Dear Colleagues:
 
We are trying to stain unmyelinated calycael dendrites in a slice of
the neuroepithelium of the pigeon vestibular apparatus.  The slices
are 250 microns thick. The slices contain myelinated axons from the
bipolar ganglion and these axons loose their myelin as they penetrate
the basement membrane of the neuroepithelium. The unmyelinated
dendritic axons then terminate as calyceal or bouton endings on hair
cells.  We would like to stain these processes, keep the tissue alive
and patch clamp the processes and hair cells.  We view the tissue
using DIC but I have a confocal.  I followed with interest the
disussion of DIi following the lipid bilayer of axons and dendrites.
Can someone tell me the rate of diffusion or active transport of
DIi and provide me with a first pass recipe.  If you have other dye
alternatives for staining myelinated and unmyelinated neural processes
in tissue that can be kept alive for 6 hours(incubation plus recording
time) I would be extremely grateful to hear about it.
Thanks,
 
Manning Correia
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