Protocol
Best's Carmine
for Glycogen
Materials
- Mayer’s hemalum or a similar nuclear stain
- Stock Solution
Material Amount Carmine 2.5 g Potassium carbonate 1.2 g Ammonia 25 mL Distilled water 75 mL Combine the water, potassium carbonate and carmine. Boil for 15 minutes. Cool and filter. Add the ammonia.
- Working solution
Material Amount Stock solution 25 mL Methanol, absolute 35 mL Ammonia 35 mL - Differentiator
Material Amount Ethanol, 95% 40 mL Methanol, absolute 20 mL Distilled water 50 mL
Tissue Sample
5µ paraffin sections of neutral buffered formalin fixed tissue are suitable. It is often recommended that an alcoholic fixative be used to preserve glycogen, but the majority may be demonstrated following formalin fixation. Alcohol fixation is not usually required except for critical applications.
Protocol
- Bring sections to water via xylene and ethanol.
- Stain nuclei with hemalum. It is not necessary to blue.
- Rinse with distilled water.
- Place in the working solution overnight.
- Rinse well with water.
- Place in the differentiator until glycogen granules are prominent.
- Dehydrate with ethanol.
- Clear with xylene and mount with a resinous medium.
Expected Results
- Glycogen – red
- Nuclei – blue
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Gray, Peter. (1954)
The Microtomist’s Formulary and Guide.
Originally published by: The Blakiston Co.
Republished by: Robert E. Krieger Publishing Co.
Citing:
Best, (1906)
Archiv für mikroskopische Anatomie, v. 23, p. 520
Bonn, Germany - Bancroft, J. D. and Stevens, A., (1977)
Theory and practice of histological techniques
Churchill Livingstone, Edinburgh, UK
