Re: mouse ears

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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 Gayle and all,

I just wanted to point out that the mouse pinnae have only cartilage, but
part of the external ear cannal, middle ear and inner are DO have
calcified bone.  So, it depends on what you are calling "mouse ears".

If you are trying to process adult mouse middle and inner ears, I would
recommend opening the middle ear chamber and the inner ear prior to
fixation and even perfuse the fix through the cochlear turns, if you can.
After that, we use a decalcification of EDTA, on a rotator, 1-2 days
before we dehydrate and imbed- or microdissect.  The EDTA solution we use
is:  291.2 gm EDTA (Sigma # ED4SS-this matters), add to 1L distilled H2O,
adjust pH to 8.0 with approx. 40 ml concentrated acetic acid, once mixed
add distilled H2O for the total volume of 2L.  I use it at room temp.

If you are just working with what most people call "mouse ears", Gayle is
right, you don't need to decalcify.

Karen Pawlowski

On Tue, 16 Feb 1999, Gayle Callis wrote:

> Mouse ears do not contain calcium, just cartilage.  NBF fixation,
> or depending on what you need, and a regular processing schedule
> works fine.  Approx 45 min - 1 hour per change of solvents/paraffin.
>
> Good luck,
> Gayle Callis
>
>




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