[Histonet] MicroFil

[Barry R Rittman]

Sven
I used Microfil latex several years ago to demonstrate blood vessels in
ginigiva, mostly using dogs for soemone elses project. The following is
from memory as I am at work. 
The Microfil solution is a little viscous but can be perfused into even
the smallest capillaries. We used a peristaltic pump first rinsing out
the blood using saline (with heparin added) until the lips blanched
followed by the final Microfil mixture. All solutions were at 37 degrees
C. The Microfil was prpeared only 15 minutes before use and bubbles
removed using an ultrasonic bath so that they would not become trapped
in small vessels and block them. Care must be taken not to use too much
presure with the pump or else some vesels will be ruptured.
Once the latex had set we then immersed blocks in buffered formalin. 
Some blocks were dehydrated and cleared in methyl salicylate to
visualize the blood vessels. Tissue with bone and teeth were
demineralized using 1% formic acid before dehydrating and clearing.
Other tissue were processed to paraffin wax for evaluation.
The Microfil survived the processing in all procedures. In the paraffin
wax however the Microfil tended to shrink somewhat in diameter so that
the diameter of some vessels appeared initially to be smaller.
We used a combination of blocks and paraffin sections to evaluate blood
vessels in the gingiva.
Never carried out immunofluorescence on these samples but see no logical
reason why this could not be done.
Hope that this helps
Barry


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sven
Terclavers
Sent: Monday, March 15, 2004 4:54 AM
To: Mailinglist_Histology
Subject: [Histonet] MicroFil


Dear listers,

I'm looking for a solution to perfuse bloodvessels which hardens after
cooling down or after a certain timepoint of being liquid to fill not
only larger bloodvessels but also cappillars.  On the internet I found
MicroFil, now I was wondering if some of you already tried this.  I
would like to know the following:

-What is the smallest cappillar that will be filled?
-Can sections be made without the MicroFil falling out of the
cappillars? -Can this MicroFil solution be mixed with e.g. FITC to
perform epi-fluorescence/CLS-microscopy?

Thanks in advance,

Sven Terclavers


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