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This flow cytometry guide aims to give you a basic overview of all the important facets of flow cytometry without delving too deeply into the complex mathematics and physics behind it all. Instead, we present a guide that will be invaluable to beginners of flow cytometry or act as a fact-packed synopsis for those of you interested in teaching others about the virtues of this powerful application.
This chapter explains how a flow cytometer works. You will learn how the cells pass through the instrument, how light is detected and measured and the basic principles behind sorting cells.
Fluorophores are fluorescent markers which absorb light energy and emit at a longer wavelength. This chapter explains how they work, why fluorescent markers are so important in flow cytometry and how to compensate between them.
Flow cytometry data can be analysed in different ways. Here we give common examples of gating strategies and how data can be best represented.
The quality of data you obtain from flow cytometry can be affected by both the health of the cells and the staining protocol used. Here we give examples of commonly used cell preparation techniques and staining protocols for both surface and intracellular markers.
There can be many reasons for poor results. Here we list some of the common problems with advice to help you get the best staining possible along with recommended reading if you want more information.
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