[Histonet] Immunofluorescence

Natalia Fernandez nunut86 <@t> hotmail.com
Tue Feb 7 09:35:29 CST 2012


Hi everybody!
I have a problem with my experiment.
 
I try to do immunofuorescences in old human brains (parafine sections).
First I thought that the too much colocalisation was due to my antibodies (primary or secondary).
After several tests (Only 1st antibody, only 2nd one, simple staining, inverse simple staining,..) I saw that the tissue itself shows a lot of fluorescence (without antibodies, and even after a treatment against lipofuscine (solution of Potassium permanganate).
 
So my question is: Do you have the same problem?
Do you think that parafine could be the reason of this autofluorescence?!
What can I do?
 
Thank you so much for your answers :)
Natalia 		 	   		  


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