The technique you describe (quoted below, with a
reference to Garvey, Bigelow & Carpenter 1999) closely 
resembles an earlier one published by Winsome Garvey 
et al 1987, in J. Histotechnol. 10:245-247. It bears 
no resemblance whatever to any of the techniques
introduced by Bielschowsky.

The "parent" method would probably be either that
of EP Samuel 1953 J. Anat. 87:278-287 or that of
A Peters 1955 Quart. J. Micr. Sci. 96:323-328. 
These are "physical developer" techniques for
staining axons. (A physical developer is one
that contains silver ions and a reducing agent
together with a stabilizing colloid - in this
case, gum mastic - to slow down the otherwise
almost instantaneous reaction between them.)

The distinguishing feature of Garvey's method is 
the use of a high temperature to allow a short 
time and low silver concentration in the first 
stage of the procedure. Garvey et al's paper in
J Histotechnol 22:37-41 (1999) provides many
additional technical tips as well as some good 
discussion of how and why the method works, 
and 31 references.
-- 
-------------------------
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1
   kiernan@uwo.ca
   http://publish.uwo.ca/~jkiernan/
________________________________________________
"Featherstone, Annette" wrote:
> 
> Here is the Alternate Bielschowsky Stain:
> 
> 1. Deparaffinize to DH2O
> 
> 2. Place sections in 0.1% silver nitrate solution, containing citric acid
> (working silver nitrate) in a 65 degree C water bath for seven minutes. (I
> preheat this solution for about 5min, in a plastic coplin jar)
> 
> 3. Rinse in 2 changes of DH2O, remove slides one at a time.
> 
> 4. Rinse in 95% ethanol, then 2 changes of AbsoluteEthanol.
> 
> 5. Immerse slides in filtered gum mastic for about 5 min. (We keep this in a
> coplin jar and top it off as needed)
> 
> 6. Drain well and put into developing solution in a 65 degree C water bath
> for 12- 13 minutes. (I start to check it at around 10 minutes, I also
> preheat this solution for about 5 min, usually when I put the slides in the
> gum mastic.
> 
> 7.Rinse in DH2O
> 
> 8. Dehydrate clear and mount.
> 
> SOLUTIONS:
> 
> 1% Silver Nitrate
>       Dissolve 1gm silver nitrate in 100 ml DH20. this solution last for
> months in a brown bottle in the frig provided it is not left
>       at room temp.
> 
> 0.1% Silver Nitate
>       Take 5ml of the 1%silver nitrate, dilute with 45 ml DH2O. Prepare
> fresh each time
> 
> 1% Citric Acid
>       1gm citric acid in 100 ml DH2O
> 
> Working Silver Nitrate Solution
>       Add 0.1 ml of 1% citric acid to 40 ml of the 0.1% silver nitrate.
> 
> Gum Mastic
>      Dissolve 2.5 gm of gum mastic in 100ml absolute ethanol. Filter before
> use. May be topped off and reused.
> 
> Hydroquinone
>     Dissolve 0.5gm Hydroquinone in 30 ml DH2O
> 
> Developing Solution
>     Combine 10 ml filtered gum mastic, 30 ml Hydroquinone, add 0.9ml of 1%
> silver nitrate
> 
> We got this stain from the Journal of Histotechnology/Vol. 22, No. 1/ March
> 1999 and we have used it ever since with absolutely beautiful results
> 
> We always use sterile water but I guess DH2O might be fine too. This saves
> hundreds of dollars per year, maybe more.
>  Here is the Alternate Bielschowsky Stain:
> 
> 1. Deparaffinize to DH2O
> 
> 2. Place sections in 0.1% silver nitrate solution, containing citric acid
> (working silver nitrate) in a 65 degree C water bath for seven minutes. (I
> preheat this solution for about 5min, in a plastic coplin jar)
> 
> 3. Rinse in 2 changes of DH2O, remove slides one at a time.
> 
> 4. Rinse in 95% ethanol, then 2 changes of AbsoluteEthanol.
> 
> 5. Immerse slides in filtered gum mastic for about 5 min. (We keep this in a
> coplin jar and top it off as needed)
> 
> 6. Drain well and put into developing solution in a 65 degree C water bath
> for 12- 13 minutes. (I start to check it at around 10 minutes, I also
> preheat this solution for about 5 min, usually when I put the slides in the
> gum mastic.
> 
> 7.Rinse in DH2O
> 
> 8. Dehydrate clear and mount.
> 
> SOLUTIONS:
> 
> 1% Silver Nitrate
>       Dissolve 1gm silver nitrate in 100 ml DH20. this solution last for
> months in a brown bottle in the frig provided it is not left
>       at room temp.
> 
> 0.1% Silver Nitate
>       Take 5ml of the 1%silver nitrate, dilute with 45 ml DH2O. Prepare
> fresh each time
> 
> 1% Citric Acid
>       1gm citric acid in 100 ml DH2O
> 
> Working Silver Nitrate Solution
>       Add 0.1 ml of 1% citric acid to 40 ml of the 0.1% silver nitrate.
> 
> Gum Mastic
>      Dissolve 2.5 gm of gum mastic in 100ml absolute ethanol. Filter before
> use. May be topped off and reused.
> 
> Hydroquinone
>     Dissolve 0.5gm Hydroquinone in 30 ml DH2O
> 
> Developing Solution
>     Combine 10 ml filtered gum mastic, 30 ml Hydroquinone, add 0.9ml of 1%
> silver nitrate
> 
> We got this stain from the Journal of Histotechnology/Vol. 22, No. 1/ March
> 1999 and we have used it ever since with absolutely beautiful results
> 
> We always use sterile water but I guess DH2O might be fine too. This saves
> hundreds of dollars per year, maybe more.
> 
> 
> -----Original Message-----
>>> From: J. A. Kiernan [mailto:jkiernan@uwo.ca]
>>> Sent: Friday, February 14, 2003 14:24
>>> To: Nymeyer, Heather
>>> Cc: 'histonet@pathology.swmed.edu'
>>> Subject: Re: Bielchowsky stain
>> 
> > "Nymeyer, Heather" wrote:
> >  ... The method being used is a 20 % silver nitrate
>>> > solution for 20 mins followed by a 15 min incubation
>>> > (in the dark) of ammoniacal silver. ... A few drops of the reducer
>>> > is added to the ammoniacal silver and checked microscopically
>>> > until fibers are seen.  At the same time the fibers are being
>>> > developed the excess background precipitate is forming.
> >
>>> This is what to expect if you add formaldehyde to an
>>> ammoniacal silver solution; it's one way of making
>>> mirrors. In all the variants I've encountered (and most
>>> bear little resemblance to the methods published by
>>> Max Bielschowsky from 1904-1910), the sections are put
>>> in the reducing solution after the ammoniacal silver.
>>> There may or mmay not be a distilled water rinse
>>> between these steps, and this is a critical feature
>>> affecting the result. A small amount of ammonia-silver
>>> complex must be carried over, either as free liquid or
>>> not washed out of the tissue.
>> 
> The principle of the method is that silver precipitation
> occurs fastest in the axons because these already
> contain small amounts of colloidal silver, formed
> during the immersion in silver nitrate. Eventually
> there will be silver precipitation everywhere, hence
> the need for microscopic control of the reduction
> step. The rate of reduction is affected by a variety
> of factors such as the concentrations of silver and
> ammonia and the pH. (These are explained by Nauta WJH
> & Gygax PA (1951) Stain Technol. 26:5-11 in a paper
> about their Bielschowsky-like method for degenerating
> axons.)
> 
> If you're staining normal axons there are easier
> silver methods than those based on Bielschowsky's.
> 
> --
> -------------------------
> John A. Kiernan
> London,   Canada   N6A 5C1
__________________________________________________