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sashiolina
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 Inserito il - 04 settembre 2009 : 20:41:54
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cerca i protocolli per il Von Kossa
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Description: This technique is for demonstrating deposits of calcium or calcium salt so it is not specific for the calcium ion itself. In this method, tissue sections are treated with a silver nitrate solution and the silver is deposited by replacing the calcium reduced by the strong light, and thereby visualized as metallic silver.
Fixation: formalin fixed, paraffin embedded tissue sections or alcohol fixed, frozen sections.
Solutions and Reagents:
1% Aqueous Silver Nitrate Solution:
Silver nitrate ------------------------- 1 g
Distilled water ---------------------- 100 ml
5% Sodium Thiosulfate:
Sodium thiosulfate ---------------- 5 g
Distilled water -------------------- 100 ml
0.1% Nuclear Fast Red Solution:
Nuclear fast red ------------------- 0.1 g
Aluminum sulfate------------------ 5 g
Distilled water ----------------------100 ml
Dissolve aluminum sulfate in water. Add nuclear fast red and slowly heat to boil and cool. Filter and add a grain of thymol as a preservative.
Procedure:
1.
Deparaffinize paraffin sections and hydrate to water.
2.
Rinse in several changes of distilled water.
3.
Incubate sections with 1% silver nitrate solution in a clear glass coplin jar placed under ultraviolet light for 20 minutes (or in front of a 60-100 watt light bulb for 1 hour or longer).
4.
Rinse in several changes of distilled water.
5.
Remove un-reacted silver with 5% sodium thiosulfate for 5 minutes.
6.
Rinse in distilled water.
7.
Counterstain with nuclear fast red for 5 minutes.
8.
Rinse in distilled water.
9.
Dehydrate through graded alcohol and clear in xylene.
10.
Coverslip using permanent mounting medium.
Results:
Calcium salts ------------------------ black or brown-black
Nuclei -------------------------------- red
Cytoplasm --------------------------- pink |
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protocollo colorazione
Didattica e Relazioni
