Knocking holes into brain tissue

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From:Peter Bannister <>
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Hello again,

Does anyone have any handy hints for making slices
(60um) of brain tissue permeable to antibodies. We are
currently staining for calcium binding proteins in
different classes of neurones using fluorescent
labels. We also need to maintain the ultrastructure as
much as is possible for subsequent electron microscopy
of synaptic contacts so detergents like triton x100
are out. We have been freeze thawing over liquid
nitrogen 3 times for 30 seconds with excellent
results. However in some cases I feel that we are
getting inadequate penetration of primary antibody.
So, any ideas welcome.

Thanks, Peter Bannister 

Peter Bannister
Senior Research Technician
UCL Med. School
London. UK.

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