>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 hyi@emory.edu.
Regards, Peter

========================================
Peter van de Plas
AURION
Costerweg 5
6702 AA Wageningen
The Netherlands
phone: (31)-317-497676
fax: (31)-317-415955
http://www.aurion.nl