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

Renato,
Another suggestion, if you haven't already done this, is to post fix in 2-4% formaldehyde after you have finished staining so your antibodies remain bound and maintain their positions between prep times. This may help.
hank adams

-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Renato Mortara
Sent: Thursday, January 20, 2011 5:04 AM
To: [log in to unmask]
Subject: Is this approach valid ?

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

Hello,

Having being in the first of Jim Pawley's course I feel embarassed to ask
this, but the doubt is haunting me. Here it goes:

The system and approach: nucleoli of eukaryotic cells double immunolabeled
for pairs of antigens. Z-sectioning through the structures (some quite
spherical, others very disperse and granular) the fluorescence intensity
profile of some of these pairs of antigens appear distinct, ie. the axial of
distribution of different antigens varies among samples - that were NOT
necessarily imaged or PREPARED on the same day. 

The question: is it valid to compare these distribution patterns
(intensities along z axis) of antigen pairs between these different samples
(nucleoli of cells A x cells B)?

Many thanks !!

Best

Renato

Renato A. Mortara
Parasitology Division
UNIFESP - Escola Paulista de Medicina
Rua Botucatu, 862, 6th floor
São Paulo, SP
04023-062 
Brazil
Phone: 55 11 5579-8306
Fax:     55 11 5571-1095
email: [log in to unmask]
home page: www.ecb.epm.br/~ramortara