Hi Paula,
I am cutting at -24, but have tried going as warm as -18. I am
currently learning with 30uM sections with the ultimate goal of moving
towards 5 or 10uM. Our lab standard has been collecting into millonigs
solution and doing free floating IHC. We generally have no issue with
this technique, but do lose some of the peripheral damaged tissue near
the infarct in our stroke brains. We're trying to work up painting the
samples directly onto slides and skipping free floating staining to get
a better end product.
My current samples are very old, they were collected into 4%PFA in 2017
and cryoprotected by freezing in OCT cryomatrix in a little dish
floating on LN2 in 2019. They've been stored at -80 since then. Usually
we process the brains within a few weeks of collecting them so this
particular tissue isn't our ideal situation. We're using old tissue to
practice technique, so the current samples aren't going to be used for
any actual analysis. When we heard about the tape method of collecting
we were very curious as the the opinion other labs have about it. Have
you tried it?
Thanks for the answer!
Heather
Heather Deziel, MSc.
Laboratory Technician, CNS|CRO 550 University Ave, Charlottetown, PE C1A
4P3
(a subsidiary of Neurodyn Life Sciences Inc.)
On 2021-04-15 10:29, Patpxs wrote:
> Good Morning Heather,
>
> I have some questions about how you cut frozen brains.
>
> What temperature are you cutting at?
> How thick are your sections?
> How are your samples frozen? Flash freezing, slow freezing, iso-pentane in
> LN2?
>
> Your answers may provide clues to help you get better cryosections.
>
> Paula
>
> Sent from my iPhone
>
>> On Apr 15, 2021, at 5:39 AM, Heather Deziel via Histonet
>> wrote:
>>
>> Hello Histonet,
>>
>> I'm looking into working up a tape transfer method of collecting
>> cryosections of brain while preserving infarct to be used in IHC. I
>> find that when I try and section heavily damaged regions of the brain
>> the tissue tears and and I lose it. Has anyone got any recommendations
>> about the the Section-lab transfer tape (Kawamoto method), using the
>> circuit plating tape recommended here
>> (https://www.future-science.com/doi/full/10.2144/btn-2018-0021) or the
>> cryojane system from Leica?
>>
>> Thank you,
>>
>> Heather
>>
>> Heather Deziel, MSc.
>>
>> Laboratory Technician, CNS|CRO 550 University Ave, Charlottetown, PE C1A
>> 4P3
>>
>> (a subsidiary of Neurodyn Life Sciences Inc.)
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