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Hi dave-

We had white light tirf illuminators on all scopes in the woods hole phys
course for the past 2 yrs and noone used them at all.  The main reason is
NOT ENOUGH LIGHT!  But also, there seems to be out of focus fluorescence
produced by scattered light more too-  I thnk this is what you are seeing.  

Clare M. Waterman-Storer, Ph.D.
Associate Professor
Laboratory for Cell Motility Studies
Department of Cell Biology CB163
The Scripps Research Institute
10550 North Torrey Pines Road
La Jolla CA, 92037 USA
Office:  858-784-9764
Lab:  858-784-9243
Fax:  858-784-9779
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Administartive Assistant: Denise Munoz
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858-784-9964

-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On
Behalf Of David Knecht
Sent: Thursday, November 30, 2006 11:50 AM
To: [log in to unmask]
Subject: white light TIRF

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We have been looking at TIRF systems and recently tested several  
white light TIRF systems. Has anyone directly compared laser vs.  
white light TIRF?  The floating/attached bead demo works fine so it  
looks like TIRF.  However, when we focus up from the coverslip with  
GFP labelled cells, we can see significant fluorescence in focal  
planes that should not be getting excited (5 microns up).  Is this  
white light vs. laser TIRF, or is it that cells present a problem not  
seen with beads.  Dave

Dr. David Knecht
Department of Molecular and Cell Biology
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)