"Liquidless" cell staining by dye diffusion from gels and analysis by laser scanning cytometry: potential application at microgravity conditions in space.

摘要: Background Conventional staining of cells or tissue sections on microscope slides involves immersing the into solutions dyes then rinsing to remove unbound dye. There are instances, however, when use stain is undesirable—e.g., at microgravity conditions in space, where possibility accidental spill (many known carcinogens) introduces health hazard. Likewise, transporting bulk liquid stains and rinses may be burdensome certain situations such as field expeditions combat. Methods The “liquidless” procedure proposed which contained thin strips hydrated polyacrylamide gelatin gels that have been presoaked solutions. Fluorochromes affinity DNA (propidium iodide, PI; 4,6-diamidino-2-phenylindole, DAPI, Hoechst 33342) protein (sulforhodamine 101) were used saturate gels. The gel placed over prefixed deposited relatively low (20 g/cm2) pressure was applied ensure contact. also stained by using commercially available mounting media DAPI PI admixed. Intensity fluorescence measured laser scanning cytometry (LSC). Results Satisfactory cell staining, with minimal background, achieved after 10–20 min contact between Optimal concentrations presoak found 2–4-fold higher than routinely cytometry. measurements intensity cellular LSC revealed stoichiometric reflected characteristic content frequency histograms distinct G1, S, G2/M populations 2:1 ratio G1 peak fluorescence. Individual can saturated more a single dye—e.g., obtain differential staining. Cell gelatin-based led high background while “aqueous” medium satisfactory. Conclusions Relatively fast nonconvective dye diffusion permeated dyes, pressure. quality provided this methodology comparable conventional Cytometry 44:355–360, 2001. © 2001 Wiley-Liss, Inc.