[Histonet] ID rat capillarys in muscle

[Rocan]

Steve,
I would strongly suggest that you look up the papers from Donald M. 
McDonald (UCSF).  Search on medline for author "McDonald DM".  There 
absolutely no other group in the world better for vessel staining in 
vivo.  I do recommend you do it in vivo before euthanasia.   We have 
developed all our  methods his papers.
In brief: you inject the animal with a very small amount (less than 
1ug),  of griffonia isolectin B4 conjugated to a fluorochrome or 
biotin.  Then you must perfuse the animal with 0.1% FRESH 
payaformaldehyde in (PBS).  The perfusion done at 120 mmHg via left 
ventricle; the right atrium is punctured for drainage.  It works like a 
charm!
This procedure will have all the vessels lumens stained with 
fluorescence  and the perfusion prevents vascular collapse.  They have 
modified their techniques so it is important to start with his newest 
papers and go back to older ones as needed for details.   In your case 
it would be great because it would also tell you if the vessels had 
flow.  VEGF can create hemangioma-like networks that are not perfused 
properly .
Contact me directly if you need any details.
Good luck

-----
Dr.Rocio Sierra-Honigmann
Director
Engineered Wound Repair Laboratory
Cedars Sinai Research Institute
310-423-1882
Honigmannr <@t> cshs.org	

On Jun 9, 2005, at 8:14 AM, Steven Coakley wrote:

> Working with ischemic rat hindleg injected with VEGF.  I've read some 
> articles recommending capillary endothelial cell identification with 
> CD 31, Factor VIII and RECA-1,  if anyone has experiance with any of 
> these, others or similar work I'd appreciate the contact.
>
> Steve
>
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