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A 3-step procedure performed at 85 C was as follows: Sections were treated with 4% aqueous ferric ammonium sulfate for 15-300 sec, washed, placed in ripened, 0.5% hematoxylin for 15-300 sec, washed, and counterstained in 1% Safranin O for 15-90 sec. This procedure produced a durable stain with clear cytoplasmic and nuclear differentiation.
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