Flow Cytometry

How works flow cytometry?

How to apply two different lazers (e.g., FITC and PE) at the same time to excite cells, and then detect and sort only cells that are excited by both lazers, and get data from computer?

GFP is the easiest excitation since it is much less toxic than PE excitation that might kill your cells.

This would be the same effect by Image J. (e.g. Getting FITC and PE images, respectively from fluorescence microscope and then merge them, which generates yellow pseudo color)

To know more about BD Facs follow the advice of Prof. Dr. Annelise Snyder from th UCSD Extension School in her course of Applied Immunology (BIOL-40371) recorded in Summer Quarter 2021.
This lecture provides an introduction to the BD flow cytometry techniques used for immunophenotyping both in basic research and clinical patient samples.
This lecture includes a discussion of sample Beckton Dickinson flow plots used to diagnose various clinical pathologies, including B cell acute lymphoblastic leukemia (B cell ALL), severe combined immunodeficiency (SCID), perforin deficiency, and chronic granulomatous disease (CGD).
Spherotech callibration with AccuCount and Rainbow callibration.
All figures are either from Janeway's Immunobiology (9th ed.) where noted, or my own original figures.