Modified Yellowsolve
for General Oversight

Solutions

Hemalum

Trichlorethylene

Solution A
Phloxine B	0.5	g
Calcium chloride	0.5	g
Distilled water	100	mL

Solution B
Luxine pure yellow	to	saturation
2-Ethoxyethanol	50	mL
Ethyl phosphate	50	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.

Lendrum says, "If … the time of differentiation is kept short the result is comparable to the best examples of the old Masson's erythrosin-saffron technique." Masson's erythrosin-saffron is the same technique as the HPS (hematoxylin-phloxine-saffron) using phloxine instead of erythrosin.

Method

1. Dewax sections with xylene.
2. Place in trichlorethylene for 24 to 48 hours.
3. Bring sections to water.
4. Stain nuclei with hemalum, differentiate and blue.
5. Wash well with water.
6. Place in solution A for 30 minutes.
7. Rinse with 2-ethoxyethanol.
8. Differentiate with solution B, controlling microscopically, until collagen is yellow.
9. Rinse with 2-ethoxyethanol.
10. Clear in xylene and mount with a synthetic resinous medium.

Expected results

• Nuclei  –  blue
• Muscle  –  red
• Background  –  yellow

Notes

1. Stop differentiation when collagen is yellow and muscle is still red.
2. 2-Ethoxyethanol is also known as cellosolve or ethylene glycol monoethyl ether, and has the formula CH₃CH₂OCH₂CH₂OH.
3. Trichlorethylene has the formula ClHC=CCl₂ or C₂HCl₃
4. Trichlorethylene should be used in a fume hood.

 

Reference
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.

Translate in Google Translate Instructions