Re: histo. of the Cochlear

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From:Karen S Pawlowski <kna101@utdallas.edu> (by way of histonet)
To:histonet <histonet@magicnet.net>
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Hi Christine,

I routinely do histology on cochlear hair cells.  The hair cells
are often looked at by LM, TEM, and SEM, depending on what is being
investigated.

LM gives a view of presence or absence along with the condition of the
rest of the organ of Corti, often several "turns" of the cochlea can be
viewed in cross-section at one time of the tissue is embedded in paraffin,
JB-4, or frozen.  LM can also be used for a soft tissue prep, where the
organ of Corti is dissected out and mounted so that large "strips" of
organ of Corti can be viewed at one time to look for spots of missing hair
cells.

SEM is used similar to the soft tissue prep to look for more subtle
changes in the stereocilla at the apical ends of the hair cells.
TEM is the best way to look for cytoplasmic changes, but of course you can
only look at a few hair cells at a time.  I have used medcast, araldite
and Spurr's for this.

As for decalcification, you can do TEM, SEM, and LM without it, but that
gets pretty tricky.  When I decalcifiy, I use an EDTA mixture for a day or
two.  The recipe I use for this now works faster, with no difference in
arcitecture from the recipe I used to use.  Here is the recipe:

291.1 gm EDTA (Sigma #ED4SS)
1 L distilled water
mix and add about 42 ml of acetic acid to bring pH to 8.0
Bring total volume to 2 L

I decalcify at room temp.

If you need more help.  You can e-mail me directly, kna101@utdallas.edu

Karen Pawlowski
Sr. Research Assoc.
UT Southwestern Med. Ctr., Dallas/
PhD Candidate
UT Dallas

On Mon, 8 Feb 1999, Christine Lee wrote:

> I have been asked about histology of the cochlear of the ear by a student
> who wants to study the hair cells. my advice to him has been that he will
> most probably have to do it at the ultrastructural level using TEM. i do't
> think that light microscopy will help much. Has anyone any experience with
> this sort of thing?. I have no none. I think the specimen would need
> probably need decalcification. What would decal. do to the ultrastructure of
> the hair cells???
>
>
> Christine Lee,
> Senior Scientific officer,
> Veterinary Pathology and Anatomy,
> University of Queensland.
> C.lee@mailbox.uq.edu.au
>
>
>




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