Yellowsolve II
for Cellular Inclusions
This method is also known as the Rhoda-Coomassie.
Materials
- Celestine blue-hemalum nuclear stain, or similar.
- Trichlorethylene
- Solution A
Material Amount Lissamine rhodamine B 1 g Acetic acid, glacial 2.5 mL Distilled water 97.5 mL - Solution B
Material Amount Coomassie fast yellow G to saturation 2-Ethoxyethanol 100 mL
Tissue Sample
5µ paraffin sections of neutral buffered formalin fixed tissue are likely suitable. It should be noted, however, that the authors of this method favoured extended fixation in formal sublimate of up to 10 days for fibrin. This fixative is now deprecated due to its mercuric chloride content. With other staining methods, pretreatment of sections with Bouin’s fluid for an hour at 60°C can compensate for the lack of mercury fixation to a large degree.
Protocol
- Dewax sections with xylene.
- Place in trichlorethylene for 24 to 48 hours.
- Bring sections to water.
- Stain nuclei with the celestine blue-hemalum sequence.
- Wash well with water.
- Place in solution A for 5 minutes.
- Rinse with 2-ethoxyethanol.
- Differentiate with solution B, controlling microscopically.
- Rinse with 2-ethoxyethanol.
- Clear in xylene and mount with a synthetic resinous medium.
Expected Results
- Cell inclusions – red
- Background – yellow
- Nuclei – black
Notes
- Inclusions stained may include mast cells, plasma cells and Paneth cells as well as fibrin.
- 2-Ethoxyethanol is also known as cellosolve or ethylene glycol monoethyl ether, and has the formula CH3CH2OCH2CH2OH.
- Trichlorethylene has the formula ClHC=CCl2 or C2HCl3
- Trichlorethylene should be used in a fume hood.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Lendrum A C, Fraser D S, Slidders W and Henderson R. (1962)
Studies on the character and staining of fibrin.
Journal of clinical pathology, v. 15, p. 401.
