https://onlinelibrary.wiley.com/doi/full/10.1002/eji.201646632#

https://www.thermofisher.com/es/es/home/life-science/cell-analysis/cell-analysis-learning-center/molecular-probes-school-of-fluorescence/flow-cytometry-basics.html

Cytometry —> measurement of cell characteristics (size, cell count, cell cycle, etc)

Makes the cells line up one after the other with laminal flow.

Laser trough the cells and some structures make it scatter.

SCATTER PLOTS

• FOWARD SCATTER: size of the cell. Larger cells have a larger foward scatter. FSC (x)
• SIDE SCATTER: granularity. Cells with more stuff inside have larger side scatters. SSC (y)

(blood scatter - lymphocites with low and side scatter, monocites slightly more increased with the side scatter and foward, granulocytes wich have a large side scatter)

Detecting Flourescence

Antibodies specific to label cellular antigents

As the flourence is imperfect and has values that overlap you do compensation:

• calculate the overall spillage of the FITC signal into the PE channel

Take the % and substract it from the channel that is being overlaped.

Plotting and gating

Lynphocites are marked with CD3.

3 dimensions