Re: Bodians stain

From:=?iso-8859-1?q?Tony=20Henwood?=

Ray,
The following might help:

 LOOT'S MODIFIED BODIAN STAIN FOR NERVOUS TISSUE

NOTES:

The protargol solution used for impregnation is
modified by the addition of hydrogen peroxide to delay
the darkening of the solution and copper nitrate to
improve selectivity.

SOLUTIONS:
          1.   Protargol Solution:
                    Dissolve 0.5g Protargol in 50 ml
distilled water,
                    Add with continuous agitation:
                         1% Copper Nitrate        1
ml. 
                         1% Silver Nitrate        1
ml. 
                         30% Hydrogen Peroxide    0.05
ml.

          2.   Reducing Solution I:
                    Hydroquinone                  0.25
g.
                    Concentrated formalin         2.5
ml.
                    Distilled Water               50
ml.

          3.   1 % aqueous Gold Chloride

          4.   Reducing Solution II:
                    Oxalic Acid                   1 g.
                    Concentrated Formalin         2.5
ml.
                    Distilled Water               50
ml.

          5.   5% Sodium Thiosulphate
STAINING PROCEDURE:

1. Dewax and hydrate sections to distilled water.
2. Place in Protargol Solution  37oC  48 hours.
3. Rinse slides briefly in distilled water.
4. Place in Reducing Solution I  5 minutes.
5. Rinse in water and check microscopically, if
underimpregnated return to Protargol solution for a
further 24 hours. 
6. Tone in Gold Chloride         10 minutes.
7. Rinse in water. 
8. Reduce in Reducing Solution II     10 minutes.
9. Rinse and fix slides in Sodium Thiosulphate  2
minutes.
10. Rinse in water, dehydrate, clear and mount.
RESULTS:
               Neurfibrils, nerve fibers, axons   Dark
red to black
               Collagen                          
Light red or grey.
REFERENCE:
          Loots, J.M., etal (1977) Stain Technol
52(2):85-87.


--- Ray Lopez  wrote: > Help,One of
our Pathologist has just asked for
> bodians stain I have not done
> one in years ,does anyone out there have any
> information/procedures. Any
> help would be greatly appreciated.
>  

=====
Tony Henwood JP BAppSc GradDipSysAnaly CT(ASC)
Laboratory Manager
Histopathology Department
The Childrens Hospital at Westmead
Locked Bag 4001
WESTMEAD, 2145
AUSTRALIA

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