Quick sampling and perpendicular cryosectioning of cell monolayers for the X-ray microanalysis of diffusible elements.
A quick sampling and preparation method for freezing of cell monolayers is described. The cells are grown on a large Formvar film supported by a frame of polystyrene. A polyvinylpyrrolidone (PVP) solution is applied to one side of the film forming a flat disc when frozen with a pair of pliers precooled in liquid nitrogen. The PVP solution provides the specimen with sufficient strength and may be used as an elemental standard for absolute quantification if salts of known concentrations are added. Manipulation of the cells prior to freezing is thus restricted to a minimum, which eliminates possible harmful treatments like scraping and centrifugation. The procedure is quickly performed, the freezing being completed within 30 s of the cells having been removed from the culture well. The analytical results reveal low and stable Na:K ratios. Our results confirm that cells in vitro are comparable to cells in vivo with respect to elemental composition.
Tvedt
K E
KE
Department of Pathology, University of Trondheim, Norway.
Halgunset
J
J
Kopstad
G
G
Haugen
O A
OA
D000230:Adenocarcinoma; D004577:Electron Probe Microanalysis / Q000379:methods*; D005615:Freezing; D005629:Frozen Sections; D006801:Humans; D007951:Leukemia, Myeloid; D008297:Male; D008867:Microtomy; D009994:Osmolar Concentration; D011188:Potassium / Q000032:analysis*; D011205:Povidone; D011471:Prostatic Neoplasms; D012964:Sodium / Q000032:analysis*; D014407:Tumor Cells, Cultured
D012964:Sodium; D011205:Povidone; D011188:Potassium