Re: tissue embedding for microtomy


"Gary G" <see.signature@bottom> wrote in message
news:o1anu21lg44oco6ulenr38jpslcbqvf6l8@xxxxxxxxxx
On Sun, 04 Mar 2007 23:59:37 -0500, "Harry Busk" <hbusk@xxxxxxxx>
wrote:

On Sun, 04 Mar 2007 09:08:07 -0500, Kevin Cunningham
<smskjv@xxxxxxxxxxxxxx> wrote:

[]snip for brievety



IMO, isopropanol will never work since by definition, it is 30% water.
This is a killer right from the start.

Consider HMDS and ethyl alcohol. These are for dehydration. There
are other agents that will dehydrate. After embedding, then you face
the slicing. That is normally done with Diatome instruments.

I do not know of a simple way to do this. If there is, I would indeed
like to know.

gary g.


Kiss French. Drink California.

I'm affraid I have to contradict Gordon on this one (with all due respect):
dehydration and the use of IPA as an intermedium between ETOH and paraffin
is mainstream microtechnique, in use since a long time. I've used it since
more that 15 years and it works very well for both animal and plant tissues.
(I used IPA reagent grade, at the time from Janssen Chemicals, now Acros
Chemicals).

The technique is described in lots of manuals, amongst them Peter Böck:
"Romeis' Mikroskopische Technik", 17 ed., 1989 (p.127-128) and I think (have
to look that up, but frankly: don't know where that book hangs out) that
Romeis mentioned it in the 16th edition (1968) too.

Böck advises to use paraplast after the IPA, but I have infiltrated tissues
after IPA with about every paraffin one can imagine, including home made
"improved" household paraffin containing beeswax, home made "improved"
household paraffin containing .5% dimethylsulfoxid and more of those eh...
astrological and alchemical brews.

Yvan.




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