When to use Flow Cytometry?

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When to use Flow Cytometry

What is immunophenotyping?

The urge to detect the elevated number of cell subsets in the immune system is the reason for multicolour flow cytometry to become the most common practice in identifying cell markers.



Immunophenotyping can be a simple cell identification by only a single marker or a complex procedure, involving homing profile, cytokine release and activation sites in a single panel. Apart from research, this technique is also quite useful in clinic diagnostics or routine monitoring. 

What is apoptosis? 

This is the process of programmed cell death, playing an important part in embryogenesis and removing damaged cells. Many cancers have been linked to irregularities in the high regulation of this process, same as autoimmune diseases. 

How is apoptosis measured?

The most common feature of this process can be measured using flow cytometry. This feature is the externalization of phosphatidylserine, commonly found in the inner membrane of healthy cells. By using different dyes and markers, healthy cells can be distinguished from apoptotic and necrotic ones. 

How can cell proliferation be measured?

This process can be measured with flow cytometry. Apart from using the differences in forward and side scatters to look for small changes, using a proliferation marker to stain the cells or DNA markers has far better results. 

Using cytoplasmic dyes

Another way to measure proliferation is by using cytoplasmic dyes. After incubation with these protein-binding dyes, cell division becomes more visible. This is because when the cell divides, the concentration in the new cells is halved, allowing us to observe the reduced fluorescence levels in the following generations. 

How to determine in which stage is the cell?

By using DNA binding dyes, we can determine the stage. For example, cells in G2 have twice the amount of DNA compared to G1, leading to them being two times brighter. Newer software also has the option to estimate the phases by using algorithms.