Planaria

aloisia schmid a-schmi at ux1.cso.uiuc.edu
Tue Mar 12 20:08:49 EST 1996


In article <rhoskins-0503961604060001 at mac55.zoology.uwa.edu.au>,
rhoskins at uniwa.uwa.edu.au 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 ux1.cso.uiuc.edu



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