Cell cycle analysis with flow cytometry & propidium iodide ... | 合法醫療器材資訊網

Introductiontoflowcytometricanalysis...CellcycleanalysisbyquantitationofDNAcontentwasoneoftheearliestapplicationsofflowcytometry.TheDNAof ...

DNA-binding dyes include propidium iodide (PI)[1], 7-aminoactinomycin-D (7-AAD)[2], Hoechst 33342[3], 33258[4] and S769121[5], TO-PRO-3, 4’6’-diamidino-2-phenylindole (DAPI)[6], DRAQ5™[7] and DRAQ7™[8].

In most cases, cells must be fixed or permeabilized to allow entry of the dye which is otherwise actively pumped out by living cells.  For fixation, alcohol or aldehyde are commonly used. Alcohol is a dehydrating fixative which also permeabilizes. This will allow easy access of the dye to the DNA and gives good profiles (low coefficient of variation, CV). The disadvantage is that it is often incompatible with fluorescent proteins and some surface markers.

If fluorescent proteins or surface markers need to be examined simultaneously, use of an aldehyde (cross-linking) fixative, usually paraformaldehyde is more appropriate. This may lead to poorer quality profi...