----- Original Message -----From: [hidden email]To: [hidden email]Sent: Monday, January 04, 2010 1:12 PMSubject: Re: Spinning disk requirement ... consider alternative array scan confocalWere you using an EM camera for the spinning disk? I would have thought the EM camera would have better S/N than the PMT's with low signals. Also, how long does it take to capture each frame on the Leica? I had the impression it was 0.5-1 sec/frame which would be much slower than a spinning disk. Dave
On Jan 4, 2010, at 1:40 PM, Carl Boswell wrote:
One system I saw mentioned only casually was the resonant scanner, particularly Leica's. We have a new SP5 dual-head system, and we chose the Leica high-speed system after direct comparisons with DSU and Yokogawa spinning disks. The differences were significant and consistent. As a test specimen, we tracked GFP and DS-Red-labelled vessicle transport and exchange. The Leica provided quite useable images while the spinning disk systems produces unuseable images. The issue was capturing the low intensity signal at a high rate. From my experience, the drawback of spinning disk systems is their inability to do a good job gathering low intensity images. The resonant scanner is noisy, but the 8 kHz speed allows multi-frame averaging. Also the system uses liquid-cooled PMT's. One of the users has tested the system with other specimens and found it to have relatively low photobleaching as compared with the Nikon swept field.Slomething to think about, anyway.cCarl A. Boswell, Ph.D.
Molecular and Cellular Biology
University of Arizona
520-954-7053
FAX 520-621-3709t----- Original Message -----From: [hidden email]To: [hidden email]Sent: Monday, January 04, 2010 10:18 AMSubject: Re: Spinning disk requirement ... consider alternative array scan confocalArray scanners are sold in the US by Nikon and Prairie Technologies. I am not sure if there are other sources and how they compare to the Visitech systems. I have no personal experience with eithe of the systems but I thought the information may help.There are always trade offs with any design, Yokogawa scanheads have been around for years and I understand the design has been improved with the new models. The array scanners are however realtively new and untried.I think you should test the systems and compare by yourself. The best is to get a system to test at your site for an extended time is the ideal situation but difficult to get. I am not convinced that short term demos are the best since many times you may get biased by the skills of the demo crew or you may have problems with a system due to precarious installations or technical problems... the second best is to travel to a location were a system has been in operation for some time, there you also can also talk to experienced users. The problem of traveling to a distant location is that your live specimen preparations may not travel well.Leoncio
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of charu tanwar
Sent: Wednesday, December 30, 2009 11:18 PM
To: [hidden email]
Subject: Re: Spinning disk requirement ... consider alternative array scan confocal
Thank you Dick. I also find it important to have choices in terms of pinhole size.
--- On Wed, 30/12/09, RICHARD BURRY <[hidden email]> wrote:
From: RICHARD BURRY <[hidden email]>
Subject: Re: Spinning disk requirement ... consider alternative array scan confocal
To: [hidden email]
Date: Wednesday, 30 December, 2009, 9:55 PM
Charu
The spining disk is limited to one pinhole size but the 2D array scanning Infinity3 has sever different pinholes. We have the Infinity3 and find the choice of pinholes is very important. For some samples, thinner optical sections (with smaller pinholes) is important and for some samples more intensity (with larger pinholes) is important. Don't forget the camera, I would recommend the most sensitive you can get. Most samples have too few photons! The Hamamatsu backthinned C9100-13 works well for us.
Dick
----- Original Message -----
From: George McNamara <[hidden email]>
Date: Wednesday, December 30, 2009 10:47 am
Subject: Re: Spinning disk requirement ... consider alternative array scan confocal
To: [hidden email]
> Hi Charu,
>
> Check out VT-Infinity and VT-Hawk at Visitech
> http://www.visitech.co.uk/site/products.php The VT-Hawk adds
> FRAP/photoactivation capability to the Infinity. Visitech sold
> Yokogawa's for a long time - Steve Coleman told me in April 2009
> that
> their company was selling off its remaining spinning disk
> inventory
> to focus exclusively on their array scanners. A VT with the
> right
> EMCCD would be a great combination. My biggest concern with
> Visitech
> is how do they support overseas: they are a British company, so
> I am
> referring to both here (USA) and you.
>
> As for Calcium ion flux, the "standard of care" is Fura-2 for
> its
> dynamic range. However, the excitation light path of most
> fluorescence microscopes are such that UV is usually not
> parfocal
> with visible. Until recently, even violet (405 nm) was not
> parfocal
> with green and red (and now only a few expensive objective
> lenses
> are). Parfocality is not critical in widefield Fura-2, but would
> be
> for confocal. The dynamic range of fluorescent protein based
> biosensors is not anywhere near Fura-2, but are getting better,
> for
> example, GCaMP-3, and can be targeted to specific location(s) by
> appropriate targeting sequences.
>
> CARV II - I manage a BD pathway 855 that has a CARV (I?) inside.
> Nice
> to be able to see confocal by eye, but the BD (formerly ATTO)
> software is bad. If you buy a CARV II, be sure to evaluate the
> software that you will be using with it.
>
> Sincerely,
>
> George
>
> At 02:36 AM 12/30/2009, you wrote:
> >Dear All
> >
> >I request all the members to please help me to decide for a
> spinning disk
> >confocal microscope for our facility which is a multiuser
> facility for the
> >whole university. We already have a point scanning confocal
> system from
> >Olympus. We will have another point scanning confocal soon with
> all the live
> >cell imaging accessories (laser based).
> >For very fast phenomenon like measuring Calcium Flux in the
> cells, we want
> >to have a multi point scanning system. But i am unable to
> decide whether i
> >should take a fluorescence based spinning disk or laser based
> spinning disk
> >confocal.
> >Also, please give me some feedback for CARV II (multi point confocal,
> >fluorescence based).
> >Please help.
> >Thanks in advance.
> >
> >Charu Tanwar
> >Imaging Specialist
> >Advanced Instrumentation Research Facility
> >Jawaharlal Nehru University
> >New Delhi
> >India.
>
>
>
>
>
>
>
> George McNamara, Ph.D.
> Image Core Manager
> Analytical Imaging Core Facility
> University of Miami, Miller School of Medicine
> Miami, FL 33136
> [hidden email]
> [hidden email]
> 305-243-8436 office
> http://www.sylvester.org/AICF (Analytical Imaging Core Facility)
> http://www.sylvester.org/AICF/pubspectra.zip (the entire 2000+
> spectra .xlsx file is in the zip file)
> http://home.earthlink.net/~geomcnamara
>
>
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Richard W. Burry, Ph.D.
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