Re: mouse feet in paraffin

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From:Taffy <> (by way of histonet)
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Thanks soooo much for your suggestions...we are going to try your technique
next week..I am sure you are right on ALL accounts..I do not think the feet
were left in NBF long enough and I think the dehydration and infiltration
times were too short. I have discussed your protocol with the investigator
and we will try it. I will let you know how it comes out.
Once again..Thanks!!!!  :>)

At 11:00 PM 7/6/00 -0600, you wrote:
>Sorry to disagree, never had problems with decalcified mouse feet in
paraffin but never had much
>success with GMA (poor infiltration and controlling polymerization, PMMA is
a better choice
>here).   Adjustments in fixation, dehydration, and infiltration must be
made to deal with
>1) dense connective tissue on foot pads 2) bones close to each other 3)
skin 4) whole foot
>A challenge but know of other labs who successfully section these tiny
feet.  One lab had a
>series of H&E sections, that looked like footprints walking up a wall, on a
slide hanging like a
>window pane!
>Previous discussion of rat elbows is basically the same technic, works fine.
>Having seen the pitter patter of little mouse feet paraffin sections,
>Gayle Callis
>   ****************************************************
>	Dear Taffy & Monique,
>	When you think about it, sectioning mouse feet from paraffin is never
>going to work.  What you are working with is tiny pieces of bone,
>cartledge, & hardened skin.  They are going to lift out of the paraffin
>every time
>	The only way you are going to get results is to go to resin embedding,
>even then you can expect some problems.  I would suggest something that is
>low viscosity with relatively long infiltration times, I don't think it
>would really matter what type of resin.  However, if you used a hydrophilic
>resin (JB4, LR Gold) & therefore didn't have to do any dehydration, then
>you might get some really great results.
>	Regards
>	Rob W.
>At 02:27 07/05/2000 -0500, you wrote:
>>I have been beating myself up over the SAME problem..the mice feet will NOT
>>section..I thought they were not completely fixed..I have shortened the
>>processing time, I have extended the processing time, we have used EDTA to
>>decal as well as Decal I..nothing seems to work. I keep getting the same
>>kind of sections that you describe. Let me know if you find the answer and I
>>will do likewise.
>>Taffy Hooser
>>At 09:05 AM 6/29/00 -0700, you wrote:
>>>Hello there,
>>>I just joined the histonet so this will be very helpful to me.  I
>>>received two mice feet to embed and cut, they were fixed then I
>>>decalcified them with Cal-ex.  I processed them as usual and blocked
>>>them, but now when I tried to cut them they won't cut, there is just
>>>a hole in the paraffin sections where the sample is.  When I cut it
>>>sounds as if the bone is still a bit hard so I soaked the blocks
>>>again in Cal-ex to see if that would help and I haven't yet tried to
>>>cut them again.  Does anyone esle have any other suggestions that
>>>will help or is there some other trick for cutting bone this is the
>>>first time I have had to do it.  Thanks for any help.
>>>Monique Tourand
>>>University of Alberta
Mary (Taffy) Hooser, HT(ASCP)
Department of Anatomy and Cell Biology
Indiana University School of Medicine
MS-5035, 635 Barnhill Drive
Indianapolis, IN. 46202-5120
Tel: (317)274-2308.
Fax: (317)278-2040.

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