McClung's Jenner-Giemsa
for Malaria Parasites & General Oversight
Materials
- Stock Giemsa
- Stock Jenner
- Working Jenner
Material Amount Stock Jenner 1 volume Distilled water 1 volume Make the diluted solution just before using.
- Working Giemsa
Material Amount Stock Giemsa 1 volume Distilled water 10 volumes Make the diluted solution just before using.
- Dehydration Fluids
Fluid Amount of Acetone Amount of Xylene 5% Xylene in Acetone 95 volumes 5 volumes 30% Xylene in Acetone 70 volumes 30 volumes Equal parts Xylene and Acetone 50 volumes 50 volumes 30% Acetone in Xylene 30 volumes 70 volumes 5% Acetone in Xylene 5 volumes 95 volumes
Tissue Sample
A fixative containing mercuric chloride and ethanol is recommended, but many fixatives may be used, although staining results may vary. 3µ paraffin sections of neutral buffered formalin fixed tissue are usually suitable.
Protocol
- Bring sections to water with xylene and ethanol.
- Remove mercury pigment if necessary.
- Rinse well with distilled water.
- Place into working Jenner for 4 minutes.
- Without rinsing, place into working Giemsa for 15-20 minutes.
- Rinse well with distilled water to differentiate.
- Dehydrate with the dehydratiion fluids in the order given above.
- Clear with xylene and mount with a synthetic resinous medium.
Expected Results
- Nuclei of tissue – blue-purple blue
- Cytoplasm of tissue – pink
- Nuclei of parasite – red-purple red
- Cytoplasm of parasite – blue
- Schüffner’s stippling – red
Notes
- If the sections are too blue, dilute acetic acid in distilled water may be used to remove the excess (1% or less). Wash with distilled water afterwards to remove residual acid.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Humason, G.L., (1967)
Animal Tissue Techniques
W.H. Freeman & Co., San Francisco, CA, USA
