A Glutaraldehyde-Dichromate-Silver Sequence for Differentiating Norepinephrine Cells from Epinephrine Cells in Neonatal and Adult Rats—Paraffin Sections |
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Authors: | Philip Ashley Jones |
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Institution: |
a Department of Anatomy, University College, Cardiff, Wales |
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Abstract: | A glutaraldehyde-K2Cr2O7 procedure intensified by silver staining enabled norepinephrine and epinephrine cells to be distinguished readily in paraffin sections of the adrenal glands of rats 8 days after birth. The technique involved fixation in 0.1 M cacodylate-buffered 5% glutaraldehyde (6-24 hr), treatment with 3.5% K2Cr2O7 (6-12 hr) and routine preparation of paraffin sections. The sections were deparaffinised, brought to water and immersed in Fontana's solution (24 hr), prepared by adding concentrated NH4OH drop by drop to 5% AgNO3 until the precipitate formed just redissolved; more 5% AgNO3 was then added until a permanent cloudiness just developed. After a rinse in distilled water, the sections were treated with 0.5% gold chloride (5 min) and Na2S2O3 (5 min), then mounted in Depex. This sequence resulted in an intense black cytoplasmic colouration in norpinephrine-containing cells of both the adult and 8-day-old animals whereas epinephrine-containing cells remained colourless. The glutaraldehyde-K2CrO7 procedure, without intensification, gave very clear results in the adult: a yellow cytoplasmic colour in the norepinephrine cells with epinephrine cells colourless. A glutaraldehyde-OsO4 sequence gave a less well defined separation of these cell types in the adult and failed to distinguish the cell types in the neonate. |
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