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Hi James,
I'm trying to define the PSF's for our Nikon confocal and came across this
communication of yours from a year ago.  Do you  still have the image data
available online?  The link below doesn't work for me.  Did you finish the
project?
thanks,
Carl


Carl Boswell, Ph.D.
Dept. of Molecular and Cellular Biology
Univ. of Arizona
520-626-8469
520-621-3709 FAX
----- Original Message -----
From: "James Beals" <[log in to unmask]>
To: <[log in to unmask]>
Sent: Thursday, March 06, 2003 8:37 AM
Subject: Re: PSF examples?


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Hi Martin,
I am working on a project similar to what you are looking for.
I am measuring the PSF on our Zeiss 510 Meta.  Using the PSF kit from
Molecular Probes.
The point is to improve our deconvolution processes.
Since I just started, I am still collecting data.
I have posted 2 images on my server.
http://ic3d.med.umich.edu/psfweb/index.htm
The data was collected with a 63x 1.2NA obj. With a 2x digital zoom (0.14um
per pixel)
The subject are 0.175um micro spheres (emitting 505-550nm excite with an
argon laser @ 488nm).
I used the X,Y,Z 1:1 function and collected 26 optical sections. Centering
the sphere.
After many attempts to set the correction collar exactly right (equal ariy
disks north and south)
I settled for less than perfect.  Two sets were taken to compare open
pinhole and 1 airy unit (112um)
The stacks were rendered in Volocity 2.0.
The images show several imperfections.
As far as pinhole alignment, we did that before the test.
I may have some earlier images with misalignment, but I will have to dig
them up.
I hope this helps.
Again, I just started this project and will have more later.
Best




James Beals
Research Associate II
University of Michigan
Department of Cell and Developmental Biology
4779 Medical Science Build. II
[log in to unmask]
lab734.615-8308
fax734.763 -1166

>>> [log in to unmask] 03/05/03 12:15PM >>>
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Dear all confocalists,

I wonder if one can find example pictures of PSF (Point-Spread Functions)
somewhere (measured by CLSM on 90-250nm beads). I would like to see how a
PSF can look like for various individual objectives in reality, including
both the bad and the good ones. Also, I am interested to see how
misalignment in the confocal will/can influence the PSF.

Do someone have experimental PSF examples on the net? Or elsewhere?

Thanks in beforehand,
Martin