Appendix B Example 9-Color Immunophenotyping Experiment; Preparing Controls And Samples - BIO RAD ZE5 User Manual

Appendix B
Experiment
This section provides an example of how to set up a typical multicolor experiment using the ZE5™
Cell Analyzer and Everest™ Software. It shows how to set up compensation controls using the
compensation template and how to apply auto compensation. The sample experiment uses the
following fluorophores:
n
Alexa Fluor 488
n
Brilliant Ultraviolet (BUV) 395
n
APC-Cy7
n
PE (R-phycoerythrin)
n PE-Cy7
n
PE-Dazzle 594
n Brilliant Violet (BV) 421
n
Brilliant Violet (BV) 510
n
Brilliant Violet (BV) 711

Preparing Controls and Samples

First, prepare the compensation controls and samples to be placed in your medium of choice, such as
a 40-tube rack.
1. Prepare a compensation control for each fluorophore to be used in the experiment.
2. Prepare an unstained cell sample.
3. Prepare a fully stained test sample.
Example 9-Color Immunophenotyping
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