SAMPLE DEGRADATION BY FLOWCYTOMETRY:

October 6, 2015

 

Theoretically  any type of sample can be used for flow cytometry as all that is required is a single cell suspension of intact cellular structures. We routinely run various immune preparations, and they can be performed both fresh  or  fixed (when performing immunophenotyping). The sample frozen in  any cryopreservation agent such as DMSO are suitable since the cells are intact otherwise sample alone frozen is likely to be as “dead”. The Frozen sample would first have to be thawed before running them on flow, but as long as the cells are intact there should not be a problem, for performing immunophenotyping, so you could thaw the cells, stain them with respective antibodies, and then fix them before analysis.  The RBC's in whole blood can sometimes interfere with analysis because they are in such high abundance, so perform a RBC lysis stage (or a Ficoll gradient) to remove these and analyze the remaining white blood cells only. For granulocytes, you have to use fresh blood, since they don't survive freezing. An RBC lysis step is also required to get good results.

As in our lab we are using mostly fresh samples but some experiment should perform to show the sustainability bone marrow or peripheral blood that for how long we can use the sample at 4-5Oc temperature so to saw the result we have done an experiment  discussed below

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