Re: Knocking holes into brain tissue

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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.
Dear Peter,
Triton can be used on brain tissue vibratome sections (to improve
penetration of antibodies) and still give a very good morphology (at least
when you can use aldehyde fixation). Hong Yi at Emory  can give you all the
details. You can reach her at
Regards, Peter

Peter van de Plas
Costerweg 5
6702 AA Wageningen
The Netherlands
phone: (31)-317-497676
fax: (31)-317-415955

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