Histology

scientificprotocols authored about 3 years ago

Procedure:

  1. Incubate sections in the incubating medium at room temperature for 1 - 3 hours. Two hours is sufficient in most cases.
  2. Wash in distilled water for 2 minutes.
  3. Counterstain with Nuclear Fast Red for 5 - 10 minutes.
  4. Wash in distilled water for 2 minutes.
  5. Air dry and coverslip.

Results:

  1. Nuclei - red
  2. Sites of enzyme activity - blue
  3. To preserve the reaction product, selection of the right mounting (coverslipping) medium is important.

Solutions:

  1. Incubating Medium
    • Naphtol AS-MX phosphate, di-sodium salt (Sigma) - 5 mg
    • N,N - dimethylformamide - 0.25 ml
    • Fast Blue BB (Sigma) - 30 mg
    • Distilled Water - 25 ml
    • Buffer Solution - 25 ml
    • 10% Magnesium Sulfate Solution. - 2 drops
    • Prepare fresh, shake well and filter before use.
  2. Buffer Solution
    • 0.2M Tris (hydroxymethyl)-aminomethane - 2.4 gm
    • Distilled water - 100 ml
    • Adjust the pH of the buffer to 8.9 with dilute HCl and store at 4°C.
  3. Nuclear Fast Red
    • To 0.2 gm of Nuclear Fast Red add 200 ml of boiling 0.5% aluminum sulfate solution.
    • Keep boiling for 5 - 10 minutes
    • Allow to cool and filter before use.

DOI

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