Preservation of cells sorted individually onto microscope slides with a fluorescence-activated cell sorter.
Fluorescence-activated cell sorters permit analyses and separation of cell populations based on light scatter and surface immunofluorescence parameters. Since a sorter can deposit individually identifiable cells onto a microscope slide, it was considered of interest to combine the flow measurements with analyses available on cells adhering to a surface as in, for example, morphological studies, cytoplasmic immunofluorescent staining, and mRNA in situ hybridization. A necessary condition for these studies is the preservation of cell structures after sorting. We report here a procedure suitable for this purpose. The most important features of this procedure are A) reducing the saline content of the sorter sheath fluid to about 0.0015 M (one-hundredth that of normal saline) to prevent cell damage due to hypertonicity during drying, and B) coating the substrate with a thin layer of newborn calf serum to promote the adherence of the cells to the substrate during subsequent fixing and staining.
D000818:Animals; D002469:Cell Separation / Q000379:methods; D005260:Female; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods; D008214:Lymphocytes / Q000166:cytology*; D051379:Mice; D008815:Mice, Inbred Strains; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000235:genetics; D013154:Spleen / Q000166:cytology; D014021:Tissue Preservation / Q000379:methods*
D012333:RNA, Messenger