Re: Spalteholtz Solution

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From:"J. A. Kiernan" <> (by way of histonet)
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On Mon, 29 Nov 1999, Jay Turner wrote:

> For demonstrating the intraosseous blood supply to the distal femur, I have
> come across several protocols that inject with india ink and immerse with
> "Spateholtz solution" to make the tissue transparent.  Is anyone familiar
> with this technique?  What is Spalteholtz solution and where can it be
> purchased?

   It's a mixture of equal volumes of benzyl benzoate and
   methyl salicylate. In the traditional clearing method
   the formaldehyde-fixed object is bleached in hydrogen
   peroxide, then  _completely_ dehydrated (alcohols),
   and then passed through changes of benzene and benzyl
   benzoate into the mixture. Specimens must be kept in
   glass jars because the solvents attack polymethacrylate
   plastics. The whole process includes 12 14-day steps.
   [CFA Culling 1974. Handbook of Histopathological and
   Histochemical Techniques (including museum techniques),
   3rd ed. London: Butterworth. pp.550-551.]

   Benzyl benzoate and methyl salicylate can be bought
   from any ordinary chemical supply house.

   John A. Kiernan,
   Department of Anatomy & Cell Biology,
   The University of Western Ontario,
   LONDON,  Canada  N6A 5C1

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