WARTHIN STARRY STAT

                                                WARTHIN STARRY STAT

Clean all glassware in potassium dichromate sulphuric acid solution. Wash in tap water and rinse several times in triple distilled water.

Solutions

 1) Acidulated water

                   Triple distilled water          -       1000 CC.
Citric acid (1% or less) quantity sufficient to bring solution to a pH of 3.8-4.4

2) 1% Silver nitrate for impregnation.
3) 2% Silver nitrate solution (developer)
4) Gelatin solution
                                    Sheet gelatin (high grade)               -          10 gm
                                     Acidulated water                              -           200 ml.

5) Hydroquinone
                        0.15% hydroquinone in acidulated water.

Note: - Keep silver nitrate 2%, gelatin 5% and hydroquinone (). 15% in a water                       bath at 54-56°C until developer is made.

Developer
                   Silver nitrate in acidulated water 2%               -        1.5 ml
                    Gelatin in acidulated water 5%                      -        3.15 ml                                      Hydroquinone in acidulated water 0.15%        -        200ml

Combine in the order given, in a container kept in the water bath at 54-56°C. rotating the container after each addition.
Do not prepare in advance.


Procedure

 5-6 control slides should be used at periodic intervals as constant check of . solutions. .

1.    Deparaffinise the section in xylene for 10 mts and bring the section to water
through descending series of alcohol.
2.    Rinse twice in triple distilled water. Handle sections with paraffin coated
forceps.
3.    Place in 1% silver nitrate solution heated in a water bath to 43°C for 30 mts (Prepare developer at this point)
4.     Flood, the sections that have been laid across rods with the developer which must be used as soon as it is mixed. Allow sections to develop until they are light brown or yellow. Check with control.
5.     Rinse quickly and thoroughly in hot tap water (approximately 56°C)
6.    Wash in distilled water.
7.     Dehydrate, clear and mount.


Result

                   Spirochaetes          -        Black
                    Background          -        Pale yellow or brown.



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