.g. Pump)
used by the instrument

Diagnostics button displays the
Diagnostics screen.

Defines the comm port on the PC where
the metering pump is connected

PN 721742AD

SOFTWARE

WORKING WITH THE INSTRUMENT MENU

7

j

1!

1#

1%

1&

1(

Defines the syringe size

Indicates (when checked) that an arc lamp
is present on the instrument

Defines the com port on the PC where the
laser is connected

Cancels unsaved settings.

Saves settings.

ADC Setup box contains configurable
parameter name, detector type and on/off
checkbox. These options are available to
Service only.

1)

1@

1$

1^

1*

Defines the aperture size

Indicates (when checked) that a laser is
present on the instrument

Collect checkbox turns ADC on or off.
These options are available to Service
only.

Detector Type drop down list provides the
detection options for each parameter to be
collected. These options are available to
Service only.

Parameter Name drop down list allows the
selection of the parameter to be used for
collection. These options are available to
Service only.

PN 721742AD

7-21

SOFTWARE

WORKING WITH THE INSTRUMENT MENU

Understanding the Diagnostic Functions Screen

See Figure 7.6.

Figure 7.6 Diagnostic Functions Screen: Defined

7-22

CAUTION Instrument damage can occur if diagnostics functions are attempted by the user without

supervision of a Beckman Coulter Representative. Do not attempt to perform these Diagnostic Functions
without direction from a Beckman Coulter Representative.

b

d

f

This section is used to activate different
fluidics functionality.

This section is used to activate, control
and manage the syringe pump
performance.

This section is used to individually verify
functionality of the valves or different
programmed valve scenarios.

c

e

g

This section is used to individually verify
functionality of the valves or different
programmed valve scenarios.

This section is used to activate, control
and manage the syringe pump
performance.

This section is used to activate the four
stepping motors for Auto Alignment and
Lamp functions.

PN 721742AD

Laser Control

Perform this procedure to adjust the laser power.

1 Instrument tt Laser Control.

Understanding the Laser Control Options

See Figure 7.7.

Figure 7.7 Laser Control Options: Defined

SOFTWARE

WORKING WITH THE INSTRUMENT MENU

7

PN 721742AD

b

d

f

h

Scroll bar adjusts the laser power to a
lower value to optimize CVs when the
sample is extremely bright and is flooding
the detectors.

Note: Laser Power is normally set to 22
mW.

Displays the temperature of the diode.

Cycle checkbox when enabled turns the
laser ON at the start of acquisition and OFF
when acquisition stops.

If unchecked, laser remains ON at all
times.

On button turns the laser ON.

c

e

g

i

Displays the container temperature.

Displays the laser current.

Off button turns the laser OFF.
Note: If Cycle checkbox not enabled, turn

OFF the laser when not using the
instrument.

Laser Power displayed in mW.

7-23

SOFTWARE

WORKING WITH THE INSTRUMENT MENU

Importing Settings

Perform this procedure to import instrument settings and protocols from other software
versions transferred to your instrument. For additional information, see Understanding the

Import Settings Screen.

1 Instrument tt Import Settings.

2 Select the desired options:

a. to browse your computer

for the desired database file
(Quanta.mdb).

b. next to the desired option.

c. appears when the option is

selected.

Note: To deselect an option,

until appears.

3 Import.

a. When the import is complete, an Import Complete message appears.

b.

OK to return to the main screen.

7-24

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Understanding the Import Settings Screen

See Figure 7.8.

Figure 7.8 Import Setting Screen: Defined

SOFTWARE

WORKING WITH THE INSTRUMENT MENU

7

b

d

f

h

Version/File from which to import

Browse to pick a file rather than type in a
name

Button to perform the import

Import protocols yes/no

c

e

g

i

Installed versions from which to select

Cancels unsaved settings.

Replace existing protocols with the same
name

Import instrument settings yes/no

Automatically Aligning the Optics

Note: This menu option is only available when running a sample.

Perform this procedure if your HPCVs are still not within specifications. For detailed
instructions, refer to Heading 8.5, AUTOMATICALLY ALIGNING THE OPTICS.

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7-25

SOFTWARE

WORKING WITH THE INSTRUMENT MENU

Understanding the Auto Optical Alignment Screen

See Figure 7.9.

Figure 7.9 Auto Optical Alignment Screen: Defined

7-26

b

d

f

h

j

Align Using drop down list selects the
fluorescence parameter that corresponds
to the beads being run.

Focus checkbox when enabled adjusts the
microscope focus when Align button is
selected.

Lamp X axis checkbox when enabled
adjusts the arc lamp along the X-axis when
the Align button is selected.

Align button begins the automatic
alignment.

Graph displays the Peak Channel, CV and
Alignment value at each motor position
during alignment.

c

e

g

i

Count Rate displays the count rate.

Position checkbox when enabled adjusts
the microscope position when Align
button is selected.

Lamp Y axis checkbox when enabled
adjusts the arc lamp along the Y-axis when
the Align button is selected.

Exit button closes the Optical Alignment
window.

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7.4 WORKING WITH THE GAIN MENU

For additional information, see Understanding the Tracking Settings Screen.

Tracking is the auto gain adjustment done by the computer to keep a known sample in the
same channel number. As the Mercury Arc lamp ages, its intensity increases. The effects of the
arc lamp’s aging are usually nominal, but tracking may be necessary on runs where maximum
resolution is needed to separate close populations.

The Gain menu (Figure 5.6) allows you to do the following procedures:

r Show Gain Settings

r Defining Tracking Settings

r Tracking Start

r Reset Tracking

Show Gain Settings

Perform this procedure to display the Gain Settings on the Main Screen of each fluorescent
parameter and PMT voltage of the fluorescence parameters.

SOFTWARE

WORKING WITH THE GAIN MENU

7

1 Gain tt Show Gain Settings.

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7-27

SOFTWARE

WORKING WITH THE GAIN MENU

Defining Tracking Settings

Perform this procedure to define the tracking settings.

1 Gain tt Tracking Settings.

2 Select the specific parameter tab to

modify the tracking settings.

a. the Enable Tracking box.

b. Enter the Narrow Tracking Width

and Wide Tracking Width.

c.

Save to close dialog box and

save your new settings or

Cancel to discard the updated

settings.

3 To begin Tracking, Gain tt Tracking Start.

4 To stop Tracking, Gain tt Tracking Stop.

7-28

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Understanding the Tracking Settings Screen

See Figure 7.10.

Figure 7.10 Tracking Settings Screen: Defined

SOFTWARE

WORKING WITH THE GAIN MENU

7

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b

d

f

h

Parameter tabs appear in order defined by
the user depending upon the trigger
parameter and order set.

Enable checkbox to clear the data after the
standard is first locked to the correct
channel

Set to the width of the standard peak

Channel in which the standard should
reside

c

e

g

i

Select to cancel your setting changes.

Select to save your setting changes

Set to a value approximately equal to
one-half of the standard peak width

Enable checkbox to track the channel on
this parameter tab

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SOFTWARE

WORKING WITH THE GAIN MENU

Tracking Start

Tracking is the auto gain adjustment performed by the computer to keep a known sample
within the same channel number. This adjustment may become necessary as an arc lamp ages
and its intensity increases. The effects of arc lamp aging is not usually large; however, tracking
may be required for maximum resolution on long sample runs.

Perform this procedure to start tracking. The button toggles between Start and Stop. It
controls if tracking is ON or OFF.

1 Gain tt Tracking Start.

2 The Tracking Display on the Main

screen overlays the Region Display. By
default, when tracking is enabled, the
Tracking Display is set to visible.

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WORKING WITH THE VOLUME MENU

Reset Tracking

Perform this procedure to restore the gain settings to their original values before tracking
started.

1 Gain tt Tracking Reset.

2 The Tracking Display on the Main

screen is reset when this selection is
pressed.

SOFTWARE

7

7.5 WORKING WITH THE VOLUME MENU

The Volume menu (Vol um e M en u) allows you to calibrate the volume and display channels.
For details, see:

r Calibrating the Volume

r Displaying Channels

p

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Calibrating the Volume

Perform this procedure to calibrate the EV axis of the data graphs and the mean cell volume,
diameter and surface area statistics to absolute units. To ensure proper calibration, use the
correct size of Coulter Calibration beads. For additional information, see Understanding the

Volume Calibration Screen.

7-31

SOFTWARE

WORKING WITH THE VOLUME MENU

Calibration Beads

Dilute the Coulter Calibration beads as follows:

Bead µm Part Number Dilution Ratio

1 µm 6602790 1:2000

2 µm 6602792 1:5

3 µm 6602793 1:5

5 µm 6602794 1:3

10 µm 6602796 1:2

20 µm 6602798 1:1

1 Place 1 mL of beads in a sample cup.

a. Mix gently and place onto sample

holder.

b. This sample will be used to set up

the single parameter gate region.

7-32

2 Start.

3 Adjust the EV gain to place the beads in Channel 200 on the EV graph. Refer to

Heading 7.4, WORKING WITH THE GAIN MENU for detailed instructions.

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WORKING WITH THE VOLUME MENU

4 Adjust the lower discriminator to eliminate the noise. Refer to Heading 7.1,

UNDERSTANDING THE MAIN SCREEN for detailed instructions.

5 Go to the Regions Menu and set up a

Single Parameter Region. Refer to

Managing Regions for detailed

instructions on setting up this region.

SOFTWARE

7

6 Collect at least 5000 data points.

a.

Stop to stop the sample.

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7-33

SOFTWARE

WORKING WITH THE VOLUME MENU

7 Volume tt Calibrate.

a. Select

Diameter from the Calibrate

For

and Calibrate Using drop down

lists.

b. Insert the diameter assayed value

of the beads in the drop down list
to the right of

Calibrate using list.

Refer to the COULTER CC Size
Standard reagent package insert
for the assay values.

c.

Import and select the region

around the beads. The Mean
Channel of the region should
appear in the box labeled

Number.

Channel

8 Calibrate.

Note: Volume automatically recalibrates if the EV gain is adjusted after calibration.

7-34

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