Reference for this nerve stain???

From:Tony Henwood

Reference for this nerve stain???

Hi all,
I have found this technique in the archives of my department and was wondering whether anyone has heard of it. I don;t have any idea of where it originates.

Nerve fibres and axons have the ability to absorb silver ions that are then reduced by hydroquinone.  The fibres are further darkened by oxalic acid.  Selectivity is improved by including gelatine in the silver solution.

Fixation:      10% buffered formalin.
Microtomy:     Paraffin sections at 5um.

1.      Impregnating solution - prepare just prior to use:
        Distilled water                                 50ml
        Silver Nitrate                                  0.1g
        Gelatine                                        0.4g
        Place at 60oC for 15 minutes, mixing twice.

2.      Reducing solution:
        Distilled water                                 50ml
        Sodium Sulphite                                 0.5g
        Hydroquinone                                    0.1g
        Mix, stand for 20 minutes before use.

3.      0.2% Gold Chloride

4.      2% Oxalic acid

5.      2% Sodium Thiosulphate.

1.      Dewax and hydrate sections to distilled water.
2.      Place in impregnating solution, 5 hours - overnight (60oC).
3.      Dip slide in distilled water once.
4.      Place in reducing solution, 2 hours.
5.      Running water, 10 minutes.
6.      Distilled water, 1 minute.
7.      Gold Chloride, 5 minutes.
8.      Distilled water, 1 minute.
9.      2% Oxalic Acid, 2 minutes or longer.
10.     Running water.
11.     2% Sodium Thiosulphate, 5 minutes.
12.     Wash in water, 5 minutes.
13.     Dehydrate, clear and mount.

Axons and nerve fibres          black
Nuclei                          shades of grey


Tony Henwood JP, BappSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager
The Children's Hospital at  Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: (02) 9845 3306
Fax: (02) 9845 3318

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