I've used lectins in combination with alkaline phosphatase in the past
....you can still get fluorescence if you time your APase staining
appropriately.  You might find the GSI lectin useful, depending on the
tissue you are working with.  It works for vessels in some, but not all,
tissues.  You can find double-labelling protocols for microvessels in
Hansen-Smith, et al,
Microvas.c Res. 36: 199-215, 1989; Hansen-Smith, et al, J. Appl Physiol.
73: 776-80, 1992; Hansen-Smith, et al, Microvasc. Res. 44: 112-16, 1992;
Also, a confocal application for the GSI lectin is described in
Hansen-Smith, et al, Circ. Res. 79: 324-30, 1996.  These reports are for
muscle microvessels, but we have a fair amount of unpublished  info on other
vascular beds. Good luck.
 
Fay Hansen-Smith, Associate Professor
Dept. Biological Sciences
Oakland University
Rochester, MI 48309-4401
810-370-3574
 
 
On Wed, 2 Oct 1996, Michael J. Lyon wrote:
 
> I am looking for some help. I am trying to dissect out some very small
> <50 um blood vessels. The main problem is that I cannot see them once
> the blood has washed out.  I would like to stain the vessels with
> something that is compatible with immunohistochemical procedures using
> confocal microscopy.  I am concerned that the vessel staining will
> interfere with the fluorescence when viewed on the confocal either by
> scattering or autofluorescence.  Since alkaline phosphatase is present
> in most vessels and is still active after paraformaldehyde fixation,
> this should would work for the vessel stain.  I am using rats and most
> likely the method of Mayahara et al., Histochemie 11:88;1967.  Any
> advice would be appreciated.
>
> Thanks
>
> Mike
>