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Hello all,

I would like some input into an idea I have for obtaining a PSF. I  
study peroxisomes using yeast as a model system for understanding how  
they are created. In yeast, peroxisomes have a well characterized  
morphology, being spherical organelles with a diameter of 100 to  
200nm. Other than this size variability I think they are excellent  
candidates for obtaining a PSF as they can be easily, fluorescently  
labelled (by targeting fluorescent protein chimeras to their matrix),  
are similar in size to what is typically used to obtain PSF's, and are  
"embedded" in the sample. I do a lot of live cell imaging, using a  
LSM510 Meta and am always looking for ways to improve my system. As a  
result most of my images are fairly noisy and I rely on deconvolution  
to remove the noise, and improve contrast and resolution. My initial  
attempts at using peroxisomes for this purpose have provided me with a  
PSF that is slightly different from what I obtain with fluorescent  
beads (the peroxisome derived PSF is less symmetrical) and provides,  
in my estimation, a more realistic result. Your thoughts and concerns  
on this idea would be most welcome.

Fred

Fred D. Mast
Department of Cell Biology
Medical Sciences Building Room 5-14
University of Alberta
Edmonton, Alberta, T6G 2H7
Canada

Tel: 1-780-492-7407
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