Jim,
Thanks for your answer.
Here is a short description of what we have  done.
Purpose of study: To study the angiogenis process and revascularization of
pancreatic islets transplanted at the renal subcapsular site in the rat. We have
followed the vascular growth and done quantitative analysis of the vascular
density and vessel diameter for a period of couple of weeks. After transplantation
the animal is sutured and left for recovery and transplant uptake. After a period
of 10-12 days the anesthetizes animal was reoperted and the kidney very
carefully exteriorized and positioned in a specifically designed plastic cup
The cup was coupled to an arm which was mounted on specially designed heated
stage with micromanipulator control in x, y,and z directions. It was important toto
to isolate the kidey from body sufficiently so as to avoid image distortions
due to breathing.
FITC labelled dextran was injected in the microcirculation to provide the
contrast enhancement. Argon laser was used for imaging.
 
We have included ascorbic acid in our basthing medium.
We have used Zeiss upright confocal microscope. It has enough vertical
clearence for our purposes.
 
After scanning the transplant we have sutured the animal and reopened it later
for serial observations. We have had problems in this stage. We can do this
operation three successive times- first to do the transplant and then to image
the angiogenesis, but are unable to continue further.
Our main problem at htis stage is-- the rats become tolerant to anesthesia and
do not want to go under. As I increase the dose I end up killing them. Does any
one have a good suggestion about anesthesia?
 
Another problem in 3-d imaging has been the actual transplant site. We have
worked out a technique to keep the islets in packets on the flat side of kidey.
its more of an art.
We have generated our own soft ware for image processing in addition to the
software available from Zeiss.
 
Shanti J. Aggarwal
Research Scientist
Biomedical Engineering Program
ENS 612
University of Texas
Austin, Tx 78712-1084                 512 4717029
Fax 512 4710616
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