Re: Dissolving skeletal muscle from bone

From:Kristel Wautier

Hi Ben,

For what it's worth, this method for the clearing of tissues was described
(with some extra add-ins) in 1981 by Hanken & Wassersug. This is the
reference: Hanken, J. & R. Wassersug, 1981. The visible skeleton. A new
double-stain technique reveals the native of the "hard" tissues. Functional
photography, 16: 22-26.
There is also a very similar method described by Taylor & Van Dyke, but I'm
afraid I don't know the details (or exact reference) on that one.


Kristel Wautier
Ghent University
Department of Biology
K.-L. Ledeganckstraat 35
9000 Ghent

Tel.: +32 9 264 52 31
Fax: +32 9 264 53 44

----- Original Message -----
From: "Shelkowsky, Ben" 
To: "Kim Merriam" ; "Histonet"

Sent: Tuesday, December 17, 2002 9:48 PM
Subject: RE: Dissolving skeletal muscle from bone

> Hi Kim,
> Don't know if you received all the information you need from the Histonet,
about dissolving bone, but I'm offering my two cents. When I first got into
histology I'm learned a technique from my mentor in 1965. I was in the army
and he used to make bone preparations of fetuses of various sizes and had
quite an extensive collection. Why he did them I can't remember but they
were beautiful and educational. Anyone who came into the lab and noticed
them, was attracted to them, stared and then were asked a dozen questions
from why and how.
> It's been a long time and as far as I can remember, we immersed the fetus
in a large glass container filled with a dilute KOH (potassium hydroxide).
It's a slow process. After a few days we would change the container with a
fresh batch of KOH. With each change, the muscles, and all the innards would
be dissolving. I guess to some this may sound real ghastly and inhumane.
But... to continue,  this digestion eventually removed all the tissue,
except the skin and bones. The skin was transparent, and the bones were
easily visible. Then we stained the prep with a dilute alizarin red
solution. The stain was taken up by the bone and cartilage.  Lastly  to keep
the prep intact and protect it's integrity, we added glycerol. At this point
the prep is very delicate and it is fragile enough that the bones will
dissassociate from each other.  Shaking the container will cause the
skeleton to break apart and visually it would be a heap of bones at the
bottom of the container. It took about a half dozen changes of glycerin to
remove all the water and make a permanent prep. The glycerin is real thick
and will sort of act as a supportive medium for the bones. Don't shake the
prep as air will be trapped and bubbles are a no no. Always handle gently
throughout . The prep is then permanent and should last for a 100 years.
> For whatever it's worth I hope it helps. We stopped doing this in the
early 80's as it became a controversial procedure with the rise of the
Pro-life movement and late term pregnancies were no longer terminated. When
I went to college I did this technique on rodents and it also worked
> Ben Shelkowsky        ( )
> Community Hospital
> Monterey CA. 93950
> -----Original Message-----
> From: Kim Merriam []
> Sent: Tue 12/10/2002 9:30 AM
> To: Histonet
> Cc:
> Subject: Dissolving skeletal muscle from bone
> Hi all,
> Does anyone know what solution to use to dissolve skeletal
> muscle off of bone, so that the bone remains in tact.  This
> is not for a histology study, but I thought that someone out
> there on the histonet would know how to do this!
> Thanks in advance,
> Kim Merriam
> Cambridge, MA
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