Flow cytometry (FCM) of human epidermal cells. A preparation method for epidermal cells and demonstration of circadian variations in the proportion of S-phase cells.
A suction blister technique on the abdominal skin to separate pure human epidermis from its basement membrane is described. After incubation with proteolytic enzymes (trypsin, pepsin) and mechanical shaking, isolated cells and nuclei were stained with ethidium bromide. DNA-specific fluorescence was measured in a flow cytometer using UV excitation light. Cell-cycle phase distributions were obtained from the resulting histograms using a planimetric method. The proportion of cells with an S-phase DNA content showed circadian variations with peak values shortly before noon, and lower values during the rest of the 24 h period.
D001768:Blister / Q000473:pathology; D002455:Cell Division; D002467:Cell Nucleus / Q000378:metabolism; D002469:Cell Separation / Q000379:methods*; D002940:Circadian Rhythm; D000078404:Epidermal Cells; D006801:Humans; D007399:Interphase; D013936:Thymidine / Q000378:metabolism
D013936:Thymidine