Re: [Histonet] Bielschowsky's stain


I have a very nice Bielchowsky's method I used at University of Chicago. It
always worked.  If you like I can email it to you.


Mari Ann Mailhiot BA HT ASCP
Application Specialist
Leica Technical Assistance Center
800 248 0123 x7267
847 236 3063 fax

                      "Robin Turcotte"                                                                                                        =20
                                     To:                                                   =20
                      Sent by:                            cc:                                                                                 =20
                      histonet-admin@lists.utsouth        Subject:  [Histonet] Bielschowsky's stain                                           =20
                      11/19/2003 06:54 PM                                                                                                     =20


Does anyone have good result with BIELSCHOWSK'S  STAIN ?

I am using the technique from Bancroft as follows  on brain tissue of dogs:

1. Deparaffinize and hydrate to distilled water.  Wash well in distilled
2. Place sections in 50 ml of 20% silver nitrate in  the dark at 37 C. Keep
this silver solution for
stage 4.
3. Wash well in distilled water.
4. Add strong ammonia to 50 ml of silver nitrate  (from stage 2) until the
initial precipitate disappears.
Leave section in this solution for 10 minutes at 37  C.

I would to understand step 4. Should I add  concentrated ammonia?

I added concentrated ammonia 1 to 2 drop at a time.  Immediately I got
precipitate that did not disappear.

I repeated this step with dilute ammonia getting  the same result. I
filtered the soluton and continued the
staining and obtained no color on my  axons.

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

Thank you


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