I tend to agree with Dick here, The pinhole array
solutions while less "mature" than the spinning disk confocals work
very well. We have had a Yokagawa head for ages, and the limitation to a
1.4NA objective was significant. About a year ago we bought a Prairie
swept field array scanning confocal. It allows multiple pinholes or slits
sizes (7) depending on your specimen and optics used. It works very
well indeed, and in our hands is more flexible than the Yokogawa head.
Its important to note that the yokogawa head I have is very old and there have
been many advances. However without rapid and simple changing of pinhole
size (as is possible with the Prairie or Visitech solutions) your image will be
compromised when NA is changed.
S.
Simon C. Watkins Ph.D, FRCPath
Professor and Vice Chair, Cell Biology and Physiology
Professor, Immunology
Director, Center for Biologic Imaging
BSTS 225, University of Pittsburgh
3500 Terrace St.
Pittsburgh PA 15261
Tel: 412-352-2277
Fax:412-648-2797
URL: http://www.cbi.pitt.edu
From: Confocal Microscopy
List [mailto:[hidden email]] On Behalf Of RICHARD
BURRY
Sent: Tuesday, February 10, 2009 3:20 PM
To: [hidden email]
Subject: Re: Zeiss SD
Fred
We have a Visitech Infinity3 2D array scanning confocal. It is based on
the same principle as the spinning disk but use an array of pin holes.
This confocal allows seven different sizes of pin holes
with diameter from 10 um to 60 um. Our experience is that is gives
great performance with a Hamamatsu EM13. With the 100x 1.4 N.A. the
optimal pin hole is 30 um but at 60 um the optical section thickness is a
little more than a micron but the intensity is great for even weak fluorescence.
Dick
----- Original Message -----
From: Fred Mast <[hidden email]>
Date: Tuesday, February 10, 2009 2:12 pm
Subject: Re: Zeiss SD
To: [hidden email]
> Does the CSU-X1 allow you to change the disks? I'm still
waiting for a spinning-disk unit that is actually designed for live-cell
imaging. The previous Yokogawa units were optimized for 100x 1,4 NA oil
objectives if I'm not mistaken. Even with relatively thin samples (cultured
cells), you're still introducing spherical aberration... By comparison, has
anyone else looked at the technology being developed in Raphael Yuste's
laboratory? They have combined multiphoton LSM with spatial light modulation
technology to effectively "scan" an entire field at once. I'm not
an expert in this but in my opinion if you could make it fast enough this
seems like a better option to me (they report 60Hz calcium ion imaging). You
can check it out here (under the SLM Microscopy section):
> Fred
D. Mast |
Richard W. Burry, Ph.D.
Department of Neuroscience, College of Medicine
Campus Microscopy and Imaging Facility, Director
The Ohio State University
Associate Editor, Journal of Histochemistry and Cytochemistry
277 Biomedical Research Tower
460 West Twelfth Avenue
Columbus, Ohio 43210
Voice 614.292.2814 Cell 614.638.3345 Fax 614.247.8849
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