BD FACSymphony A5 SE User manual

BD FACSymphony™ A5SE
Flow Cytometer
User's Guide
23-23473(02)
2022-08
English
For Research Use Only

Copyrights
No part of this publication may be reproduced, transmitted, transcribed, stored in retrieval systems, or translated into any language or
computer language, in any form or by any means: electronic, mechanical, magnetic, optical, chemical, manual, or otherwise, without prior
written permission from BD.
The information in this guide is subject to change without notice. BD reserves the right to change its products and services at any time.
Although this guide has been prepared with every precaution to ensure accuracy, BD assumes no liability for any errors or omissions, nor for
any damages resulting from the application or use of this information. BD welcomes customer input on corrections and suggestions for
improvement.
Trademarks and patents
BD, the BD Logo, BDFACSClean, BDFACSDiva, BDFACSFlow, BDFACSymphony, BDHorizonBrilliant, BDHorizon RealYellow, FACS and
Horizon are trademarks of Becton, Dickinson and Company or its affiliates. All other trademarks are the property of their respective owners.
© 2022 BD. All rights reserved.
For US patents that may apply, see bd.com/patents.
Alexa Fluor™ is a trademark of Life Technologies Corporation.
CF™ is a trademark of Biotium, Inc.
Cy™ is a trademark of GE Healthcare. Cy™ dyes are subject to proprietary rights of GE Healthcare and Carnegie Mellon University, and are
made and sold under license from GE Healthcare only for research and in vitro diagnostic use. Any other use requires a commercial
sublicense from GE Healthcare, 800 Centennial Avenue, Piscataway, NJ 08855-1327, USA.
Regulatory information
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
Laser safety information
Class 1 Laser Product.
FCC information
WARNING: Changes or modifications to this unit not expressly approved by the party responsible for compliance could void the user’s
authority to operate the equipment.
NOTICE: This equipment has been tested and found to comply with the limits for a Class A digital device, pursuant to Part 15 of the FCC
Rules. These limits are designed to provide reasonable protection against harmful interference when the equipment is operated in a
commercial environment. This equipment generates, uses, and can radiate radio frequency energy and, if not installed and used in
accordance with the instruction manual, may cause harmful interference to radio communications. Operation of this equipment in a
residential area is likely to cause harmful interference in which case the user will be required to correct the interference at his or her own
expense. Shielded cables must be used with this unit to ensure compliance with the Class A FCC limits. This Class A digital apparatus meets
all requirements of the Canadian Interference-Causing Equipment Regulations. Cet appareil numérique de la classe A respecte toutes les
exigences du Réglement sur le matériel brouilleur du Canada.
History
Revision Date Change made
23-23473-00 2021-07 Initial release
23-23473(01) 2021-12 In topic Spectral function overview (page 48), corrected UVchannel A filter
bandpass value. Extended list of commercial dyes and the appropriate laser
detectors to use with them.
Changed laser delay ordering and associated delay-optimization procedure.
23-23473(02) 2022-08 Updated UVlaser information to indicate that the laser emission frequency
can be 349nm or 355nm.
Updated table of detectors used with common dyes in Spectral function
overview (page 48)
Updated screenshots to reflect user interface enhancements.

Contents
BD FACSYMPHONY™ A5SE 1
1 ABOUT THIS GUIDE 7
What this guide covers 8
Conventions 8
About the documentation 9
Instrument technical support 10
2 INTRODUCTION 11
System overview 12
Cytometer overview 12
Control panel 14
Fluidics system 15
Sheath and waste containers 19
Optics 20
Workstation 22
3 CYTOMETER SETUP 23
Starting the cytometer and computer 24
Preparing the sheath container 25
Removing air bubbles 27
Preparing the waste container 29
Preparing the fluidics 31
Adding a bead lot number 32
Custom configurations and baselines 34
4 MAINTENANCE 35
Maintenance overview 36
Cleaning the fluidics 37
Shutting down the cytometer 38
Flushing the system 38

Replacing the waste container cap 40
Changing the sheath filter 41
Changing the Bal seal 42
Changing the sample tube O-ring 44
Cleaning or replacing the sheath gasket 45
5 SPECTRAL FUNCTIONS 47
Spectral function overview 48
Autofluorescence 54
Spectral unmixing software considerations 55
Setting up a new spectral experiment 57
Creating a Spectral Unmixing matrix 59
Recording and analyzing spectral data 62
Repeating a spectral experiment 65
6 OPTIMIZING CYTOMETER SETTINGS 67
Cytometer settings workflow 68
Verifying the configuration and user preferences 69
Running a performance check 71
Setting up a compensation experiment 74
Creating application settings 77
Recording compensation controls 78
Calculating compensation 81
7 RECORDING AND ANALYZING DATA 83
Data recording and analysis workflow 84
Preparing the workspace 84
Recording data 85
Analyzing data 87
Reusing an analysis 91
8 MANUAL SETTINGS 93
About laser delay 94
Optimizing laser delay 95
Adjusting area scaling 97
9 TROUBLESHOOTING 103
Cytometer troubleshooting 104
Electronics troubleshooting 109
10 SUPPLIES AND CONSUMABLES 111
Ordering information 112
Beads 112
Reagents 113
4BD FACSymphony™ A5 SE User's Guide

Equipment 113
INDEX 115
Contents 5

BD FACSymphony™ A5 SE User's Guide

11
About this guide
This chapter covers the following topics:
lWhat this guide covers (page 8)
lConventions (page 8)
lAbout the documentation (page 9)
lInstrument technical support (page 10)

What this guide covers
This guide describes the procedures necessary to operate and maintain the BDFACSymphony™A5SE Special
Order Research Product (SORP) flow cytometer.
Because many cytometer functions are controlled by BDFACSDiva™ software, this guide also contains
information about software features required for basic cytometer setup and operation.
This guide assumes you have a working knowledge of basic Microsoft® Windows® operation. If you are not
familiar with the Windows operating system, see the documentation provided with your computer.
Conventions
Introduction
The following table lists the safety symbols used in this guide to alert you to potential hazards.
Safety symbols
Symbol1Description
Caution alert.
Identifies a hazard or unsafe practice that could result in data loss, material damage, minor injury, severe
injury, or death.
Electrical hazard
Biological hazard
Laser hazard
8BD FACSymphony™ A5 SE User's Guide

About the documentation
Introduction
This topic describes the documentation available with the BDFACSymphony™A5SE flow cytometer.
The following list includes the available documentation for the system.
lBDFACSDiva™ Software Reference Manual: Includes instructions or descriptions for installation and setup,
workspace components, acquisition controls, analysis tools, and data management. Access this manual from
the BDFACSDiva™ Software Help menu (Help > Documentation > Reference Manual), or by double-clicking
the shortcut on the desktop.
lBD®Cytometer Setup and Tracking Application Guide: Describes how to use the BD®Cytometer Setup and
Tracking (CS&T) features in BDFACSDiva™ software.
lBDFACSymphony™A5SE Flow Cytometer Site Preparation Guide contains specifications for:
oCytometer weight and size
oTemperature and other environmental requirements
oElectrical requirements
lBDFACSymphony™A5SE Flow Cytometer Safety and Limitations: Provides descriptions of safety and
warning labels, general system hazards, specific risks, and laser, electrical, and biological hazards.
lBD®High Throughput Sampler User’s Guide: Describes how to set up and operate the BD®High Throughput
Sampler (HTS) option. It also contains a description of BDFACSDiva™ software features specific to the HTS.
lBDFACSFlow™ Supply System User’s Guide: Describes the optional automated sheath and waste fluid control
system.
Publication formats
This guide is provided in PDF format to provide an eco-friendly option.
Chapter 1 About this guide 9

Instrument technical support
Introduction
This topic describes how to get technical assistance.
Contacting technical support
If technical assistance is required, contact your local BDBiosciences customer support representative or
supplier. Go to our website bdbiosciences.com for up-to-date contact information.
When contacting BD Biosciences, have the following information available:
lProduct name, part number, and serial number
lVersion of BDFACSDiva™ software you are using
lAny error messages
lDetails of recent system performance
More information
lWhat this guide covers (page 8)
10 BD FACSymphony™ A5 SE User's Guide