[Histonet] MacNeals Tetrachrome stain solution recipe

[Gayle Callis]

Dear LaCinda,

Here is the recipe you are looking for.  I have stained PMMA embedded bone
sections, thicker/ground with this, but I did use 1% formic acid etching
for 30 seconds in ultrasonicator, a very good water rinse after this, then
dry section and immerse.  This stain can also be combined with Toluidine
Blue, Sterchi method for even more brilliant results particularly if you
want to demonstrate ostoid!  


MACNEAL'S TETRACHROME

STOCK SOLUTION
	0.5g methylene blue
	0.8g azure A eosinate
	0.1g methylene violet CI# 52041
	250 ml methanol
	250 glycerin
	Stir, leave at 50C for 12 hours, then 37C for 3 days.  Filter, store in
dark. It is good for up to 	years!  Older solutions stain very well, if
not better. 
WORKING STAINING SOLUTION
	5 mls stock stain solution, 95mls distilled water


Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology 
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)