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Testimonials

inverted CommasI can’t recommend FloCyte enough to people. I wish everyone who prepares flow samples – even if they don’t run them – would take FloCyte’s courses

 

 

Compensation and Multiparameter Strategies

Our first course in Multiparameter Flow Cytometry back in 2004, taught by Dr. Carleton Stewartwho passed on in 2015, was the Director of the Flow Cytometry Lab of Roswell Park Cancer Center in Buffalo, New York.  This was a great success.  We have continued to build this curriculum begun with Dr. Stewart’s presentations on the basics of antibody binding kinetics, titering antibodies, extra and intracellular staining, using tandem dyes, blocking,  and viability and have augmented it with a complete compensation presentation, built on the work of Mario Roederer, Wayne Moore and Dave Parks,and Dr. Bartek Rajwa’s work on spectral unmixing. The response to this course has been excellent!

 

Our Full day Curriculum covers these topics:

 

Introduction to the course:
Physics of light

Fluorescence

Fluorochromes and Reagents

Considerations for selecting reagents
Viability

Controls

Beads for Instruments and assays
Isotypes vs No isotypes
FMOs
Compensation Controls

Staining Methodology

Titering and Saturation Issues
Autofluorescence Problems
Specific VS Non- specific fluorescence

Compensation

In Multicolor phenotyping
How to do it correctly
Measurement Error
Hardware vs Software
Bi-Exponential Scaling
Compensation Controls
Spectral Unmixing vs Compensation

Strategies for Building Multicolor Panels

Antibody / Fluorochrome combinations
Necessary, Important, or Luxury Markers
Calculating Spread and Error
Instrument Configurations / Filter Choices

Practice Designing Experiments

Design Process
Biological hypothesis

We can also present a Half Day Program, concentrating on the strategies for building multicolor panels And issues involved in designing them!