Re: frozen sections revisited

From:Ryan Blair Rountree


I cut my sections and let them air dry.  I use Superfrost plus slides from
Fisher - they are just about the best of any I've tried for stickyness.
ProbeON Plus slides by fisher are a little stickier, but they have a large
painted white patch that is so annoying I don't usually use them.

If I'm storing them for RNA in situs later, I let them dry 1st, put my
slide box into tupperware with Drierite sprinkled in the bottom of the
tupperware, close it up and freeze it at -80.

When they are ready to use, I pull them out and let them warm up in the
tupperware for 1-2 hours before use.  The key seems to be not letting the
slides get wet from condensation and then everything works great.


On Sun, 27 Oct 2002, wrote:

> Histonetters,
> I know this subject has been done on HISTONET before, but can we open this
> up again.  how do you process frozen sections to get the best slide
> adhesion?
> do you fix immediately in cold fix in the cryostat or airdry and then fix?
> what do you fix in, ie cold acetone,etc.?  do you always try to stain
> slides soon after preparing?  do you store FS for staining later?  how are
> FS stored, ?wrapped individually in foil and plastic bags, etc.?  do you
> store them unfixed or fixed?  how are stored FS retrieved from the freezer?
> Thanks for responding to this mini survey.
> Patsy Ruegg
> James,
> are you talking about cells in suspension or tissue sections lifting off
> silane coated slides?
> for keeping FS on plus slides, i pick them up and let them airdry well for
> at least 10 min. at RT.  there are some different methods for what to do
> after drying to fix and stain right away or store frozen unfixed until
> ready to stain, but i think the initial drying at RT goes a long way to
> keep them on the slides.  some people have a jar of cold fixative in the
> cryostat and fix immediately after sectioning in the cryostat and then air
> dry, but i have had trouble with preservation of morphology doing it this
> way.  people have their favorite methods that work the best in their hands.
> my WS partner said she had some antigenicity loss from storing dryed
> unfixed sections in the freezer, but i have always done it that way with no
> apparent problems.
> Original Message:
> -----------------
> From:
> Date: Fri, 25 Oct 2002 11:52:45 -0400
> To:
> Subject:
> Has anyone had any problems with lifting off cells for LCM that have been
> picked up on SuperFrost Plus Slides?  How
> did you resolve the problem?
> Also does anyone have any suggestions for keeping fresh frozen cyrostat
> sections on any kind of slide.
>                          Thanks,
>                              JPZ
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Ryan Rountree
Department of Developmental Biology
B301, Beckman center
279 Campus Drive
Stanford, CA 94305
(650) 725-7599
Fax: (650) 725-7739

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