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Special Stain Solution Protocols Page

Hello,  We have starting adding our solution protocols on a new page.  These are all the protocols that we use now. A professional histology service.

http://histologistics.com/histology/special-stain-solution-protocols/

An Expert Histology Service

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How To Detect Alzheimer’s Disease Using Histology

 Dave Adams and Ben Snyder created the first transgenic mouse model in 1995. Histology played a role in the detection of the amyloid plaques surrounded by dystrophic neurites, and show profound synaptic loss, neurofibrillary tangle formation and gliosis.  The beta amyloid and Tau proteins are detected by antibodies used in immunohistochemistry. There are a whole host of different special stains for neuroglia, astrocytes, myelin sheath, neurofibril tangles and senile plaques.  The Bielschowsy stain shows neurofibrils and senile plaques black while the background is yellow-brown.

http://www.nature.com/nature/journal/v373/n6514/abs/373523a0.html

Beilschowsky Special Stain on Mouse Brain For Neurofibrils and Senile Plaques 20x
Beilschowsky Special Stain on Mouse Brain For Neurofibrils and Senile Plaques 20x

 

 

 

 

 

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Bielschowsky for Neurofibrils and Senile Plaques (Alzheimer)

In addition to the Tau and Amyloid proteins discovered through IHC, this silver stain indicates Alzheimer disease.

Control tissue: Brain Cortex, Spinal Cord

Safety equipment: Work under a hood with lab coat, gloves, and glasses.

  1. De-paraffin slides in xylene (1) —————— 2 minutes (re-use)
  2. De-paraffin slides in xylene (2)——————- 2 minutes (re-use)
  3. Clear slides in 100% alcohol———————- 2 minutes
  4. Clear slides in 100% alcohol———————- 2 minutes
  5. Hydrate slides in 95% alcohol——————– 2 minutes
  6. Hydrate slides in running water —————– 5 minutes
  7. 10% Silver nitrate ——————————  30 minutes in 40C oven
  8. Place slides in DH2O—————————-  until new solution is made
  9. Slowly add ammonium hydroxide to the 10% silver nitrate until it is clear.
  10. Place slides in solution in 40C oven ————– 30 minutes
  11. Place in developer solution ———————- 5 minutes (re-use)
  12. Slides should be golden, use scope to check.
  13. Place in 1% ammonium hydroxide to stop reaction – 1 minute
  14. Wash in water ———————————– 2-5 minutes
  15. Place in 5% sodium thiosulfate ——————- 5 minutes (re-use)
  16. Wash in running water ————————–  5 minutes
  17. Dehydrate in 95% alcohol———————— 30 seconds
  18. Dehydrate in 100% alcohol———————–  1 minute
  19. Dehydrate in 100% alcohol———————–  2 minutes
  20. Dehydrate in 100% alcohol———————–  2 minutes
  21. Clear in xylene (3)——————————-  2 minutes (re-use)
  22. Clear in xylene (4)——————————-  5 minutes (re-use)

Results:           Neurofibrils and senile plaques- black

Background- yellow to brown

 

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Special stains protocols

I am trying to figure out what to do about the special stains protocols.  There are about 35 of them.  Putting them on the front page does not seem to be efficient.   I would like them all to be searchable and downloadable, maybe on a different page.  What do you think, does it make a difference?