Microinjection experts; help me out please!

Ary Correa Jr. ac24 at cornell.edu
Sat Feb 25 10:54:41 EST 1995


In article <3il6de$6ud at news.iastate.edu>, letrud at iastate.edu (Louis-Eric
Trudeau) wrote:

> umdoble at mail.cc.umanitoba.ca (Brad Doble) wrote:
> >I am trying to inject cultured neonatal rat cardiac myocytes, and have a
> >problem with the microinjection pipette surging forward when I press the
> >inject button (often leading to ruined pipettes!).  This happens even when
> >the injection pressure (as displayed on the LED readout of the injector)
> >is only 1 psi.  Does anyone know what might be going wrong?   I am using a
> >fluorescent dye to monitor the delivery of the injectant and thus, know
> >that the 1 micrometer diameter tips of the pipettes are not clogged.
> >
> >Any feedback would be greatly appreciated.
> >
> >Thanks in advance,
> >
> >Brad
> >
> >--
> >Brad  Doble                    umdoble at mail.cc.umanitoba.ca
> >                                  or:  bradd at sbrc.umanitoba.ca
> >Department of Physiology, University of Manitoba
> 
> 
> 
> 
> One possibility is that the diameter of the opening of your capillary holder is too large for the O.D. of your pipettes.
> Another possibility is the clamping of your pipette in the holder: is there a suitable O-ring there?
> Good luck.
> 
> Louis-Eric
> Laboratory of cellular signaling
> Iowa State University

My guess is that you are compressing the O-ring in the pipette holder while
injecting.  Try a wider capillar tubing or replace the O-ring for a
thougher one.  If you are controling the pipette movement manually, you can
try to back the pipette wilhe injecting.  Takes some skill but can be done.
 

Good luck

Ary 



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