Flow Cytometer Instrument Setup

Find setup procedures and instructions in order to properly utilize your flow cytometer for reliable data acquisition.

 

Flow Cytometer Instrument Setup

In order to generate reliable data, the flow cytometer must be set up properly before data acquisition. The following sections will address machine setup for the flow cytometers listed below.

List of Flow Cytometers and Possible Configurations

Flow Cytometer

Reporter
Channel

Channel Emission

Classification Channel

Channel
Emission

Compensation
needed?

BD FACSCalibur™
(single laser)

FL2

575 nm

FL3

670 nm

Yes

BD FACSCalibur™ (dual laser)

FL2

575 nm

FL4

660 nm

No*

BD FACSCanto™,
BD FACSCanto™ II

PE

575 nm

APC

660 nm

No*

BD™ LSR, LSRII,
BD LSR Fortessa™

PE

575-585 nm

APC

660 nm

No*

BD FACSAria™

PE

575 nm

APC

660 nm

No*

Cytek® Aurora with YG laser

YG1

575 nm

R4

720 nm

No&

Cytek® Northern Lights and Cytek® Aurora without YG laser

B4

580 nm

R4

720 nm

No&

*Compensation is not required for the specified flow cytometers when set up properly, but is recommended for consistent results.
&Unmixing for Cytek instruments is not required.
For flow cytometers not listed here, the end-user needs to set up the machine following similar guidelines. Please refer to Setup Procedure for Other Flow Cytometers below.
The setup process typically includes the following steps. Please see the detailed setup procedure that follows, regarding your specific instrument.
1) Start up the instrument following the manufacturer’s recommendations.
2) Create a template for data acquisition using your instrument’s data acquisition software. A template is a document or worksheet with density plots that allows the user to perform machine setup and data acquisition.
3) Set up the PMT voltages of each channel to be used for data acquisition using the Setup Beads provided in the kit.
4) Determine whether compensation is needed based on the configuration of your system as shown in the table above. If compensation is needed, perform compensation using the Setup Beads provided in the kit.

Setup Procedure for Attune NxT 
Setup Procedure for FACSCalibur™ with Dual Lasers 
Setup Procedure for FACSCalibur™ with a Single Laser 
Setup Procedure for Beckman Coulter CytoFLEX Flow Cytometer 
Setup Procedure for Beckman Coulter Gallios™ Flow Cytometer 
Setup Procedure for BD FACSAria™, FACSCanto™ and LSR Series 
Setup Procedure for Other Flow Cytometers
Setup Procedure for guava easyCyte™ with a Single Laser
Setup Procedure for guava easyCyte™ with Two Lasers
Setup Procedure for BD Accuri™ C6
Setup Procedure for ACEA NovoCyte™ with Dual Lasers
Setup Procedure for Sony Cell Sorter SH800
Setup Procedure for Sony ID7000 Spectral Cell Analyzer
Setup Procedure for Cytek® Aurora & Northern Lights (2+ Lasers)
Cytek LEGENDplex Experiment Template - No YG Laser
Cytek LEGENDplex Experiment Template - YG Laser
Setup Procedure for Sony SA3800 Spectral Cell Analyzer 
Sony SA3800 LEGENDplex Experiment Template
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