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Presented by: Dr Mike Blundell, Product Manager at Bio-Rad
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 in flow cytometry and act as a fact-packed synopsis for those of you interested in teaching others about the virtues of this powerful application.
To further assist learning we have recently added a Flow Cytometry glossary of terms to this guide.
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 analyzed in different ways. Here we give common examples of gating strategies and how data can be best represented.
Controls are vital in any experiment to reliably distinguish your results from background variation and non-specific effects. Here we will discuss some essential controls for flow cytometry you must consider to ensure publication quality flow cytometry data.
One of the fundamentals of flow cytometry is the ability to measure single particles as they pass through the laser, however, if you start with a poor sample it is likely that the data collected will be poor as well. In this chapter we give you advice on how to prepare and treat samples to ensure you have a viable cell suspension.
Building complex multicolor panels can be difficult. This chapter gives some basic rules you should follow in your experimental design to build multicolor panels.
Flow cytometry has many varied uses. In this section we will briefly discuss popular techniques currently being used in flow cytometry to distinguish one particle from another and some of the emerging technologies.
The data of quality from flow cytometry can be affected by both your cell health and staining protocol used. Here we give examples of commonly used cell preparation techniques and staining protocols for surface markers, intracellular markers and DNA.
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.