[Histonet] storage of frozen cryosections (again)

[Kathleen Spencer]

We pick up our sections and put them in a slide box that is in the 
cryostat and keep them there until we are finished sectioning. Then we 
rush them to the -80 freezer. We put a dessicant pack or two in the box 
with the slides, and we wrap the slide box in foil, then stick that 
into a plastic bag that we label as to what the slides are. When we are 
ready to do the next step,  we remove the box from the freezer, quickly 
take out the slides we need & put the box back. We then place a warm 
finger under the section for half a second, then put the slide into 
cold fix or 70% alcohol, just depends on what we are using the slides 
for. It is usually LCM for RNA. but it could work for other things like 
IHC. The morphology is terrible on fresh frozen tissue sections, so for 
IHC we always perfuse the animal with PFA and store the tissue in in 
20% sucrose, then freeze the tissue for cryosections. Then they can air 
dry because they are fixed.
Make sense? I hope this helps.

Kathleen Spencer HT (ASCP)
Lab Manager/LCM Supervisor
UTHSC


On Feb 2, 2005, at 9:53 AM, Anna Elisse Beaudin wrote:

> Dear Histonet,
>
>    I have a question regarding the storage and preservation of frozen
> cryosections.  I apologize if I have asked this question before, but
> I'm still unclear as to the best approach.  Currently, when I collect
> cryosections, I dry them overnight at room temp and than use them
> immediately the next day.  however, I would like to be able to collect
> sections to use at later timepoints.  My concern is that O/N drying at
> room temperature followed by freezing and storage at -20 or -80 might
> cause ice crystals or otherwise affect the quality and morphology of my
> sections.  Additionally, even if I were to freeze the sections
> immediately following sectioning, they are still collected on slides at
> room temperature, and thus I wonder if I would have the same problem
> with thaw/freeze.  I would greatly appreciate anyone's advice with
> this... I would really like to be able to preserve more sections for
> later use!
>
> Thanks so much in advance!
> Anna Beaudin
> Division of Nutritional Sciences
> Cornell University
>
>
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