Yellowsolve I
for Fibrin

Solutions

Optional – an elastic fibre stain: Orcein or Weigert's iron resorcin fuchsin, or variant.
Celestine blue-hemalum nuclear stain, or similar.
Trichlorethylene
Tetrachlorethylene
Solution A
Phloxine B 0.5 g
Calcium chloride 0.5 g
Distilled water 100 mL
Solution B
Lissamine flavine FF 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.

Method

  1. Dewax sections with xylene.
  2. Place in trichlorethylene for 24 to 48 hours.
  3. Bring sections to water.
  4. Stain elastic fibres if desired.
  5. Stain nuclei with the celestine blue-hemalum sequence.
  6. Wash well with water.
  7. Place in solution A for 30 minutes.
  8. Rinse with 2-ethoxyethanol then with tetrachlorethylene.
  9. Place into tetrachlorethylene overnight.
  10. Rinse with 2-ethoxyethanol.
  11. Differentiate with solution B, controlling microscopically.
  12. Rinse with 2-ethoxyethanol.
  13. Clear in xylene and mount with a synthetic resinous medium.

Expected results

Notes

  1. 2-Ethoxyethanol is also known as cellosolve or ethylene glycol monoethyl ether, and has the formula CH3CH2OCH2CH2OH.
  2. Tetrachlorethylene has the formula Cl2C=CCl2 or C2Cl4
  3. Trichlorethylene has the formula ClHC=CCl2 or C2HCl3
  4. Both trichlorethylene and tetrachlorethylene 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.

 


 
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Last updated January 2010.

 

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