[Histonet] ATTN: Fabian on MacNeals tetrachrome versus RBS

From:Gayle Callis

What you are looking for is MacNeals Tetrachrome.  I posted the recipe on
Histonet in the last month so go back in Histonet archives for information. 

Also, are you trying to do thick ground/polished bone sections embedded in
methylmethacrylate?  or microtomed PMMA sections? We always 1% formic acid
etched bone sections for 30 sec to 1 min, then rinsed in tap water well,
blotted dry before staining.  This results in better bone components
staining, if you want to see Haversian systems, caniliculi, old and new
osteons, etc. If you do not acid etch with RBS, all you see is red
counterstained bone, and blue to blue green soft tissue components and

RBS or Sandersons rapid bone stain is entirely different than MacNeals,
tintorial differences are apparent after you use these solutions.  One can
also combine a toludine blue staining method (Sterchi and Eurell) with
MacNeals and have a brilliantly stained bone section.    

RBS is a potassim permanganate oxidized methylene blue that results in
production of various other dye components due to the oxidation of the
methylene blue molecule (toluidine blue, azure A, azure B, thionin are some
of the components).   

MacNeals is excellent, in fact, I think far superior for demonstration of
osteoid as compared to RBS.  You can purchase RBS from Surgipath or you can
make up Stevenels blue, these are the same stain solutions, although made
up differently.  I would strongly advise buying RBS, as making Stevenels
blue is a long, tedious and messy process.  You should make up the MacNeals
in house as we found commercial preparations of this stain to be very poor
for bone work.  

How you use the stain depends on how you prepare the section. MacNeals is
stable for years, at least the inhouse solution.  

Hi all,

Does someone has a protocol for Mac Weals tetrachrome? This stain is
supposed to be good in bone staining.

Is there a difference in staining quality between it and RBS (Rapid Bone

Any help will be useful.


Fabien Fuente
Gayle Callis
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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