[Histonet] RE:Sanderson's RBS image

[Suzanne Bruce]

Thanks Gayle,
 
I also looked at the Surgipath image, my slide was much more brilliant then that one.  I did use the acid fuchsin counter stain.  I just wasn't sure if mine would be considered 'over stained.'  I'll send you an image that Jack Ratliff sent me that compares to what I was seeing today.  I'm not using GMA, but Technovit 7200 ~100 micron thick ground sections.
 
Suzanne
 
Date: Thu, 19 Mar 2009 10:38:51 -0600
From: "gayle callis" <gayle.callis <@t> bresnan.net>
Subject: [Histonet] RE: Sandersons rapid bone stain photo
To: "'Histonet'" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000401c9a8b1$30e43630$92aca290$@callis <@t> bresnan.net>
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On looking at the Surgipath photo, the bone is very pale although the
osteocytes, osteoid and soft tissues look good.  I am not sure what
counterstain (two are touted in the stain data sheet) was used as seen in
this photo.   One is a basic fuchsin that turns the bone red.  Be careful of
overstaining with fuchsin counterstain.  The other is light or fast green
and since we never used that, I can't comment on the color contrast with
bone and soft tissue components.  This may be the stain used in
photograph???



It always is a good idea to look at the stain while performing the actual
staining method.  After prescribed or desired time in RBS, simply rinse
quickly with running hot tap water,  blot and examine under the microscope
to see depth of stain for soft tissue/bone components. To view a dry
section, simply place a coverglass on top of section for higher power
examination.   If components stained by RBS not dark enough, you can return
to the hot RBS.  Rinse then blot, and counterstain after that.  It does not
hurt the bone section to dry.  In fact we preferred a dry section when going
back into either stain just to prevent dilution of the stain solution,
particularly when doing hand ground thick section surface staining. 



Gayle Callis

HTL(ASCP)HT,MT

Bozeman MT 59715