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 - pink

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 - 58°C 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 4°C.

Citation:

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