[Histonet] storing sections
Geoff McAuliffe
mcauliff <@t> umdnj.edu
Thu Apr 15 15:19:00 CDT 2004
Hi Katy:
I have stored 20-50 micron frozen sections of mouse or rat brain in
24/12 well tissue culture plates in PBS in the refrigerator for several
weeks or longer. Immunostaining for tyrosine hydroxylase, dopamine
beta-hydroxylase, GFAP and Mac-1 was not harmed. However, other antigens
may not be so forgiving! The only problem I had is that
mould/fungus/bacteria will eventually start to grow. I would recommend
autoclaving the PBS and changing it every other week, or sooner.
For longer storage you might want to try storing the sections at
minus 20C in a "cryoprotectant" solution of sucrose or ethylene glycol
(and glycerine?), but I have no first hand experience with that.
Geoff
Katy Whalley wrote:
>Hi,
>
>I apologise if this question has been asked before, but I am new to this
>list! I would like to know the best way to store 30 micron sections, cut
>on the cryostat. The sections are PFA-fixed and, after cutting, they are
>in PBS. I want to use them with a free floating immunofluorescence
>protocol, but may need to store some for later use if possible. What is
>the best way to store the sections before staining? Any advice would be
>appreciated!
>
>Katy Whalley, UCL
>
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>
--
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029
mcauliff <@t> umdnj.edu
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