[Histonet] RE: Sanderson's Rapid Bone Stain
sbruce <@t> vetpathservicesinc.com
Thu Feb 19 15:39:59 CST 2009
Here are my responses:
"Suzanne: I no longer use Sanderson's, however we did have nice results on plastic embedded sections, in conjuction with other calcified stains. Use caution in over-staining. I do not recall ever reading about usage on paraffin.
The Villanueva Osteochrome bone stain can also be used on calcified - we used it with fluoro labels, and then viewed using confocal microscopy with beautiful architecture and active new bone formation. Vicki K"
"Sanderson's - we've not used it on wax, but we do use it on 15-20 µm ground sections.
Method for resin:
* Pre-heat Sanderson's to 55-60°C. Stain sections for approx 20 minutes (check every 5 minutes).
* Wash slides in warm water and dry with tissue
* Counterstain with Acid Fuchsin solution for 1 ½ minutues. (2.5g acid fuchsin, 2.5ml glacial acetic acid, 500ml distilled water)
* Drain slides and dry with tissue. "
"I use this stain two ways routinely for "thick" or "ground" sections (35-50 microns) embedded in MMA & DBP. You can stain with SRBS alone @ RT for 3-5 minutes after a brief acid etch in 0.7% FA for 30 sec (rinse in DI water and blot dry) or combine it with a Van Gieson picrofuchsin counterstain for 1-2 minutes (rinse in 100% EtOH and blot dry) after staining first in SRBS @ 60C for 10 minutes (rinse in DI water @ 60C and blot dry).
I have never used or needed to use SRBS on "thin" plastic sections because I routinely use MacNeals tetrachrome combined with Von Kossa and a Modified Goldner's to obtain all my bone related information. With that said, I have also never used this stain on paraffin sections and would be interested in the results if you try it.
"I've used Sanderson's on plastic sections a lot in the past, works well. Recently I tried it on rat tibias, decaled, FFPE. Very disappointing. They look like Toluidine blue-- all shades of blue & nothing else. Now I just stick with von kossa and/or tetrachrome for those.
Suzanne Bruce, R.V.T.
Histologist / Necropsy Coordinator
Vet Path Services, Inc. (VPS)
6450 Castle Dr.
Mason, OH 45040
Lab: (513) 469-0777
Fax: (513) 469-2474
Email: sbruce <@t> vetpathservicesinc.com
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