Zeiss SD
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I was interested to see that Zeiss have gone into partnership with Yokogawa
to launch their own flavour of the spinning disk confocal: http://www.microscopy-analysis.com/news/carl-zeiss-and-yokogawa-launch- spinning-disk-cell-observer-sd-2008 Does this represent an acknowledgment that the 5-live doesn't do everything it says on the tin, or just that it's too expensive? Are there any happy 5-live users out there? |
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I too was intrigued when I saw Zeiss roll out a spinning disk. We're happy with our 5 LIVE. Being able to vary the slit aperture size to match different objectives/samples is important in our multi-user facility. It's also nice to capture two fluorescent channels simultaneously. I wish the detector on the 5 LIVE were EM-CCD but the regular CCD does the job. Images sometimes have vertical line artifacts but those are nicely dealt with by Fourier filtering or dividing by a blank image. Must be lower cost (if that's true) and/or established familiarity of the spinning disk that makes them easy to sell.
-Esteban
On Tue, Feb 10, 2009 at 10:35 AM, Simon Walker <[hidden email]> wrote: I was interested to see that Zeiss have gone into partnership with Yokogawa -- G. Esteban Fernandez, Ph.D. Associate Director Molecular Cytology Core Facility University of Missouri 120 Bond Life Sciences Center Columbia, MO 65211 http://www.biotech.missouri.edu/mcc/ 573-882-4895 573-884-9395 fax |
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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): Cheers, Fred On 10-Feb-09, at 10:55 AM, G. Esteban Fernandez wrote:
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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the Yoko head can be ordered with a choice of disk to match your
primary lens. I was told by Zess the licensing agreement restricts them to selling the Yokogawa head coupled to an inverted stand with a stage enclosure environmental chamber. Regards, Glen On Feb 10, 2009, at 11:10 AM, Fred Mast wrote: > 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): > > http://www.columbia.edu/cu/biology/faculty/yuste/methods.html > > Cheers, > > Fred > > On 10-Feb-09, at 10:55 AM, G. Esteban Fernandez wrote: > >> I too was intrigued when I saw Zeiss roll out a spinning disk. >> We're happy with our 5 LIVE. Being able to vary the slit aperture >> size to match different objectives/samples is important in our >> multi-user facility. It's also nice to capture two fluorescent >> channels simultaneously. I wish the detector on the 5 LIVE were EM- >> CCD but the regular CCD does the job. Images sometimes have >> vertical line artifacts but those are nicely dealt with by Fourier >> filtering or dividing by a blank image. Must be lower cost (if >> that's true) and/or established familiarity of the spinning disk >> that makes them easy to sell. >> >> -Esteban >> >> On Tue, Feb 10, 2009 at 10:35 AM, Simon Walker <[hidden email] >> > wrote: >> I was interested to see that Zeiss have gone into partnership with >> Yokogawa >> to launch their own flavour of the spinning disk confocal: >> http://www.microscopy-analysis.com/news/carl-zeiss-and-yokogawa-launch- >> spinning-disk-cell-observer-sd-2008 >> >> Does this represent an acknowledgment that the 5-live doesn't do >> everything >> it says on the tin, or just that it's too expensive? Are there any >> happy 5-live >> users out there? >> >> >> >> -- >> G. Esteban Fernandez, Ph.D. >> Associate Director >> Molecular Cytology Core Facility >> University of Missouri >> 120 Bond Life Sciences Center >> Columbia, MO 65211 >> >> http://www.biotech.missouri.edu/mcc/ >> >> 573-882-4895 >> 573-884-9395 fax > > 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 > [hidden email] > > > |
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In reply to this post by Fred Mast
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]
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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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 Fred
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Presumably the problem is that the Yokogawa is designed for
x60 NA 1.4 -
it's the magnification as well as the NA that determines
the size of the Airy
disk at the pinhole. So a x20 NA 0.5, which is a
common lens, ought to
work pretty well (one third the mag and ~ one third the
NA).
Guy
Optical Imaging Techniques in Cell Biology From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Watkins, Simon C Sent: Wednesday, 11 February 2009 7:47 AM To: [hidden email] Subject: Re: Zeiss SD 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 Fred
No virus found in this incoming message. No virus found in this outgoing message. |
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Guy
The key is the thickness of the optical section. While a 100x 1.4 N.A. objective with a 30 um pin hole has an optical section thickness of 0.7 um, a 20x 0.45 N.A. objective has an optical section thickness of 6.4 um. So you loose a lot with a 20x not only in optical section thickness but also in through put (because of the lower N.A). Dick 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 ----- Original Message ----- From: Guy Cox <[hidden email]> Date: Wednesday, February 11, 2009 5:44 am Subject: Re: Zeiss SD To: [hidden email]
> No virus found in this outgoing message. |
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In reply to this post by simon walker (BI)
There is also technology called Programmable Array Microscopy which was
developed in the lab of Tom Jovin. Cairn Research Limited was beta testing this about 2 years ago: http://www.cairnweb.com/newsletter/pam.html However I could not find any updates on their new web site: http://www.cairn-research.co.uk/Cairn Perhaps others on the list know where things are with PAM. Basically it is a programmable array so you can open as many local array elements to essentially vary the size of the pinhole. You can also simulate any pattern you like so you could make it perform like swept field or like a spinning disk and so on. One of the points I understood is you can also collect the out of focus light for use in deconvolution unlike other systems. I'm not sure how this is done though. It sounds like there is a lot of promise but as I said I'm not sure where things are now. There are also primary research articles out of the Jovin lab. Sincerely, Claire |
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In reply to this post by RICHARD BURRY
Sure, but that applies to any confocal - optical section
thickness
goes as the inverse square of the NA. By and large if
you want
optimal optical sectioning you won't use a spinning disk
anyway.
My point was that there will almost always be more than one
lens
which matches a given pinhole
size.
Guy
Optical Imaging Techniques in Cell Biology From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of RICHARD BURRY Sent: Thursday, 12 February 2009 12:20 AM To: [hidden email] Subject: Re: Zeiss SD The key is the thickness of the optical section. While a 100x 1.4 N.A. objective with a 30 um pin hole has an optical section thickness of 0.7 um, a 20x 0.45 N.A. objective has an optical section thickness of 6.4 um. So you loose a lot with a 20x not only in optical section thickness but also in through put (because of the lower N.A). Dick 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 ----- Original Message ----- From: Guy Cox <[hidden email]> Date: Wednesday, February 11, 2009 5:44 am Subject: Re: Zeiss SD To: [hidden email]
>
No virus found in this outgoing message. No virus found in this incoming message. No virus found in this outgoing message. |
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