Protocol
McNamara's Giemsa
General Oversight Stain
Materials
- Stock Giemsa
- Working Giemsa
Material Amount Stock Giemsa 10 mL Acetone 10 mL Methanol 10 mL Distilled water 100 mL Make the working solution just before using.
- Colophonium solution
Material Amount Colophonium 15 g Absolute ethanol 100 mL - Rinse solution
Material Amount Acetone 70 mL Xylene 30 mL
Tissue Sample
Helly or neutral buffered formalin fixed tissue are recommended. Thin (3 µ) sections are desirable.
Protocol
- Bring sections to water with xylene and ethanol.
- Rinse well with distilled water.
- Place into working Giemsa for 10 to 15 minutes.
- Rinse with tap water water and check staining microscopically.
- Differentiate with colophonium solution until colour contrast is satisfactory. Control microscopically. If a film develops renew the colophonium solution.
- Rinse several times with rinse solution. Do not allow to dry.
- Rinse several times with xylene. Do not allow to dry.
- Mount with a synthetic resinous medium.
Expected Results
- Nuclei – blue-purple
- Cytoplasm – pink
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Putt, F.A., (1972)
Manual of histopathological staining methods
John Wiley & Sons, New York, NY, USA
