live adipose tissue

[leoncio vergara]

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We are working on a project involving FLIM imaging of live adipose tissue. We
have experience imaging and keeping alive other tissues but this is the first
time we will attempt to work with fat tissue.
The project involves receiving fresh tissue from biopsies at the begginning of
the day and then keeping them alive during several hrs while we complete the
FLIM imaging sessions on a multiphoton microscope.
The question is how to keep the samples viable outside of the microscope and
on the microscope. We do have the type of equipment commonly used to keep
brain slices alive (imaging chambers, stage incubator, perfusion devices),
however so far I have not found a description about the optimal requirements
to keep adipose tissue viable so we can perform studies that would have
physiological significance.
Also, is there any parameter or dye we can use to monitor viability?

Thanks in advance for your help.

Leoncio Vergara

[Boswell, Carl A - (cboswell)]

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Hi Leoncio,

My guess would be that there are no particular conditions that would be
preferred for adipose vs. other mammalian tissues.  A standard culture
medium with good oxygenation and temperature control should work fine.  The
biggest issues are oxygen diffusion into thick tissue and exchange of waste
and nutrients, so the thinner the layer of fat, the better.

As for monitoring viability, Molecular Probes has a Live/Dead staining mix,
but again diffusion into tissue may be an issue, as well as possibly
confounding signals with whatever you are looking at with FLIM.  Maybe
applying the assessment after the FLIM measurement would give you an
indicaiton of overall tissue health.

Good luck,

c

Carl A. Boswell, Ph.D.
Molecular and Cellular Biology
University of Arizona
520-954-7053
FAX [phone]
----- Original Message -----
From: "Leoncio Vergara" <[hidden email]>
To: <[hidden email]>
Sent: Monday, September 20, 2010 12:38 PM
Subject: live adipose tissue


============================================================
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
============================================================

We are working on a project involving FLIM imaging of live adipose tissue.
We
have experience imaging and keeping alive other tissues but this is the
first
time we will attempt to work with fat tissue.
The project involves receiving fresh tissue from biopsies at the begginning
of
the day and then keeping them alive during several hrs while we complete the
FLIM imaging sessions on a multiphoton microscope.
The question is how to keep the samples viable outside of the microscope and
on the microscope. We do have the type of equipment commonly used to keep
brain slices alive (imaging chambers, stage incubator, perfusion devices),
however so far I have not found a description about the optimal requirements
to keep adipose tissue viable so we can perform studies that would have
physiological significance.
Also, is there any parameter or dye we can use to monitor viability?

Thanks in advance for your help.

Leoncio Vergara