[Histonet] tooth curl. Long reply

[Rittman, Barry R]

Trisha
It is common for dentin especially to lift up off the slides especially
at the edges.

Four things that you might try
1.	One trick that works is when the sections are being mounted on
slides to brush them out. This is much more gentle than using needles.
Flattened sections on a warming plate are allowed to get almost dry.
Then use a fine camel or sable hair paint brush and gently brush the
dentin of the tooth to eliminate folds or curling. Touch the paintbrush
to a piece of filter paper to eliminate excess moisture that you have
picked up from the section. If the dentin of the tooth in an area dries
before the fold is removed then add just a touch of fluid to that area
and start again.

2.	Paraffin sections of bone and teeth often adhere more
tenaciously to the slide if when the sections are dry to then gently
heat over a small flame until the paraffin just melts. No need to BBQ
them!!

3.	Once slides have been deparaffinized and are in absolute
alcohol, they can be coated with a thin layer of celloidin.  0.5% in
alcohol: ether mixture is the usual solution. The slides are dipped in
the solution for a few seconds, allowed to semi dry until a sheen
appears on the surface and then placed directly into 70% ethanol. This
coating of celloidin is an excellent method to keep sections of tooth
stuck down. 
There are however a few drawbacks. Celloidin is impermeable to enzymes
and so cannot be used for digestion studies. Celloidin is also stained
irreversible by some stains, Celestine blue, alcian blue etc.
If any of these are problems then the celloidinization can be carried
out after staining.
Celloidin is soluble in absolute alcohol. Therefore the final step
before going into the clearing agent should be in alcohol: chloroform
(absolute ethanol and 15% or more chloroform).

4.	Finally. Xylene tends to lift edges of tooth sections and they
often will resist flattening. Instead of xylene can use terpineol. This
is also listed as "oil of lilacin". Terpineol is miscible with absolute
ethanol, xylene and most mountants. It dries very slowly and tissues
remain flexible. The sections are covered with terpineol, drained and
excess is removed by blotting the section with bibulous paper backed
with paper towels. Mountant then applied etc.

Hope that this helps
If you need further details please contact me.
713-500-4134
Barry


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Trisha
Emry
Sent: Wednesday, January 18, 2006 6:38 PM
To: histo
Subject: [Histonet] tooth curl

I just did serial sections through a decaled, paraffin embedded tooth.
I had trouble keeping sections stained with H & E on "plus" slides and
they
often curled up at the edges and folded.

Any advice on how to deal with the ones cut and/or future teeth?

Thanks,
Trisha
U of Washington, Seattle


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