SCG neurons and DiI staining

Hannah Dvorak DvorakH at starbase1.caltech.edu
Fri Sep 2 13:17:26 EST 1994

I have been staining dissociated cultures of newborn rat superior cervical
ganglion neurons with DiI to try to label the processes.  Staining works
great, in the short term; even the finest processes are lit up.  However,
within a day most of the staining is very diffuse (presumably internalized
to vacuoles) or gone.  More distressingly, however, the neurons tend to
start dying after staining.  Has anyone out there used DiI to stain SCG
neurons?  Any luck?  Any other suggestions for good membrane stains for
labeling fine processes with a fluorescent dye?  (I haven't tried DiO yet,
though we do have some around; we also have a variety of voltage-sensitive
dyes that might be useful.)

Also, I would like to stain individual neurons in cell culture so that I
can see which processes come from particular neurons.  I have tried to do
this by ejecting small drops of DiI solution in DMF (with the surfactant
Pluronic F127 added) directly onto cells using a patch pipette, but the DiI
tends to crystallize and clog up the pipette tip, and in any case the
staining seems to kill the cells, as described above.  Does anyone have any
tips on staining single neurons in vitro?

Hannah Dvorak

Hannah Dvorak
DvorakH at starbase1.caltech.edu
Division of Biology, Caltech, Pasadena CA

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