Bielschowsky's silver stain
Paul Polak and Douglas Feinstein, University of Illinois, Chicago. Bielschowsky's silver stain is a very useful tool to detect nerve fibers. It can be used to stain axons, neurofibrils, and senile plaques, which are commonly associated with neurodegenerative conditions such as Alzheimer's disease.
Last edited Mon 29 Aug 2022
Stage 1 - Bielschowsky’s silver stain
Steps
Deparaffinize and hydrate sections to dH2O. Wash 3X for 3 min in dH2O.
Place sections in 50 mL 10% silver nitrate in the dark at 37ºC for 30 min.
- Keep this solution after incubation (for use in step 4).
Wash 3 times for 3 min in dH2O.
Add concentrated ammonium hydroxide dropwise with stirring to the silver nitrate solution reserved from step 2.
- Add only enough to dissolve the dark initial precipitate but not more.
Incubate sections in this solution for 15 min at 37ºC.
- Again, save this solution for use in step 7.
Wash sections in 0.1% ammonium hydroxide 3 times for 2 min at room temperature.
Add 350 μL developer solution (0.2 mL 37% formaldehyde, 12 mL dH2O, 12.5 μL 20% nitric acid, and 0.05 g citric acid) to the silver hydroxide solution saved from step 4.
Stain sections in this solution for 10 min until they turn black.
Wash in 0.1% ammonium hydroxide 3 times for 2 min and dH2O 3 times for 2 min.
Tone in 0.2% gold chloride for 5 min.
Fix in 5% sodium thiosulfate for 1 min.
Wash in dH2O, dehydrate in alcohols, then xylene, and mount.
- Figure 1. Example of immunohistochemistry using the Bielschowsky Silver Stain Kit, staining formalin-fixed, paraffin-embedded human brain.
