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.
RESULTS
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.