Staining Paraffin Sections with Protargol 3. The Optimum pH for Reduction. 4. A Two-Hour Staining Method

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Abstract
Further work on conditions affecting the reduction of paraffin sections impregnated with protargol showed that the opt. pH for sulfite-amidol mixtures was between 6.5 and 7.5. A staining method which requires about 2 hours to complete consists of the following steps: (1) One hour impregnation at 60[degree] C in 10% AgNO3. (2) Wash in distilled water 3 changes of 30 sec. each. (3) Put into protargol (Winthrop Chem. Co., New York, N. Y.) 0.2% aq. for another hour at room temp. (4) Rinse 2 sec. (5) Reduce 1-2 min. in amidol 0.2 g., Na2SO3, 8 g., NaHSO3 1 g., and water 100 cc. (6) Wash thoroughly. (7) Tone with 0.1% gold chloride. (8) Wash. (9) Reduce with a 0.5% aq. soln. of amidol (no sulfite). (10) Wash, dehydrate and cover. The method stains neurofibrillae and unmyelinated fibers . and has worked well on most tissues of vertebrates. The stain follows acid alcoholic fixation.