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http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocalDear List,
I am trying to help someone image CFP-labelled mitochondria in mouse brain
slices. Specifically, we are interested in observing the transport of
these organelles along axons.
We have no problem in imaging the mitochondria, the problem is that they
do not move. Excised nerves exhibit rapid bidirectional transport, but in
the brain slices everything is completely static.
Does anyone have experience of imaging anything like this?
We are currently sectioning 300 um slices using a vibratome in oxygenated
neurobasal-A medium and imaging in the same medium less than 30 min after
the brain has been removed. We've tried sectioning in chilled buffer
and warm buffer. We are imaging at 37oC.
Any suggestions gratefully received.
Thanks.
Simon
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