A General-Purpose Silver Technique for Peripheral Nerve Fibers in Frozen Sections

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Abstract
For the study of peripheral nerve fibers and endings, frozen sections of material fixed for 1 wk to 2 mo in Richardson''s formalin (prepared from para-formaldehyde and containing 3.4% sucrose) are extracted in saturated boric acid for 1-18 hr at 37[degree]C, or in formolalcohol (4 parts absolute ethanol, 1 part 20% formaldehyde) for 1-2 hr at 37[degree]C. Tissues fixed for 3 mo or more do not require extraction. The sections are placed in 10% AgNo3 for 20-25 min at 37[degree]C, then passed through 4 baths of 10% formaldehyde during 15-30 min. They are developed in a diammino-silver solution prepared from Ag2CO3 or from AgNO3, fixed for 10-30 min in 5% Na2SO3 containing 5% ammonia, washed well, toned for 10 min in 0.5% gold chloride, washed briefly and fixed in 5% Na2S2O3. Finally they are washed, dehydrated, cleared, and mounted in Canada balsam.