Giemsa Stain for Glycol Methacrylate Sections

Technovit Glycol Methacrylate: Technical Information and Help From Heraeus Kulzer.

This Giemsa stain works well for a variety of tissues.

Procedure:

  1. Stain in Giemsa working solution at room temperature for 1.5 hours.
  2. Rinse in dilute acetic acid (4 drops in 100 ml. DI water) for 2 seconds.
  3. Dip in 96% alcohol.
  4. Dip in 96% alcohol.
  5. Rinse in isopropyl alcohol 3 times for 2 minutes each.
  6. Clear in xylene and mount.

Results:

Nuclei - violet
Cytoplasm - blue
Erythrocyten - pin
k

Solutions:

Giemsa Solution
  • 0.15 g Azure A
  • 0.30 g Methylene Blue
  • 0.36 g. Eosin Y
  • 0.04 g Phloxine B
  • 60 ml Glycerine
  • 40 ml Methanol
  • Add dyes to glycerine and methanol, mix well. Incubate overnight in 55 - 58C oven stirring occasionally. Stable for a year.

Giemsa Working Solution

  • 4 ml Giemsa Stock Solution
  • 40 ml phosphate buffer solution

Phosphate Buffer Solution

  • 7.25 g Sodium phosphate, monobasic
  • 2.75 g Sodium phosphate, dibasic
  • 1000 ml Distilled water
  • 10 ml 10% Triton X-100
    Store at 4C.

Citation:

Gerrits, P. O. and Smid, L., "Staining Procedures for Tissues Embedded in 2-Hydroxyethyl Methacrylate", Heraeus Kulzer.


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