CONFOCALMICROSCOPY Archives

March 2003

CONFOCALMICROSCOPY@LISTS.UMN.EDU

Options: Use Monospaced Font
Show Text Part by Default
Show All Mail Headers

Message: [<< First] [< Prev] [Next >] [Last >>]
Topic: [<< First] [< Prev] [Next >] [Last >>]
Author: [<< First] [< Prev] [Next >] [Last >>]

Print Reply
Subject:
From:
James Beals <[log in to unmask]>
Reply To:
Confocal Microscopy List <[log in to unmask]>
Date:
Thu, 6 Mar 2003 10:37:12 -0500
Content-Type:
text/plain
Parts/Attachments:
text/plain (52 lines)
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

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 >>>
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal 

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

ATOM RSS1 RSS2