⑫ Staining

Kossa silver impregnation method

Calcium salt is stained a deep brown black color.

Staining procedure

1DeparaffinizationXylene3 changes, 10 minuntes each
2Removal of xylene100% ethanol3 changes, 5 minutes each
3Hydration95%, 70% ethanol5 minutes each
4WashingRunning tap water 2 minutes
5RinsingDistilled water 
6Silver impregnation5% silver nitrate solutionUnder indirect light for 10 minutes to 1 hour
7WashingDistilled waterSeveral changes
8Fixing5% sodium thiosulfate solution1 minute
9WashingRunning tap water10 minutes
10CounterstainingKernechtrot5 minutes
11WashingRunning tap water10 minutes
12Dehydration70% , 95% ethanol5 minutes each
13Dehydration100% ehtanol3 changes, 5 minutes each
14ClearingXylene3 changes, 10 minuntes each

Staining method

  1. Deparaffinization
  2. Wash in distilled water
  3. Incubate in 5% silver nitrate under indirect light for 10 minutes to 1 hour
  4. Place in 5% sodium thiosulfate for 1 minute
  5. Flushing
  6. Nuclei are stained red with Kernechtrot.
  7. Flushing
  8. Dehydration, penetration, and encapsulation

Staining results

Areas of calcification are stained black.

Calcium phosphate is mainly targeted.

Please be aware that copper, mercury, and lead also react with this method.