Sanderson's Rapid Bone Stain

From:Cathy Mayton

Dear Karen,
A friend of mine saw your posting on Histonet (I get the digest so I am a day behind) in regards to the Sanderson's Rapid Bone Stain (RBS).  I am assuming that you are wanting to stain mineralized bone sections.  This is probably not exactly as the Surgipath instructions but this is how I do it.  The stain is very easy to do and is reproducible.  I think you will like the stain.
1.  Preheat the RBS to 55-60 degrees in a water bath or oven. 
2.  Stain for 30 seconds to 2 minutes in the RBS
3.  Rinse stained sections in warm tap water to remove excess stain and dry section with a laboratory wipe.
4.  Counterstain with either room temperature acidified acid fuchsin (found in the green AFIP manual) for 10-15 seconds, drain the stain off the section and then wipe the section with a laboratory wipe.  Or room temperature van Gieson for 2 minutes, dehydrate quickly through 95%-100% ETOH and wipe section dry.  If the bone is fluorochrome labeled use the acid fuchsin.
Acid fuchsin counterstain
osteiod                             blue
cells within marrow            blue
soft tissues                       blue
mineralized bone               pink
van Gieson counterstain
osteoid                              blue-green
cells within marrow             blue
soft tissues                        blue-green
mineralized bone                yellow-orange
The stain is very easy to do and is reproducible.  I think you will like the stain.  I "discovered" the stain while working in the Bone & Joint Research Lab in Salt Lake City under the direction of Roy D. Bloebaum, Ph.D.  We needed a stain that worked better than what was currently available.  The stain clearly differentiates mineralized bone from the non-mineralized tissues.  We also found that the RBS will differentiate bone that is dying and demineralizing.  This bone will look purplish if you use the acid fuchsin.  There is a previous publication that Dr. Kent Bachus and I co-authored a few years ago that was published in the Journal of Histotechnology.  This paper used back scatter electron imaging to confirm that the purplish areas was demineralized bone.  The publication is listed on my web-site if you are interested.
GLP Compliant Laboratory
Cathy A. Mayton
Project Director
Wasatch Histo Consultants, Inc.


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