[Histonet] HELP!!! - Autofluorescence in the mouse brain - HELP!!!!

[Charles Scouten]

Red Blood cells autofluresce.  Sounds like the brains are not perfused to wash out the red blood cells and ensure rapid penetration of fixative.  You need to perfuse the whole body at sacrifice to get out all red blood cells.  See the following link

http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=471001&catdesc=Histology+Equipment&CatThreeID=674&CatOneID=4&subcatdesc=Sacrifice+Equipment&idsubcategory=21

Cordially,
Charles W.  Scouten, Ph.D. 
myNeuroLab.com 
5918 Evergreen Blvd. 
St. Louis, MO 63134 
Ph: 314 522 0300  
FAX  314 522 0377 
cwscouten <@t> myneurolab.com 
http://www.myneurolab.com 


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Pawel,Pasierbek
Sent: Friday, November 12, 2004 2:19 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] HELP!!! - Autofluorescence in the mouse brain - HELP!!!!

Dear All,

 

I need you help. One of the users of our microscopy facility is doing immunoassaying on mouse brain slices. After PFA fixation he sees autofluorescence from certain regions of the brain. It looks like rather big grains in the cytoplasm. We see it in the FITC and Cy3 channel!
Unfortunately it is also visible in the unfixed preps.

 

I would be very grateful for any hint how to get rid of it. It is important that he still is able to do immunostainigs on the process samples.

 

Thank you very much and have a nice day. 

 

Best regards from Vienna,

Pawel

 

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

Pawel PASIERBEK, PhD

Institute of Molecular Pathology (IMP)

Biooptics Department

Dr. Bohr-Gasse 7

A-1030 Vienna

AUSTRIA

 

Tel. +43 1 797 30 591

Fax +43 1 798 71 53

e-mail: pawel.pasierbek <@t> imp.univie.ac.at <mailto:pawel.pasierbek <@t> imp.univie.ac.at> 

 

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