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<DIV><FONT face=Arial size=2>Hi</FONT></DIV>
<DIV><FONT face=Arial size=2></FONT> </DIV>
<DIV><FONT face=Arial size=2>Does anyone have good result with BIELSCHOWSK'S
STAIN ?</FONT></DIV>
<DIV><FONT face=Arial size=2></FONT> </DIV>
<DIV><FONT face=Arial size=2>I am using the technique from Bancroft as follows
on brain tissue of dogs:</FONT></DIV>
<DIV><FONT face=Arial size=2></FONT> </DIV>
<DIV><FONT face=Arial size=2>1. Deparaffinize and hydrate to distilled water.
Wash well in distilled water.</FONT></DIV>
<DIV><FONT face=Arial size=2>2. Place sections in 50 ml of 20% silver nitrate in
the dark at 37 C. Keep this silver solution for </FONT></DIV>
<DIV><FONT face=Arial size=2>stage 4.</FONT></DIV>
<DIV><FONT face=Arial size=2>3. Wash well in distilled water.</FONT></DIV>
<DIV><FONT face=Arial size=2>4. Add strong ammonia to 50 ml of silver nitrate
(from stage 2) until the initial precipitate disappears. </FONT></DIV>
<DIV><FONT face=Arial size=2>Leave section in this solution for 10 minutes at 37
C.</FONT></DIV>
<DIV><FONT face=Arial size=2></FONT> </DIV>
<DIV><FONT face=Arial size=2>I would to understand step 4. Should I add
concentrated ammonia?</FONT></DIV>
<DIV><FONT face=Arial size=2></FONT> </DIV>
<DIV><FONT face=Arial size=2>I added concentrated ammonia 1 to 2 drop at a time.
Immediately I got precipitate that did not disappear.</FONT></DIV>
<DIV><FONT face=Arial size=2></FONT> </DIV>
<DIV><FONT face=Arial size=2>I repeated this step with dilute ammonia getting
the same result. I filtered the soluton and continued the</FONT></DIV>
<DIV><FONT face=Arial size=2>staining and obtained no color on my
axons.</FONT></DIV>
<DIV><FONT face=Arial size=2></FONT> </DIV>
<DIV><FONT face=Arial size=2>I guess I have a problem with step 4. How
much volume of ammonia should be added at what concentration. Is there a
critical point in this staining.</FONT></DIV>
<DIV><FONT face=Arial size=2></FONT> </DIV>
<DIV><FONT face=Arial size=2></FONT> </DIV>
<DIV><FONT face=Arial size=2>Thank you </FONT></DIV>
<DIV><FONT face=Arial size=2></FONT> </DIV>
<DIV><FONT face=Arial size=2></FONT> </DIV>
<DIV><FONT face=Arial size=2>Robin </FONT></DIV>
<DIV><FONT face=Arial size=2></FONT> </DIV>
<DIV><FONT face=Arial size=2><A
href="mailto:Robint@mediom.qc.ca">Robint@mediom.qc.ca</A></FONT></DIV>
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