Intracellular fluorescent labelling of cells for analysis of lymphocyte migration.
A procedure for analysing in vivo migration of lymphocytes labelled in vitro using intracellular fluorochromes is described. Comparison of carboxyfluorescein diacetate, a cytoplasmic label, with Hoechst dye No. 33342 (H33342), a DNA-binding fluorochrome, indicated that H33342 is superior. The concentration of H33342 used for labelling does not significantly affect viability or lymphocyte migration and permits long-term visualization. H33342 allows quantitation of in vivo migration in cell suspension and histological localization in frozen sections. Fluorescence is retained in fixed frozen sections for at least 3 months. This method can be used for analyses of lymphocyte migration and maturation.
D000230:Adenocarcinoma / Q000276:immunology; D000818:Animals; D001562:Benzimidazoles; D002460:Cell Line; D002465:Cell Movement; D005260:Female; D005452:Fluoresceins; D005456:Fluorescent Dyes; D007700:Kinetics; D008214:Lymphocytes / Q000166:cytology* / Q000276:immunology; D008297:Male; D008325:Mammary Neoplasms, Experimental / Q000276:immunology; D012515:Mast-Cell Sarcoma / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence; D051381:Rats; D013154:Spleen / Q000166:cytology / Q000276:immunology
D001562:Benzimidazoles; D005452:Fluoresceins; D005456:Fluorescent Dyes; C027780:carboxyfluoresceindiacetate; C017807:bisbenzimide ethoxide trihydrochloride