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Electron Microscopy Sciences

Technical Data Sheets

Price's Giemsa Stain Method

EMS Catalog #26156

Fixation

10% Buffered Neutral Formalin or Zenker's. Use no fixation technique that will destroy erythrocytes.

Sections

Paraffin @ 4 microns

Staining

  1. Deparaffinize with two changes at 2 minutes each in xylene. Rinse in absolute alcohol for two changes, 2 minutes each.
  2. Rinse in 95% Alcohol for two changes after a one minute immersion. If necessary to further remove mercuric chloride crystals, use an Alcoholic Iodine Solution which will be further removed in successive alcohols.
  3. Rinse through 80%, 70%, 50%, alcohols for one change, 1 minute each.
  4. Rinse in distilled water for 15 seconds.
  5. Buffer in the Phosphate Buffer Solution, for 30 minutes.
  6. Stain in the Giemsa Working overnight. To prepare Giemsa Working:

    •  Geimsa Stock .......3 parts
    •  Phosphate Buffer pH 7.0 ..97 parts
  7. Rinse in the buffer solution.
  8. Rinse in Acetic Acid, 0.2%, for one minute, absolute alcohol for two changes, 15 seconds each. This step should take only 90 seconds!!!
  9. Dip in xylene for two changes, two minutes each.
  10. Mount in Permount.

Cells

Blue Malarial parasites
Blue Tissue Nuclei
Dark Blue Bacteria
Black Malarial Pigment
Blue Schistosomic egg shells
Pale Pink Collagen, etc.
Pink-Rose Erthrocytes

 

References

AFIP Manual of Histological Staining Methods , 3 rd ed., L.Luna:

Product Information

Giemsa Stock Solution
Phosphate Buffer
Acetic Acid
Alcoholic Iodine
Price's Giemsa Stain Method