[Nhcoll-l] Clearing & Staining issue

[Sebastien Enault]

Dear Simon,
Thanks a lot for your detailled reply. I suspected some form of fat tissue
as well, and these are indeed captive bred foetal rats (in 20%KOH/glycerine
on the pictures). I was hesitant to use acetone once they are stained, but
next time I have to stain further specimens I'll give it a try.
Thanks again and all the best,
Sebastien

======================
Sebastien Enault, PhD
www.kraniata.com

On Tue, Apr 24, 2018 at 6:46 PM, Simon Moore <couteaufin at btinternet.com>
wrote:

> Hi Sebastian,
>
>
>
> These look like mice and preserved in glycerine or propylene glycol (I use
> the former).  The white tissue looks to me like fatty deposits, mainly
> tri-glyceride.  This will become more transparent if the concentration of
> the glycerine is increased to about 70/80%.  I have found that lab. Bred
> mice tend to have more fat bodies than wild ones.  In some cases, I
> skinned, eviscerated and fixed the specimens (10% formalin), then
> dehydrated them into acetone to reduce the body fat problem, then
> macerated, stained and preserved in 10% glycerine, gradually increasing the
> concentration of glycerine to c. 80%.
>
> For foetal mice, that need no skinning, the process is much the same.
> However, as I first stated, the more concentrated glycerine used reduces
> the opacity of the fat bodies.
>
>
>
> With all good wishes, Simon.
>
> Simon Moore, MIScT, FLS, RScI, ACR,
> Conservator of Natural Sciences and Cutlery Historian.
>
> www.natural-history-conservation.com
> Sent from Mail for Windows 10
>
>
>
> *From: *Sebastien Enault <voraghl at gmail.com>
> *Sent: *24 April 2018 15:37
> *To: *nhcoll-l at mailman.yale.edu
> *Subject: *[Nhcoll-l] Clearing & Staining issue
>
>
>
> Hello all,
>
> I have a quick question for those of you that routinely use clearing and
> staining for embryonic specimens. I mostly use the technique on small fish
> and herps without issues, but when processing foetal/juvenile rodents, I've
> had white tissue not clearing at all during the process. Attached are a
> couple of pictures to illustrate the issue. This happened on two distinct
> batches, and if any of you know what's going on and how to solve the issue,
> that would be greatly appreciated. So far the only way it could be removed
> is by careful manual scraping, which is obviously not satisfactory
> considering how time consuming it is and due to the increased risk to
> damage such fragile specimens.
>
> Thanks and best regards,
>
> S. Enault
>
> ======================
> Sebastien Enault, PhD
> www.kraniata.com
> <https://urldefense.proofpoint.com/v2/url?u=http-3A__www.kraniata.com&d=DwMFaQ&c=cjytLXgP8ixuoHflwc-poQ&r=LpYc_Z_iN1KRw0hheb3x6-8MJUMu482qfHowpGYJqwc&m=6PM7vVYv8TDiqCfeB7kk6JJd6BlQJM3lTi2RtGaI_kM&s=was9EyF54MNYgSpeG-aMUShaO8kOYfsmCUlgPUo3F5w&e=>
>
>
>
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