A word of advice, make up your own stain solution.  We and another lab via
private discussion found commercial preparation of this stain were very
poor for PMMA bone section staining. 

MACNEALS TETRACHROME

	STOCK SOLUTION

	0.5g methylene blue
	0.8g azure a eosinate
	0.1g methylene violet
	250 ml methanol
	250 glycerin
	Stir, leave at 50C for 12 hours, then 37C for 3 days.  Filter, 	store in
dark.

WORKING STAINING SOLUTION

5 mls stock stain solution, 95mls distilled water.  Bone is 1% formic acid
etched for 30 sec to 1 minute, rinsed well with water before staining. Blot
section before immersing in this stain solution.  You can combine this
stain with Toluidine blue method (Eurell and Sterchi) for even more
brilliant results.  


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)



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