aloisia schmid a-schmi at
Tue Mar 12 20:08:49 EST 1996

In article <rhoskins-0503961604060001 at>,
rhoskins at wrote:

> I need to know of a tracer to trace planaria nervous system. I used Di I
> dissolved in DMSO and directly injected to the brain, The planaria
> disintergrated probably due to the DMSO. I wonder if there is a water
> soluable tracer to use which will travel up the axons.

Try dissolving the DiI in Wesson Vegetable Oil (but any oil will
do....even the ones without 'wessonality').  I use 1 mg/ml and usually
make about 5 or 6 mls at a time.  After weighing out the dry DiI and
putting it into an eppendorf containing 1 ml of vegetable oil, I vortex
the heck out of it for about 10'.  Just sit there with the tubes on a
vortex and hold it down.  Very annoying feeling in your hands.  Then you
let it sit in a beaker of ice for about a half an hour and then sonicate
it;  I generally do five second pulses at maximum out put, with five
minute rest periods in between.  I do that 12 times (total of one minute
of sonication.)  Then spin down in a microfuge for about 10' at 13,000
RPM.  Then aliquot the stuff on top.  You can store this in the -20 for up
to a year, and even though this is time consuming, you generally get nice
results.  Oh, I always spin an aliquot before using it after thawing.  I
don't know how you are actually injecting this, but I have labeled unfixed
and fixed cells in culture, and drosophila neuroblasts with no problem and
those are pretty aqueous milieus to be injecting a drop into, so this
should work for you too.  The thing is I think you have to be able to
visualize where you want the drop to go.

If you have any other questions, let me know!  Good Luck!

Alice Schmid

Aloisia Schmid
Howard Huges Medical Institute/Morrill Hall
University of Illinois
505 S. Goodwin Ave, 618 Morrill Hall
Urbana, IL 61801
Email: A-schmi at

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