[Histonet] stains for visualizing new bone growth

[Jack Ratliff]

Robin, I typically stain first with Von Kossa and then counter with MacNeal's. This provides a very nice contrast where obviously mature mineralized bone is black and newly formed bone (osteoid) is grayish-green color. Additionally, your marrow space is nicely contrasted with clear visualization of osteoblasts and osteoclasts lining the bone surface. At the microscope this is a one-stop-shop stain for collecting static bone histomorphometry.

Another nice contrasting stain is a modified Goldner's trichrome stain. With this stain cell nuclei are stained first with a Weigert's (iron) hematoxylin, then newly formed bone (osteoid) is stained red with an acid fuchsin/ponceau stain, next an orange G cytoplasmic stain covers the rest and a light green SF yellowish stain follows up with a nice green contrast of the mineralized bone. Very clear differentiation between mineralized bone (green) and newly formed bone (red). This stains works very well with auto threshold functions on some histomorph systems as it has a very nice contrast for the software to recognize.

Of course Masson's trichrome works as well but it is typically used on decalcified paraffin embedded sections. I have found the Masson's staining kit at Sigma-Aldrich and kits for all the other stains mentioned can be found at Dorn and Hart Microedge. In fact, Dorn and Hart Microedge (www.dornandhart.com) has a lot to offer now with regards to mineralized bone (hard tissue with or without implant materials) and resin embedded histology.

Good luck to you and let me know if you have any additional questions. I would also be happy to share images with you if interested.

Jack

On Feb 1, 2011, at 1:44 PM, "Robin Dean" <robin_dean <@t> compbio.com> wrote:

> Does anyone know of a good stain to use to clearly show new bone growth
> other than von Kossa stain?
> 
> Would appreciate any suggestions anyone might have.
> 
> 
> 
> Thank you,
> 
> 
> 
> Robin
> 
> Robin R. Dean, Ph.D.
> 
> Senior Scientist & Study Director
> 
> Comparative Biosciences, Inc.
> 
> 786 Lucerne Dr.
> 
> Sunnyvale, CA
> 
> (408) 738-8060
> 
> robin_dean <@t> compbio.com
> 
> 
> 
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