148 Connective Tissue (chap. 13) 



procedure: 



1. Deparaffinize and hydrate slides to water; remove HgCl2. If HgCU 

 is absent from fixative, mordant in saturated aqueous HgClo, plus 

 5% glacial acetic acid: 10 minutes. Wash, treat with Lugol's and 

 sodium thiosulfate, wash and rinse in distilled water. 



2. Stain in Mallory I: 15 seconds. 



3. Rinse in distilled water: 10 or more seconds, to differentiate reds. 



4. Treat with phosphomolybdic acid: 1-5 minutes. 



5. Rinse briefly in distilled water. 



6. Stain in Mallory II: 2 minutes. 



7. Rinse in distilled water. 



8. Differentiate aniline blue in 90% ethyl alcohol. 



9. Dehydrate in absolute alcohol, clear and mount. 



results: 



nuclei — red 



muscle and some cytoplasmic elements — red to orange 



nervous system — lilac 



collagen — dark blue 



mucus, connective tissue and hyaline substance — blue 



chitin — red 



yolk — yellow to orange 



myelin and red blood cells — yellows and orange 



dense cellular tissue (liver) — pink with red nuclei 



bone matrix — red 



comments: 



From distilled water (step 7) the slides can be run through the "up" 

 series of alcohols, thereby controlling the blue color. Aqueous-alco- 

 holic solutions differentiate the amount of Mallory II left in the 

 various parts of the tissue. After the slides have remained in the ab- 

 solute alcohol for a couple of minutes they should have changed from 

 a muddy purple to a clear blue and red. An acetic acid rinse, as men- 

 tioned (page 147), following step 7 contributes to the transparency 

 of the sections. 



For better nuclear detail, Mallory I can be preceded by alum hema- 

 toxylin staining. (Results: nuclei — blue) 



Landrum and McFarlane (19-^0) recommend addition of celestin 

 blue. Follow step 1 with: 



