Flow Cytometry

Flow Cytometer Buyer’s Guide
From Instrument to Configuration

Visit bio-rad.com/cellanalysis for more information.

What to Consider When Buying a Flow Cytometer

A flow cytometer is a significant investment for your laboratory and there are many
considerations to ensure that you choose an appropriate instrument to meet yourusers’ needs. Things like the number of parameters it can measure, the throughput
requirements, and sample handling will all need to be factored into your decision.
This Buyer’s Guide highlights some key considerations to enable you to narrow
down your instrument and configuration options.

2 | BIO-RAD LABORATORIES, INC.

Define Your Needs
Having a clear idea of what your users’ needs are will pay huge dividends. Not only will it guarantee that you approach your purchasein a practical way, but it will help you justify your budget for new equipment. If you are in a multiuser environment, consider identifying
gaps in the current service by surveying your userbase. As a flow cytometer is a substantial investment, make sure you think long-term;
anticipating your future needs is almost as important as the here and now.
The Bio-Rad ZE5 Cell Analyzer is designed with flexibility in mind to help adapt to future use. Its integrated plate loader can handle 1.5 and
5 ml tubes as well as plates, and it can be upgraded with the addition of extra lasers and detectors if your needs change. It is automationready and ideal for high-throughput screening and routine laboratory applications.
Prioritize Your Applications
Anticipate the different types of applications that your flow cytometer will be routinely used for so you can identify essential features andprioritize laser combinations. For example, if immunophenotyping is a priority, the inclusion of violet (405 nm) and ultraviolet (UV; 355 nm)
lasers will help expand the number of compatible fluorophores and enables more flexible panel design.
The ZE5 Cell Analyzer offers a host of features that ensure it is suitable for a wide range of applications, including immunophenotyping,exosome detection, fluorescence resonance energy transfer (FRET), bead arrays, and high-throughput screening. The system can bepreconfigured with different laser combinations based on your priority applications (Table 1).
Table 1. Laser priority by application. Larger circles represent higher priority.
Application

Laser
355 nm*Immunophenotyping

405 nm488 nmIncluded

Small particles**FRET

640 nmExplanation
UV* and violet lasers provide the highest number of colors.

Included

The ZE5 Cell Analyzer can be equipped with a small particle
detector triggered off the violet laser. The brightness ofphycoerythrin (PE), which is maximally excited by the yellow(561 nm) laser, is also useful for small particle detection.

Included

The yellow and violet lasers give the best choice of compatible
FRET pairs.

IncludedGenerally, markers with only a few conjugates will be available in
only fluorescein isothiocyanate (FITC), PE, and allophycocyanin(APC). The blue laser (488 nm) covers FITC and PE and the redlaser APC.

Included

The red laser is often needed for bead arrays and the violet laser
gives maximum flexibility for other applications.

Novel markers

Bead assays

561 nm* Only available for 4 and 5 laser configurations.
** Also consider the small particle detector.
APC, allophycocyanin; FITC, fluorescein isothiocyanate; FRET, fluorescence resonance energy transfer; PE, phycoerythrin; UV, ultraviolet.

The small particle detector is an additional scatter detector triggered from the violet laser and calibrated to give enhanced resolutionof small particles such as bacteria, yeast, and extracellular vesicles. It is included as standard on all five-laser ZE5 Cell Analyzer
configurations. Four- and three-laser configurations that have a violet laser are also available with a small particle detector. It is also anoption as a field upgrade for four- and three-laser systems equipped with a violet laser.
Count Your Parameters
How many parameters do you need to measure? The number will determine which laser configuration is best for you:
For 17 or fewer, consider a three-laser configuration
For 17–24, consider a four-laser configuration■ For 24–27, consider a five-laser configuration that includes a small particle detector■■For complex immunophenotyping assays, think about expanding the number of parameters available off the UV laser from five to seven.
Note that, in this configuration, yellow laser parameters are reduced from seven to five. Refer to the ZE5 Cell Analyzer Laser and Filter
Configuration Guide on pages 6–7 when configuring your ZE5 Cell Analyzer.

BIO-RAD LABORATORIES, INC. | 3

Think About Throughput
Ensure that your instrument can handle the volume of samples that you need to process for your current workload, but also factor in a
50% increase in throughput to ensure it can also meet your needs if they evolve. Avoid having to double up to meet throughput demands
by choosing a flow cytometer that has additional capabilities, ready for future applications and experiments.
The Bio-Rad ZE5 Cell Analyzer can collect data at a higher rate than any other instrument currently available. The electronics and fluidics
make it capable of sampling data at 100,000 events per second without compromising data quality, making rare event detection superfast. Additionally, in high-throughput mode plates can be processed at speeds previously only available with instruments dedicated toscreening: a 96 well plate may take less than 15 minutes and a 384 well plate less than 50 minutes. Combining these two advantages
makes it possible to perform complex immunophenotyping panels quickly and maximize productivity.
Consider Automation
The ability to screen more and analyze faster while maintaining high quality, accurate results is key to progressing research. Automating
your flow cytometry workflow can dramatically increase productivity and lead to greater progress. As not all instruments are suitable
for automation, future proof your workflow by making sure that you opt for an instrument that was designed with automation in mind.
Robotic integration and automation features simplify operations, save time, and improve lab performance. Automation providers will behappy to offer further assistance with setting up your workstation.

External Fluidics are especially useful if you are considering automation. The house deionized (DI) connection allows the ZE5 Cell Analyzer to be connected directly to a supplyof DI water. The external bulk carboys replace the internal tanks to give up to 24 hr uninterrupted run time.

Automation-ready, the ZE5 Cell Analyzer has been integrated by many providers into a wide range of automation workcells.
Large capacity fluidics options ensure uninterrupted 24 hour operation.
Visit bio-rad.com/htscreening for more information on ZE5 Cell Analyzer automation.
Identify Your Installation Site
Laboratory space is often at a premium, especially in more established labs. Think about space below, as well as above, the benchand don’t forget to factor in access for servicing. If considering automation, the flexibility of modern robotics allows for workcells to
be built in more limited spaces than previously so automation can be possible even in smaller spaces. It’s a good idea to contact yourlocal instrument representative as well as your automation provider to seek advice if space is limited.
Unlike many other systems, the ZE5 Cell Analyzer does not need a separate fluidics cart, all the fluidics can be contained within
the instrument itself, although larger external tanks are available as well. With a remarkably small footprint, it can be installed on astandard benchtop.

4 | BIO-RAD LABORATORIES, INC.

Next Steps
Make a Shortlist and Use Your Network
Once you have a clear idea of what you need, make a shortlist of instruments that match your requirements. Then use your network
to gain experience from your peers. Speak to your colleagues as well as the wider userbase on social media, at meetings, and atconferences to get a broad opinion of the instruments on your list.
Talk with a Specialist
Contact your local sales teams and speak with a specialist in flow cytometry within the company. They should ensure that the instrument
you have selected meets your requirements and is the right configuration for you. This is your chance to obtain more detailed information,so make sure that you have a list of questions as well as providing as much detail as possible on what you need from the instrument.
For a neutral appraisal of the technology, talk to other researchers using the same system you are considering.
Request a Demo
Once you have identified a flow cytometer that meets your requirements, put it through its paces by arranging a demonstration.
Involve key stakeholders in the demonstration and perform your most important assays to ensure the machine will deliver what
you hope. Plan to have suitable samples ready for your demonstration.

BIO-RAD LABORATORIES, INC. | 5ZE5 Cell Analyzer Laser and Filter Configuration Guide

5-Laser (27 colors)
Laser

355

FilterFluorochrome

SBUV400, BUV395

387/11

SBUV400, BUV395

387/11

SBUV400, BUV395

447/60

SBUV445, AF350, DAPI

509/24

SBUV510, DAPI, Zombie UV

460/22525/50

SBUV510, BUV496

577/15

SBUV575, BUV536

SBUV445, DAPI, Zombie UV,
L/D Blue

670/30

SBUV665, BUV661

615/24

SBUV605, BUV615

509/24

SBUV510, BUV496

670/30

SBUV665, BUV661

577/15

SBUV575, BUV563

747/33

SBUV740, BUV737

670/30

SBUV665, BUV661

780 LP

SBUV795, BUV805

747/33

SBUV740, BUV737

420/10

BV421

780 LP

SBUV795, BUV805

460/22

SBV440, Pacific Blue

420/10BV421

525/50

SBV515, BV510, CFP

460/22

SBV440, Pacific Blue

615/24

SBV610, BV605

525/50

SBV515, BV510, CFP

670/30

SBV670, BV650

615/24

SBV610, BV605

720/60

SBV710, BV711

670/30

SBV670, BV650

750LP

SBV760, SBV790, BV786

720/60

SBV710, BV711

488/10Side Scatter

750LP

SBV760, SBV790, BV786

525/35

FITC, GFP, YFP, AF488,
Kiravia520

488/10Side Scatter

525/35

FITC, GFP, YFP, AF488,
Kiravia520

387/11

488

561

640

Laser

5-Laser 7 Off UV Option B (27 Colors)

Fluorochrome

Fluorochrome

700LP

405

5-Laser 7 Off UV Option A (27 Colors)
FilterFilter355

SBUV740, SBUV795,
BUV737420/10BV421

460/22

SBV440, Pacific Blue

525/50

SBV515, BV510, CFP

615/24

SBV610, BV605

670/30

SBV670, BV650

720/60

SBV710, BV711

750LP

SBV760, SBV790, BV786

488/10Side Scatter

525/25

FITC, GFP, YFP, AF488,
Kiravia520

593/52

SBB580692/80

SBB675, SBB700,
PerCP-Cy5.5

750LP

SBB765, SBB810

577/15

SBY575, PE

589/15

DsRed, tdTomato

615/24

SBY605, PE-Dazzle,
PE-CF594, mCherry

640/20mPlum

670/30

SBY665, PE-Cy5, PE-AF647

720/60

SBY720, PE-Cy5.5

405

488

561

593/52

SBB580692/80

SBB675, SBB700,
PerCP-Cy5.5

355

405

488

593/52

SBB580692/80

SBB675, SBB700,
PerCP-Cy5.5

750LP

SBB765, SBB810

583/30

SBY575, PE, DsRed,
tdTomato, RFP

750LP

SBB765, SBB810

583/30615/24

SBY605, PE-Dazzle,
PE-CF594, mCherry

SBY575, PE, DsRed,
tdTomato, RFP

615/24670/30

SBY665, PE-Cy5,PE-AF647

SBY605, PE-Dazzle,
PE-CF594, mCherry

670/30

SBY720, PE-Cy5.5

SBY665, PE-Cy5,
PE-AF647

720/60

SBY720, PE-Cy5.5

750LP

SBY800, PE-Cy7, PE-AF750

670/30APC, AF647

720/60AF700

775/50APC-Cy7, AF750

800LP

AF790, APC/Fire810

750LP

SBY800, PE-Cy7, PE-AF750

720/60670/30APC, AF647

750LP

SBY800, PE-Cy7, PE-AF750

720/60AF700

670/30APC, AF647

775/50APC-Cy7, AF750

720/60

AF700

800LP

AF790, APC/Fire810

775/50APC-Cy7, AF750

800LP

AF790, APC/Fire810

640

Laser

561

640

Note: This is not a comprehensive list of fluorophores and dyes that can be used with the ZE5 Cell Analyzer.

continues

6 | BIO-RAD LABORATORIES, INC.

ZE5 Cell Analyzer Laser and Filter Configuration Guide, continued

4-Laser (24 colors)*,**
Laser

405

488

561

640

FilterFluorochrome

420/103-Laser (17 Colors)*,**
Laser

FilterFluorochrome

BV421

420/10BV421

460/22

SBV440, Pacific Blue

460/22

SBV440, Pacific Blue

525/50

SBV515, BV510, CFP

525/50

SBV515, BV510, CFP

615/24

SBV610, BV605

615/24

SBV610, BV605

670/30

SBV670, BV650

670/30

SBV670, BV650

720/60

SBV710, BV711

720/60

SBV710, BV711

750LP

SBV760, SBV790, BV786

750LP

SBV760, SBV790, BV786

488/10Side Scatter488/10509/24

FITC, GFP, AF488,
Kiravia520

509/24405

3-Laser Option 2 (17 colors)**
Laser

FilterFluorochrome

Side Scatter488/10Side Scatter

FITC, GFP, AF488,
Kiravia520

509/24

FITC, GFP, AF488,
Kiravia520549/15

YFP

549/15

YFP

549/15

YFP

583/30

SBB580, PE

583/30

SBB580, PE

583/30

SBB580, PE

615/24

SBB615, PE-CF594,
PE-Dazzle

615/24

SBB615, PE-CF594,
PE-Dazzle

615/24

SBB615, PE-CF594,
PE-Dazzle

692/80

SBB675, SBB700,
PerCP-Cy5.5, PE-Cy5692/80

SBB675, SBB700,
PerCP-Cy5.5, PE-Cy5692/80

SBB675, SBB700,
PerCP-Cy5.5, PE-Cy5

750LP

SBB765, SBB810, PE-Cy7,
PE-AF750

750LP

SBB765, SBB810, PE-Cy7,
PE-AF750

750LP

SBB765, SBB810, PE-Cy7,
PE-AF750

488488577/15

SBY575, PE

577/15

SBY575, PE

589/15

DsRed, tdTomato

589/15

DsRed, tdTomato

615/24

SBY605, PE-Dazzle,
PE-CF594, mCherry

615/24

SBY605, PE-Dazzle,
PE-CF594, mCherry

640/20mPlum

640/20mPlum

670/30

SBY665, PE-Cy5, PE-AF647

670/30

SBY665, PE-Cy5, PE-AF647

720/60

SBY720, PE-Cy5.5

720/60

SBY720, PE-Cy5.5

750LP

SBY800, PE-Cy7, PE-AF750

750LP

SBY800, PE-Cy7, PE-AF750

670/30APC, AF647

670/30APC, AF647

670/30APC, AF647

720/60AF700

720/60AF700

720/60AF700775/50

APC-Cy7, AF750

775/50APC-Cy7, AF750

775/50APC-Cy7, AF750

800LP

AF790, APC/Fire810

800LP

AF790, APC/Fire810

800LP

AF790, APC/Fire810

561

640

640

3-Laser (20 Colors)*,**
Laser

FilterFluorochrome

420/10BV421

460/22

SBV440, Pacific Blue,
mTurquoise525/50

SBV515, BV510, CFP,
Cascade Yellow

405

488

561

615/24

SBV610, BV605

670/30

SBV670, BV650

720/60

SBV710, BV711

750LP

SBV760, SBV790, BV786

488/10Side Scatter

509/24

FITC, eGFP, AF488,Kiravia520, mClover

549/15eYFP

583/30SBB580, PE

615/24

SBB615, PE-CF594,PE-Dazzle

692/80

SBB675, SBB700,
PerCP-Cy5.5, PE-Cy5

750LP

SBB765, SBB810, PE-Cy7,PE-AF750

577/15

SBY 575, PE

589/15

DsRed, tdTomato, RFP

615/24

SBY605, PE-Dazzle,
PE-CF594, mCherry

640/20mPlum

670/30

SBY665, PE-Cy5, PE-AF647

720/60

SBY720, PE-Cy5.5

750LP

SBY800, PE-Cy7, PE-AF750

Note: This is not a comprehensive list of fluorophores and dyes that can be used with the ZE5 Cell Analyzer.
* Small particle detector can be included in configuration or omitted from configuration with the option to add later.
** Eligible for an upgrade with a UV laser.
AFxxx, Alexa Fluor; APC, allophycocyanin; BV, Brilliant Violet; BUV, Brilliant UltraViolet, CFP, cyan fluorescent protein;Cy, cyanine; dsRed, Discosoma Red Fluorescent Protein; FITC, fluorescein isothiocyanate; GFP, green fluorescentprotein; eGFP, enhanced green fluorescent protein; eYFP, enhanced yellow fluorescent protein; LP, long pass;PE, phycoerythrin; PerCP, peridinin chlorophyll protein; RFP, red fluorescent protein; SBB, StarBright Blue; SBV,StarBright Violet; SBUV, StarBright UltraViolet; SBY StarBright Yellow; UV, ultraviolet; YFP, yellow fluorescent protein.

BIO-RAD LABORATORIES, INC. | 7

Visit bio-rad.com/cellanalysis for a wide range of videos, technical reports, published findings, and otherresources describing the Bio-Rad ZE5 Cell Analyzer.
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