| Von
Kossa’s Staining Procedure for Calcium
Literature
Purpose:
This standard operating procedure (SOP) establishes the standard
for staining calcium using the Von Kossa’s staining procedure.
Reagents
and Materials
Reagents
5%
Silver Nitrate
Silver Nitrate
5.0gm
Distilled water 100ml
2% Sodium Thiosulfate
Sodium Thiosulfate
2.0gm
Distilled water 100ml
Nuclear
Fast Red
Commercially
prepared by Newcomer Supply Co. (catalog # 12552)
Expiration date provided by manufacturer.
Store at room temperature.
Materials
Tissue slides and control slides for special stain requested.
Microwave to bake slides.
Staining rack, 20 slide capacity.
Coverslips and synthetic mounting media.
Hazards
and Precautions:
See Material Safety Data Sheets.
Sample Preparations:
Paraffin tissue section slides cut at 4 microns.
Calibration
and Standardization:
A tissue section slide containing calcium is stained with each procedure.
Procedure:
1. Deparaffinize and hydrate to distilled water.
2. Place in 5% Silver Nitrate in a plastic coplin jar with lid applied
loosely.
Microwave at 70% power for 1 minute.
3. Distilled water rinse.
4. 2% Sodium Thiosulfate – 2 minutes.
5. Distilled water rinse.
6. Nuclear Fast Red – 5 minutes.
7. Distilled water rinse.
8. Dehydrate, clear and coverslip.
 Positive
control: Canine calcinosis circumscripta (Figure
1)
Results:
Calcium salts – dark brown
Background – varying shades if pink
Cherie
Chapman
James R. Turk
7/1/04
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