The retention of radio isotope-labekd vitamin A during processing for electron microscopy was investigated using the livers and kidneys of vitamin A deficient rats. [15-14C]Retinol (3μCi/animal) was administered by esophageal intubation to male rats which had been maintained on a vitamin A deficient diet for five or sir weeks postweaning. Glutaraldehyde- or osmium-fixed tissue was processed by three methods: a) routine (a graded series of ethanols, propylene oxide and epoxy), b) rapid (75% and 95% ethanol with three changes of epoxy), or c) water-soluble embedding (70% and 80% hydrorypropyl methacrylate). Water-soluble embedding retained the highest percentage of label in the tissue (liver: 96.31%; kidney: 98.68% Inclusion of osmium tetroxide in the processing sequence and minimal exposure of tissue to lipid solvents were necessary for good retention of labeled vitamin A in tissues. The opinions or assertions contained herein are the private views of the author and are not to be construed as official or as reflecting the views of the Department of the Army or the Department of Defense. In conducting the research described in this report, the investigators adhered to the "Guide for Laboratory Animal Facilities and Care," as promulgated by the Committee on the Guide for Laboratory Animal Resources, National Academy of Sciences-National Research Council.
ASJC Scopus subject areas
- Medical Laboratory Technology