Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Marianne,
It is likely that the autofluorescence that you are getting in the pancreas is from lipofuschin deposits, particularly seen in slightly older animals or tissues. This can be quenched using Sudan Black staining or copper sulphate treatment, thereby enabling you to use the green fluorescence channel. We have previously used the Sudan Black treament in retinal wholemounts and sections and in cardiac tissue with excellent results. There is some basic methodological information (and references to the original publications) on our website:

http://biaf.uwa.edu.au/biaf/autofluor.htm

I hope this helps. Good luck.
Paul Rigby

---------------

Dr Paul J Rigby
Director and Senior Lecturer
Biomedical Imaging & Analysis Facility (M510)
The University of Western Australia
35 Stirling Highway, Crawley, WA, 6009
Australia
Ph (61-8) 9346 2819
Fx (61-8) 9346 3469


-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]]On
Behalf Of Marianne Martinic
Sent: Thursday, 19 May 2005 12:36 AM
To: [log in to unmask]
Subject: Tracking of live cells


Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hello everybody,

I am just starting using the confocal microscopy and would like to ask
if anyone of you already traced activated and labeled CD4 and CD8 T
cells in the pancreas? Due to the high autofluorescence of the
pancreas, I thought of chosing PHK26 from Sigma together with DiD from
Invitrogen to label the T cells in order to avoid the green
fluorescence channel.

Thank you very much for any advice,

Marianne
--
Marianne Martinic, Ph.D.
La Jolla Institute for Allergy and Immunology
Division of Developmental Immunology DI-3
10355 Science Center Drive
San Diego, CA 92121
U.S.A.
Phone: (858) 678 4581
Fax: (858) 558 3579
[log in to unmask]