Staining Residual Lipids in Ultrathin Sections of Tissues Embedded in Polyester Resin |
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Authors: | Helene Tzitsikas E. J. Rdzok A. E. Vatter |
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Affiliation: | a Research Division, Abbott Laboratories, North Chicago, Illinois |
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Abstract: | Rat suprarenal glands fixed in Palade's 1% OsO4, buffered at pH 7.7 with veronal-acetate, to which 0.1% MgCl2 was added, were embedded in Vestopal-W and sectioned at 0.2-1 µ. The sections were attached to slides by floating on water, without adhesive, and drying at 60-80° C, placed in acetone for 1 min and then treated with the following staining procedure: Place the preparation in a filtered solution of oil red O, 1 gm; 70% alcohol, 50 ml; and acetone, C.P., 50 ml; for 0.5-1 hr. Rinse in absolute ethyl alcohol; drain; counterstain with 0.5% aqueous thionin for 5 min; rinse in distilled water; drain; stain in 0.2% azure B in phosphate buffer at pH 9, for 5 min. Dry and apply a drop of immersion oil directly on the section. The preparations are temporary. Ciaccio-positive lipids, rendered insoluble by OsO, fixation, stained red to ochre. |
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