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The difference will come with the SA correction.  TIRF
lenses (at least the Leica one) actually have values for
different temperatures on the correction ring.  This is 
working with oil, which changes RI rather a lot with 
temperature.  In water the difference will be smaller, but
it's still worth setting the correction collar visually
rather than by trusting the scale.  Once you have set it
correctly there should be no real difference in PSF.

                                              Guy

Optical Imaging Techniques in Cell Biology
by Guy Cox    CRC Press / Taylor & Francis
    http://www.guycox.com/optical.htm
______________________________________________
Associate Professor Guy Cox, MA, DPhil(Oxon)
Electron Microscope Unit, Madsen Building F09,
University of Sydney, NSW 2006
______________________________________________
Phone +61 2 9351 3176     Fax +61 2 9351 7682
Mobile 0413 281 861
______________________________________________
     http://www.guycox.net
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Carl Boswell
Sent: Friday, 8 February 2008 5:58 AM
To: [log in to unmask]
Subject: heat and PSF

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Hi all,
Does anyone have experience with characterizing PSF's (or other aspects of the optics) at different temperatures?  For instance, how much effect  is there on an objective when the temp is raised from room temp (25C) to physiologically relevant 37C?
Thanks,
Carl

Carl A. Boswell, Ph.D.
Molecular and Cellular Biology
University of Arizona
520-954-7053
FAX 520-621-3709 

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