spinning disk confocal Vs deltavision?

[zhan cheng]

Hello everyone,
   Our facility is planning to buy a live cell imaging setup, spinning disk
confocal and deltavision seem both good. We are concering about imaging
quality, imaging speed, phototoxin, and photo bleaching for long term imaging.
Could you share your experience? some time lapse our imaging maybe last over
12 hours, so  sometimes the data is huge. I know the  deltavision system's
imaging quality is good after deconvolution, but I worry about the 3D
deconvolution's speed, especially for huge data.
    Thanks.
    zhan cheng

[Alberto Diaspro]

Spinning in case

Inviato da iPhone

Il giorno 23/feb/2010, alle ore 02.50, zhan cheng  
<[hidden email]> ha scritto:

> Hello everyone,
>   Our facility is planning to buy a live cell imaging setup,  
> spinning disk
> confocal and deltavision seem both good. We are concering about  
> imaging
> quality, imaging speed, phototoxin, and photo bleaching for long  
> term imaging.
> Could you share your experience? some time lapse our imaging maybe  
> last over
> 12 hours, so  sometimes the data is huge. I know the  deltavision  
> system's
> imaging quality is good after deconvolution, but I worry about the 3D
> deconvolution's speed, especially for huge data.
>    Thanks.
>    zhan cheng

[Hu Xian-3]

Dear Zhan Cheng,

We have one DeltaVision(one year) and one PerkinElmer Vox(owned two
month). Although they are both well know for live-cell imaging, the
reasons why they are suitable for live-cell imaging aren't quite the
same. Let me try to answer your question individually first,

Image Quality: Both systems are camera based. Default camera for DV is
HQ2, 512X512, cooled but not EMCCD; you can choose to add a second
EMCCD, cascadeII 512X512, but only this one as I know and I didn't have
a chance to see it in action; microscope for DV is olympus IX71 series.
Spinning disk, you can choose what camera you want to have, we have two
Hamamatsu EMCCD on ours; you can choose whatever microscope body you
want as well.

Imaging speed: Speed of spinning disk is limited by camera, and I think
the speed limiting step for DV is the shutter. Also, we have piezo Z for
our spinning disk, and not sure whether we can have that on the DV. So
it seems that our spinning disk can be much faster than our DV.

Phototoxicity/Photobleaching/Long term imaging: They should both quite
good for 12 hours imaging, if used properly. But the out come for this
competition heavily depends on the experiment setting.

Deconvolution speed: Ask DV to sent you a faster computer. They have
batch processor, you can run your decon when you are snoozing. (beware
that decon can be quite tricky, your results can be beautiful but
wrong). PE spinning disk has decon/deblur too.

And if you are purchasing for a core facility, these might be your
concern as well: a. price(well...), b. Most of my new user found it
rather easy to learn and handle DV, but they always have to come back
for second or third training for spinning disk. But more experienced
user loves to play with spinning disk. c. maintenance(cost, service
availability,service quality)..

Hope that helps.

Thanks.

Regards,
Edna




zhan cheng wrote:

> Hello everyone,
>    Our facility is planning to buy a live cell imaging setup, spinning disk
> confocal and deltavision seem both good. We are concering about imaging
> quality, imaging speed, phototoxin, and photo bleaching for long term imaging.
> Could you share your experience? some time lapse our imaging maybe last over
> 12 hours, so  sometimes the data is huge. I know the  deltavision system's
> imaging quality is good after deconvolution, but I worry about the 3D
> deconvolution's speed, especially for huge data.
>     Thanks.
>     zhan cheng
>
>  

[Boswell, Carl A - (cboswell)]

Two points to make.

First, while my only experience with spinning disks have been multiple demos
while looking for high speed imaging, the sensitivity issue seemed
significant.  The SD systems we had to look at struggled mightily when the
samples were dim, while the DV could extract meaningful data from the same
thing.  Second, the pixel size of EMCCD's are going to effect resolution
compared with the CoolSnap HQ (1024x1024).

The photobleaching/toxicity should be similar, so the bonus I've seen from
SD is the speed.  In our hands, the limiting factor for imaging speed with
the DV was the exposure time, again talking about dim samples.  Shutter time
was a trivial issue.

From my view, your choices are resolution and sensitivity vs. speed.

Good luck,
c

Carl A. Boswell, Ph.D.
Molecular and Cellular Biology
University of Arizona
520-954-7053
FAX [phone]
----- Original Message -----
From: "zhan cheng" <[hidden email]>
To: <[hidden email]>
Sent: Monday, February 22, 2010 6:50 PM
Subject: spinning disk confocal Vs deltavision?


Hello everyone,
   Our facility is planning to buy a live cell imaging setup, spinning disk
confocal and deltavision seem both good. We are concering about imaging
quality, imaging speed, phototoxin, and photo bleaching for long term
imaging.
Could you share your experience? some time lapse our imaging maybe last over
12 hours, so  sometimes the data is huge. I know the  deltavision system's
imaging quality is good after deconvolution, but I worry about the 3D
deconvolution's speed, especially for huge data.
    Thanks.
    zhan cheng

[Steve Bagley]

Hi

We have both a deltavision (seven years with several upgrades) and a roper csu22 spinning disk system (one year). Both have the same camera (EMCCD cascade), filters (ET-Chroma) and objective lenses

In our hands we find that

i) when live cell imaging fibre like structures (cytoskeletal elements) the deltavision system seems to present the better results, when imaging spots (e.g. spindle pole bodies) the spinning disk system seems more suited

 

ii) the limiting factor on the spinning disk is exposure time. When going below 50msec the image quality drops due to the rotational speed of the disk. For very fast imaging the deltavision presents more favourable capture times. I believ that with the x1 you can go a little faster in capture speed without deterimentally effecting image quality but the scanner rotational speed is around the same.

 

iii) overall we have seen slightly less photo-toxcity on our spinning disk system. This maybe due to the model of Olympus microscope that our systems are on (DV olympus IX71 SD olympus IX81).

iv) on both systems we have full environmental chambers, bioptechs chambers, objective heaters and the cellasic microfluidic system. Both systems are well suited to enviromental control.

v) I guess the draw back is the range of wavelengths available to the spinning disk via the lasers. We have the full 'sedat filter' range 406/491/555/643 whereas the DV is not limited by wavelength selection.

vi) with both systems the data is deconvolved afterwards, the SD to further vastly improve the axial resolution. All data goes through the same 3D imaging and analysis software. For deconvolution we have scaled up with multiprocessors and linux with Huygens, both data streams (DV and SD) can be processed this way with the same software.

vii) we have been considering the Princeton ProEM 1024B to increase sensitivity and photoefficiency, however the minimum usable capture rate of the spinning disk 50msec still could not be exceeded without seriously effecting image quality. The advantage would be a reduction in the amount of laser light used thus a reduction in the rate of photo-toxicity.

I hope this has helped, all the best

 

steve

Steve Bagley

Head of Imaging

Imaging Facility

Cancer Research UK

Paterson Institute for Cancer Research

University of Manchester

Wilmslow Road

Manchester

M20 9BX

UK

www.paterson.man.ac.uk

 

 

----- Original Message -----

From: "zhan cheng" <[hidden email]>

To: <[hidden email]>

Sent: Monday, February 22, 2010 6:50 PM

Subject: spinning disk confocal Vs deltavision?

Hello everyone,

   Our facility is planning to buy a live cell imaging setup, spinning disk

confocal and deltavision seem both good. We are concering about imaging

quality, imaging speed, phototoxin, and photo bleaching for long term

imaging.

Could you share your experience? some time lapse our imaging maybe last over

12 hours, so  sometimes the data is huge. I know the  deltavision system's

imaging quality is good after deconvolution, but I worry about the 3D

deconvolution's speed, especially for huge data.

    Thanks.

    zhan cheng

---

This email is confidential and intended solely for the use of the person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Paterson Institute for Cancer Research or the University of Manchester. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible.

[Vitaly Boyko]

Hi Steve,

 

It is all true except for the "the draw back is the range of wavelengths available to the spinning disk via the lasers"

 

375mm,

405 nm,

440nm,

488nm,

505nm,

515nm,

532nm,

561nm,

568nm,

593nm,

633nm,

656nm ...

 

Finally, several companies have become customer-friendly (Zeiss, Nikon and others). For example, 375-440-505-568-656 nm combination sounds very attractive (a "downgrade" to three-four lines may look more practical). If someone would need a custom dichroic, talk to Chroma (big names can also talk to Semrock, for the bulk orders only)

Good luck,

 

Vitaly

 

---

From: Steve Bagley <[hidden email]>
To: [hidden email]
Sent: Thu, February 25, 2010 11:19:21 AM
Subject: Re: spinning disk confocal Vs deltavision?

Hi

We have both a deltavision (seven years with several upgrades) and a roper csu22 spinning disk system (one year). Both have the same camera (EMCCD cascade), filters (ET-Chroma) and objective lenses

In our hands we find that

i) when live cell imaging fibre like structures (cytoskeletal elements) the deltavision system seems to present the better results, when imaging spots (e.g. spindle pole bodies) the spinning disk system seems more suited

 

ii) the limiting factor on the spinning disk is exposure time. When going below 50msec the image quality drops due to the rotational speed of the disk. For very fast imaging the deltavision presents more favourable capture times. I believ that with the x1 you can go a little faster in capture speed without deterimentally effecting image quality but the scanner rotational speed is around the same.

 

iii) overall we have seen slightly less photo-toxcity on our spinning disk system. This maybe due to the model of Olympus microscope that our systems are on (DV olympus IX71 SD olympus IX81).

iv) on both systems we have full environmental chambers, bioptechs chambers, objective heaters and the cellasic microfluidic system. Both systems are well suited to enviromental control.

v) I guess the draw back is the range of wavelengths available to the spinning disk via the lasers. We have the full 'sedat filter' range 406/491/555/643 whereas the DV is not limited by wavelength selection.

vi) with both systems the data is deconvolved afterwards, the SD to further vastly improve the axial resolution. All data goes through the same 3D imaging and analysis software. For deconvolution we have scaled up with multiprocessors and linux with Huygens, both data streams (DV and SD) can be processed this way with the same software.

vii) we have been considering the Princeton ProEM 1024B to increase sensitivity and photoefficiency, however the minimum usable capture rate of the spinning disk 50msec still could not be exceeded without seriously effecting image quality. The advantage would be a reduction in the amount of laser light used thus a reduction in the rate of photo-toxicity.

I hope this has helped, all the best

 

steve

Steve Bagley

Head of Imaging

Imaging Facility

Cancer Research UK

Paterson Institute for Cancer Research

University of Manchester

Wilmslow Road

Manchester

M20 9BX

UK

www.paterson.man.ac.uk

 

 

----- Original Message -----

From: "zhan cheng" <[hidden email]>

To: <[hidden email]>

Sent: Monday, February 22, 2010 6:50 PM

Subject: spinning disk confocal Vs deltavision?

Hello everyone,

   Our facility is planning to buy a live cell imaging setup, spinning disk

confocal and deltavision seem both good. We are concering about imaging

quality, imaging speed, phototoxin, and photo bleaching for long term

imaging.

Could you share your experience? some time lapse our imaging maybe last over

12 hours, so  sometimes the data is huge. I know the  deltavision system's

imaging quality is good after deconvolution, but I worry about the 3D

deconvolution's speed, especially for huge data.

    Thanks.

    zhan cheng

---

This email is confidential and intended solely for the use of the person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Paterson Institute for Cancer Research or the University of Manchester. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible.

[Steve Baxter-2]

Hi Steve,

First a short disclaimer - I work for PerkinElmer!

With the spinning disk systems you can start to get scanning artefacts at short exposure times, this is caused by partial disk segments being exposed. You can get around this by increasing the exposure time (which greatly reduces the effect of the partial sectors but also reduces the speed of the system) or by carefully synchronising the camera exposure to the disk speed. We do the latter with the UltraVIEW ERS and VoX, we find that we can get exposures in the 1 ms range with minimal scan-line artefacts.

Cheers,

Steve

On 25 Feb 2010, at 16:19, Steve Bagley wrote:

>
> Hi
>
> We have both a deltavision (seven years with several upgrades) and a roper csu22 spinning disk system (one year). Both have the same camera (EMCCD cascade), filters (ET-Chroma) and objective lenses
>
>
> In our hands we find that
>
>
> i) when live cell imaging fibre like structures (cytoskeletal elements) the deltavision system seems to present the better results, when imaging spots (e.g. spindle pole bodies) the spinning disk system seems more suited
>
>  
>
> ii) the limiting factor on the spinning disk is exposure time. When going below 50msec the image quality drops due to the rotational speed of the disk. For very fast imaging the deltavision presents more favourable capture times. I believ that with the x1 you can go a little faster in capture speed without deterimentally effecting image quality but the scanner rotational speed is around the same.
>
>  
> iii) overall we have seen slightly less photo-toxcity on our spinning disk system. This maybe due to the model of Olympus microscope that our systems are on (DV olympus IX71 SD olympus IX81).
>
> iv) on both systems we have full environmental chambers, bioptechs chambers, objective heaters and the cellasic microfluidic system. Both systems are well suited to enviromental control.
>
>
> v) I guess the draw back is the range of wavelengths available to the spinning disk via the lasers. We have the full 'sedat filter' range 406/491/555/643 whereas the DV is not limited by wavelength selection.
>
> vi) with both systems the data is deconvolved afterwards, the SD to further vastly improve the axial resolution. All data goes through the same 3D imaging and analysis software. For deconvolution we have scaled up with multiprocessors and linux with Huygens, both data streams (DV and SD) can be processed this way with the same software.
>
>
> vii) we have been considering the Princeton ProEM 1024B to increase sensitivity and photoefficiency, however the minimum usable capture rate of the spinning disk 50msec still could not be exceeded without seriously effecting image quality. The advantage would be a reduction in the amount of laser light used thus a reduction in the rate of photo-toxicity.
>
>
> I hope this has helped, all the best
>
>  
> steve
>
>
>
> Steve Bagley
>
> Head of Imaging
>
> Imaging Facility
>
> Cancer Research UK
>
> Paterson Institute for Cancer Research
>
> University of Manchester
>
> Wilmslow Road
>
> Manchester
>
> M20 9BX
>
> UK
>
> www.paterson.man.ac.uk
>
>
>  
>  
> ----- Original Message -----
>
> From: "zhan cheng" <[hidden email]>
>
> To: <[hidden email]>
>
> Sent: Monday, February 22, 2010 6:50 PM
>
> Subject: spinning disk confocal Vs deltavision?
>
>
> Hello everyone,
>
>    Our facility is planning to buy a live cell imaging setup, spinning disk
>
> confocal and deltavision seem both good. We are concering about imaging
>
> quality, imaging speed, phototoxin, and photo bleaching for long term
>
> imaging.
>
> Could you share your experience? some time lapse our imaging maybe last over
>
> 12 hours, so  sometimes the data is huge. I know the  deltavision system's
>
> imaging quality is good after deconvolution, but I worry about the 3D
>
> deconvolution's speed, especially for huge data.
>
>     Thanks.
>
>     zhan cheng
>
> This email is confidential and intended solely for the use of the person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Paterson Institute for Cancer Research or the University of Manchester. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible.

Steve Baxter
R&D CoE Leader
PerkinElmer Coventry
 
Email : [hidden email]  
Telephone: [phone]
Mobile/Cell: [phone]
Fax: [phone]

[Andrea Latini-2]

Dear Dr. Zhan Cheng and dear List,

I think you can solve all problems by using a faster spinning disk confocal
system, i.e. the CARV-II.
this is a 7500 rpm spinning disk unit and I've been able to test it either
with a low resolution-high sensitivity Photometrics EVOLVE 512x512 and a
less sensitive- higher resolution CoolSnap EZ.
Results:
with the Evolve EMCCD I've been down to 5ms exposure time without any loss
due to slow spinning disk speed, i.e. the CSU-22 system (this spins at less
than 1800rpm).

with the CoolSnap camera system I've been able to collect very good images
(i.e. S/N > 5), even at 19ms exposure time.

Of course, due to the CARV-II very high rotation speed, the only limitation
is from the CCD system speed.

I'm ready to provide you with all pieces of information you may need.

Regards.

Andrea Latini, PhD
Crisel Instruments Srl, ITALY
[hidden email]

On Thu, 25 Feb 2010 16:19:21 -0000, Steve Bagley <[hidden email]> wrote:

>
>Hi
>
>We have both a deltavision (seven years with several upgrades) and a
>roper csu22 spinning disk system (one year). Both have the same camera
>(EMCCD cascade), filters (ET-Chroma) and objective lenses
>
>In our hands we find that
>
>i) when live cell imaging fibre like structures (cytoskeletal elements)
>the deltavision system seems to present the better results, when imaging
>spots (e.g. spindle pole bodies) the spinning disk system seems more
>suited
>
>ii) the limiting factor on the spinning disk is exposure time. When
>going below 50msec the image quality drops due to the rotational speed
>of the disk. For very fast imaging the deltavision presents more
>favourable capture times. I believ that with the x1 you can go a little
>faster in capture speed without deterimentally effecting image quality
>but the scanner rotational speed is around the same.
>
>iii) overall we have seen slightly less photo-toxcity on our spinning
>disk system. This maybe due to the model of Olympus microscope that our
>systems are on (DV olympus IX71 SD olympus IX81).
>
>iv) on both systems we have full environmental chambers, bioptechs
>chambers, objective heaters and the cellasic microfluidic system. Both
>systems are well suited to enviromental control.
>
>v) I guess the draw back is the range of wavelengths available to the
>spinning disk via the lasers. We have the full 'sedat filter' range
>406/491/555/643 whereas the DV is not limited by wavelength selection.
>
>vi) with both systems the data is deconvolved afterwards, the SD to
>further vastly improve the axial resolution. All data goes through the
>same 3D imaging and analysis software. For deconvolution we have scaled
>up with multiprocessors and linux with Huygens, both data streams (DV
>and SD) can be processed this way with the same software.
>
>vii) we have been considering the Princeton ProEM 1024B to increase
>sensitivity and photoefficiency, however the minimum usable capture rate
>of the spinning disk 50msec still could not be exceeded without
>seriously effecting image quality. The advantage would be a reduction in
>the amount of laser light used thus a reduction in the rate of
>photo-toxicity.
>
>I hope this has helped, all the best
>
>steve
>
>
>Steve Bagley
>Head of Imaging
>Imaging Facility
>Cancer Research UK
>Paterson Institute for Cancer Research
>University of Manchester
>Wilmslow Road
>Manchester
>M20 9BX
>UK
>www.paterson.man.ac.uk
>
>
>
>
>
>
>----- Original Message -----
>From: "zhan cheng" <[hidden email]>
>To: <[hidden email]>
>Sent: Monday, February 22, 2010 6:50 PM
>Subject: spinning disk confocal Vs deltavision?
>
>
>Hello everyone,
>   Our facility is planning to buy a live cell imaging setup, spinning
>disk
>confocal and deltavision seem both good. We are concering about imaging
>quality, imaging speed, phototoxin, and photo bleaching for long term
>imaging.
>Could you share your experience? some time lapse our imaging maybe last
>over
>12 hours, so  sometimes the data is huge. I know the  deltavision
>system's
>imaging quality is good after deconvolution, but I worry about the 3D
>deconvolution's speed, especially for huge data.
>    Thanks.
>    zhan cheng
>--------------------------------------------------------
>This email is confidential and intended solely for the use of the person(s)

('the intended recipient') to whom it was addressed. Any views or opinions
presented are solely those of the author and do not necessarily represent
those of the Paterson Institute for Cancer Research or the University of
Manchester. It may contain information that is privileged & confidential
within the meaning of applicable law. Accordingly any dissemination,
distribution, copying, or other use of this message, or any of its contents,
by any person other than the intended recipient may constitute a breach of
civil or criminal law and is strictly prohibited. If you are NOT the
intended recipient please contact the sender and dispose of this e-mail as
soon as possible.  
>

[George Peeters-2]

Actually the CSU-22 spins at 5000 rpms and csux up to 10,000 rpm or  
2000 scans / sec (12 scans per rotation).
1800 rpm (360 scans / sec) is the default speed that can be adjusted  
from software to the highest speed.

George Peeters
Solamere Technology Group

Sent from my iPhone

On Mar 1, 2010, at 12:25 PM, Andrea Latini <ricco@CRISEL-
INSTRUMENTS.IT> wrote:

> Dear Dr. Zhan Cheng and dear List,
>
> I think you can solve all problems by using a faster spinning disk  
> confocal
> system, i.e. the CARV-II.
> this is a 7500 rpm spinning disk unit and I've been able to test it  
> either
> with a low resolution-high sensitivity Photometrics EVOLVE 512x512  
> and a
> less sensitive- higher resolution CoolSnap EZ.
> Results:
> with the Evolve EMCCD I've been down to 5ms exposure time without  
> any loss
> due to slow spinning disk speed, i.e. the CSU-22 system (this spins  
> at less
> than 1800rpm).
>
> with the CoolSnap camera system I've been able to collect very good  
> images
> (i.e. S/N > 5), even at 19ms exposure time.
>
> Of course, due to the CARV-II very high rotation speed, the only  
> limitation
> is from the CCD system speed.
>
> I'm ready to provide you with all pieces of information you may need.
>
> Regards.
>
> Andrea Latini, PhD
> Crisel Instruments Srl, ITALY
> [hidden email]
>
> On Thu, 25 Feb 2010 16:19:21 -0000, Steve Bagley <[hidden email]
> > wrote:
>
>>
>> Hi
>>
>> We have both a deltavision (seven years with several upgrades) and a
>> roper csu22 spinning disk system (one year). Both have the same  
>> camera
>> (EMCCD cascade), filters (ET-Chroma) and objective lenses
>>
>> In our hands we find that
>>
>> i) when live cell imaging fibre like structures (cytoskeletal  
>> elements)
>> the deltavision system seems to present the better results, when  
>> imaging
>> spots (e.g. spindle pole bodies) the spinning disk system seems more
>> suited
>>
>> ii) the limiting factor on the spinning disk is exposure time. When
>> going below 50msec the image quality drops due to the rotational  
>> speed
>> of the disk. For very fast imaging the deltavision presents more
>> favourable capture times. I believ that with the x1 you can go a  
>> little
>> faster in capture speed without deterimentally effecting image  
>> quality
>> but the scanner rotational speed is around the same.
>>
>> iii) overall we have seen slightly less photo-toxcity on our spinning
>> disk system. This maybe due to the model of Olympus microscope that  
>> our
>> systems are on (DV olympus IX71 SD olympus IX81).
>>
>> iv) on both systems we have full environmental chambers, bioptechs
>> chambers, objective heaters and the cellasic microfluidic system.  
>> Both
>> systems are well suited to enviromental control.
>>
>> v) I guess the draw back is the range of wavelengths available to the
>> spinning disk via the lasers. We have the full 'sedat filter' range
>> 406/491/555/643 whereas the DV is not limited by wavelength  
>> selection.
>>
>> vi) with both systems the data is deconvolved afterwards, the SD to
>> further vastly improve the axial resolution. All data goes through  
>> the
>> same 3D imaging and analysis software. For deconvolution we have  
>> scaled
>> up with multiprocessors and linux with Huygens, both data streams (DV
>> and SD) can be processed this way with the same software.
>>
>> vii) we have been considering the Princeton ProEM 1024B to increase
>> sensitivity and photoefficiency, however the minimum usable capture  
>> rate
>> of the spinning disk 50msec still could not be exceeded without
>> seriously effecting image quality. The advantage would be a  
>> reduction in
>> the amount of laser light used thus a reduction in the rate of
>> photo-toxicity.
>>
>> I hope this has helped, all the best
>>
>> steve
>>
>>
>> Steve Bagley
>> Head of Imaging
>> Imaging Facility
>> Cancer Research UK
>> Paterson Institute for Cancer Research
>> University of Manchester
>> Wilmslow Road
>> Manchester
>> M20 9BX
>> UK
>> www.paterson.man.ac.uk
>>
>>
>>
>>
>>
>>
>> ----- Original Message -----
>> From: "zhan cheng" <[hidden email]>
>> To: <[hidden email]>
>> Sent: Monday, February 22, 2010 6:50 PM
>> Subject: spinning disk confocal Vs deltavision?
>>
>>
>> Hello everyone,
>>  Our facility is planning to buy a live cell imaging setup, spinning
>> disk
>> confocal and deltavision seem both good. We are concering about  
>> imaging
>> quality, imaging speed, phototoxin, and photo bleaching for long term
>> imaging.
>> Could you share your experience? some time lapse our imaging maybe  
>> last
>> over
>> 12 hours, so  sometimes the data is huge. I know the  deltavision
>> system's
>> imaging quality is good after deconvolution, but I worry about the 3D
>> deconvolution's speed, especially for huge data.
>>   Thanks.
>>   zhan cheng
>> --------------------------------------------------------
>> This email is confidential and intended solely for the use of the  
>> person(s)
> ('the intended recipient') to whom it was addressed. Any views or  
> opinions
> presented are solely those of the author and do not necessarily  
> represent
> those of the Paterson Institute for Cancer Research or the  
> University of
> Manchester. It may contain information that is privileged &  
> confidential
> within the meaning of applicable law. Accordingly any dissemination,
> distribution, copying, or other use of this message, or any of its  
> contents,
> by any person other than the intended recipient may constitute a  
> breach of
> civil or criminal law and is strictly prohibited. If you are NOT the
> intended recipient please contact the sender and dispose of this e-
> mail as
> soon as possible.
>>

[James Pawley]

Hi all,

Thanks George. Real numbers are always good.

At 5k rpm, and 12 patterns/revolution, this seems
to add up to about 1ms/pattern. (and 10k rpm ->
0.5 ms/pattern).

Now at this point we have to think a bit about
what we mean by a pattern. I mean an array of
lenses or apertures so arranged on a spinning
disk that those just entering the imaged area as
the exposure starts will move from (say) the left
edge to the right edge and during this process
all parts of the image plane will be equally
illuminated (and recorded from).

In this case, a few of the lines of illumination
actually will go all the way from left to right,
while the majority will start from the location
where the spots were as the exposure started and
continue to the right edge. The "missing parts"
of these partial lines will be "filled in" by
illumination passing through other
lenses/apertures that are part of the incoming
pattern (the pattern just about to enter the
imaged area when the exposure began.).

If everything goes well, the exposure will stop
just as these "second pattern" spots reach the
points illuminated by the original pattern at the
start of the exposure.

If the timing is off, parts of these lines may be
either double-exposed (exposure too long) or not
exposed at all (exposure too short). In the
former case, the overlapped areas of each spot in
the array will be 2x brighter than other areas.
In the latter case, there will  be dark patches.

If the geometry of the incoming pattern does not
exactly match that of the outgoing one in the
relative size, position and distance of each spot
from the rotational axis, then again, some areas
of the field may be either double exposed or not
exposed at all.

Assuming that the patterns are indeed perfectly
aligned, avoiding line segments that are either
double-exposed or dark requires exposure timing
that is accurate to, say, about 1/500th of the
exposure time or 1-2µs. As this may be difficult
to arrange, it is more common to average the
exposures of, say, two or three patterns as under
these conditions an error results not in a
reduction or increase of 100% but one of only 50
or 33%.

Well, you can get the idea. You need more than
high rotational speed and a fast camera to get
good fast imaging. You also need a very
symmetrical, repeated pattern of "identical"
lenses or apertures and a reliable method of
synchronizing the exposure time to the period
required for a complete pattern to sweep across
the imaged area.

If you want to check this, make an image of a
specimen that is uniformly fluorescent such as a
so-called "fluorescent sea" (dye dissolved in
liquid) or a piece of fluorescent plastic.

You may be surprised what it looks like when the exposures get short.

Best..

Jim Pawley

              **********************************************
Prof. James B. Pawley,                          Ph.  608-263-3147
Room 223, Zoology Research
Building,                          FAX
608-265-5315
1117 Johnson Ave., Madison, WI, 53706  
[hidden email]
3D Microscopy of Living Cells Course, June 12-24, 2010, UBC, Vancouver Canada
Info: http://www.3dcourse.ubc.ca/             Applications due by March 15, 2010
               "If it ain't diffraction, it must be statistics." Anon.

>Actually the CSU-22 spins at 5000 rpms and csux
>up to 10,000 rpm or 2000 scans / sec (12 scans
>per rotation).
>1800 rpm (360 scans / sec) is the default speed
>that can be adjusted from software to the
>highest speed.
>
>George Peeters
>Solamere Technology Group
>
>Sent from my iPhone
>
>On Mar 1, 2010, at 12:25 PM, Andrea Latini
><[hidden email]> wrote:
>
>>Dear Dr. Zhan Cheng and dear List,
>>
>>I think you can solve all problems by using a faster spinning disk confocal
>>system, i.e. the CARV-II.
>>this is a 7500 rpm spinning disk unit and I've been able to test it either
>>with a low resolution-high sensitivity Photometrics EVOLVE 512x512 and a
>>less sensitive- higher resolution CoolSnap EZ.
>>Results:
>>with the Evolve EMCCD I've been down to 5ms exposure time without any loss
>>due to slow spinning disk speed, i.e. the CSU-22 system (this spins at less
>>than 1800rpm).
>>
>>with the CoolSnap camera system I've been able to collect very good images
>>(i.e. S/N > 5), even at 19ms exposure time.
>>
>>Of course, due to the CARV-II very high rotation speed, the only limitation
>>is from the CCD system speed.
>>
>>I'm ready to provide you with all pieces of information you may need.
>>
>>Regards.
>>
>>Andrea Latini, PhD
>>Crisel Instruments Srl, ITALY
>>[hidden email]
>>
>>On Thu, 25 Feb 2010 16:19:21 -0000, Steve
>>Bagley <[hidden email]> wrote:
>>
>>>
>>>Hi
>>>
>>>We have both a deltavision (seven years with several upgrades) and a
>>>roper csu22 spinning disk system (one year). Both have the same camera
>>>(EMCCD cascade), filters (ET-Chroma) and objective lenses
>>>
>>>In our hands we find that
>>>
>>>i) when live cell imaging fibre like structures (cytoskeletal elements)
>>>the deltavision system seems to present the better results, when imaging
>>>spots (e.g. spindle pole bodies) the spinning disk system seems more
>>>suited
>>>
>>>ii) the limiting factor on the spinning disk is exposure time. When
>>>going below 50msec the image quality drops due to the rotational speed
>>>of the disk. For very fast imaging the deltavision presents more
>>>favourable capture times. I believ that with the x1 you can go a little
>>>faster in capture speed without deterimentally effecting image quality
>>>but the scanner rotational speed is around the same.
>>>
>>>iii) overall we have seen slightly less photo-toxcity on our spinning
>>>disk system. This maybe due to the model of Olympus microscope that our
>>>systems are on (DV olympus IX71 SD olympus IX81).
>>>
>>>iv) on both systems we have full environmental chambers, bioptechs
>>>chambers, objective heaters and the cellasic microfluidic system. Both
>>>systems are well suited to enviromental control.
>>>
>>>v) I guess the draw back is the range of wavelengths available to the
>>>spinning disk via the lasers. We have the full 'sedat filter' range
>>>406/491/555/643 whereas the DV is not limited by wavelength selection.
>>>
>>>vi) with both systems the data is deconvolved afterwards, the SD to
>>>further vastly improve the axial resolution. All data goes through the
>>>same 3D imaging and analysis software. For deconvolution we have scaled
>>>up with multiprocessors and linux with Huygens, both data streams (DV
>>>and SD) can be processed this way with the same software.
>>>
>>>vii) we have been considering the Princeton ProEM 1024B to increase
>>>sensitivity and photoefficiency, however the minimum usable capture rate
>>>of the spinning disk 50msec still could not be exceeded without
>>>seriously effecting image quality. The advantage would be a reduction in
>>>the amount of laser light used thus a reduction in the rate of
>>>photo-toxicity.
>>>
>>>I hope this has helped, all the best
>>>
>>>steve
>>>
>>>
>>>Steve Bagley
>>>Head of Imaging
>>>Imaging Facility
>>>Cancer Research UK
>>>Paterson Institute for Cancer Research
>>>University of Manchester
>>>Wilmslow Road
>>>Manchester
>>>M20 9BX
>>>UK
>>>www.paterson.man.ac.uk
>>>
>>>
>>>
>>>
>>>
>>>
>>>----- Original Message -----
>>>From: "zhan cheng" <[hidden email]>
>>>To: <[hidden email]>
>>>Sent: Monday, February 22, 2010 6:50 PM
>>>Subject: spinning disk confocal Vs deltavision?
>>>
>>>
>>>Hello everyone,
>>>  Our facility is planning to buy a live cell imaging setup, spinning
>>>disk
>>>confocal and deltavision seem both good. We are concering about imaging
>>>quality, imaging speed, phototoxin, and photo bleaching for long term
>>>imaging.
>>>Could you share your experience? some time lapse our imaging maybe last
>>>over
>>>12 hours, so  sometimes the data is huge. I know the  deltavision
>>>system's
>>>imaging quality is good after deconvolution, but I worry about the 3D
>>>deconvolution's speed, especially for huge data.
>>>   Thanks.
>>>   zhan cheng
>>>--------------------------------------------------------
>>>This email is confidential and intended solely for the use of the person(s)
>>('the intended recipient') to whom it was addressed. Any views or opinions
>>presented are solely those of the author and do not necessarily represent
>>those of the Paterson Institute for Cancer Research or the University of
>>Manchester. It may contain information that is privileged & confidential
>>within the meaning of applicable law. Accordingly any dissemination,
>>distribution, copying, or other use of this message, or any of its contents,
>>by any person other than the intended recipient may constitute a breach of
>>civil or criminal law and is strictly prohibited. If you are NOT the
>>intended recipient please contact the sender and dispose of this e-mail as
>>soon as possible.

--
               **********************************************
Prof. James B. Pawley,                          Ph.  608-263-3147
Room 223, Zoology Research
Building,                          FAX
608-265-5315
1117 Johnson Ave., Madison, WI, 53706  
[hidden email]
3D Microscopy of Living Cells Course, June 12-24, 2010, UBC, Vancouver Canada
Info: http://www.3dcourse.ubc.ca/             Applications due by March 15, 2010
               "If it ain't diffraction, it must be statistics." Anon.

[Andrea Latini-2]

Jim
The situation is not exactly like that, the 12 pattern are overlapping on
the full length of the disk. So to make a single frame you still need a
complete revolution, which means that at 5000 rpm frame rate is limited to
83 FpS, at 7000 is 116 FpS.
To get a sync signal for the data acquisition could not be a problem,
however the 12 overlapping spiral are there to provide enough integration
and overlapping to make negligible non uniformities and time mismatching or
jitter, this means that you should not see artifacts even if you are not in
perfect sync with the spinning disk.
vincenzo

Vincenzo Ricco

technical director

Crisel Instruments srl

via Mattia Battistini 177

00167 Roma

ITALY

phone [phone]

fax: [phone]

[hidden email]

www.crisel-instruments.it

-----Messaggio originale-----
Da: Confocal Microscopy List [mailto:[hidden email]] Per
conto di James Pawley
Inviato: mercoledì 3 marzo 2010 23.54
A: [hidden email]
Oggetto: Re: spinning disk confocal Vs deltavision?

Hi all,

Thanks George. Real numbers are always good.

At 5k rpm, and 12 patterns/revolution, this seems
to add up to about 1ms/pattern. (and 10k rpm ->
0.5 ms/pattern).

Now at this point we have to think a bit about
what we mean by a pattern. I mean an array of
lenses or apertures so arranged on a spinning
disk that those just entering the imaged area as
the exposure starts will move from (say) the left
edge to the right edge and during this process
all parts of the image plane will be equally
illuminated (and recorded from).

In this case, a few of the lines of illumination
actually will go all the way from left to right,
while the majority will start from the location
where the spots were as the exposure started and
continue to the right edge. The "missing parts"
of these partial lines will be "filled in" by
illumination passing through other
lenses/apertures that are part of the incoming
pattern (the pattern just about to enter the
imaged area when the exposure began.).

If everything goes well, the exposure will stop
just as these "second pattern" spots reach the
points illuminated by the original pattern at the
start of the exposure.

If the timing is off, parts of these lines may be
either double-exposed (exposure too long) or not
exposed at all (exposure too short). In the
former case, the overlapped areas of each spot in
the array will be 2x brighter than other areas.
In the latter case, there will  be dark patches.

If the geometry of the incoming pattern does not
exactly match that of the outgoing one in the
relative size, position and distance of each spot
from the rotational axis, then again, some areas
of the field may be either double exposed or not
exposed at all.

Assuming that the patterns are indeed perfectly
aligned, avoiding line segments that are either
double-exposed or dark requires exposure timing
that is accurate to, say, about 1/500th of the
exposure time or 1-2µs. As this may be difficult
to arrange, it is more common to average the
exposures of, say, two or three patterns as under
these conditions an error results not in a
reduction or increase of 100% but one of only 50
or 33%.

Well, you can get the idea. You need more than
high rotational speed and a fast camera to get
good fast imaging. You also need a very
symmetrical, repeated pattern of "identical"
lenses or apertures and a reliable method of
synchronizing the exposure time to the period
required for a complete pattern to sweep across
the imaged area.

If you want to check this, make an image of a
specimen that is uniformly fluorescent such as a
so-called "fluorescent sea" (dye dissolved in
liquid) or a piece of fluorescent plastic.

You may be surprised what it looks like when the exposures get short.

Best..

Jim Pawley

              **********************************************
Prof. James B. Pawley,                          Ph.
608-263-3147
Room 223, Zoology Research
Building,                          FAX
608-265-5315
1117 Johnson Ave., Madison, WI, 53706  
[hidden email]
3D Microscopy of Living Cells Course, June 12-24, 2010, UBC, Vancouver
Canada
Info: http://www.3dcourse.ubc.ca/             Applications due by March 15,
2010
               "If it ain't diffraction, it must be statistics." Anon.

>Actually the CSU-22 spins at 5000 rpms and csux
>up to 10,000 rpm or 2000 scans / sec (12 scans
>per rotation).
>1800 rpm (360 scans / sec) is the default speed
>that can be adjusted from software to the
>highest speed.
>
>George Peeters
>Solamere Technology Group
>
>Sent from my iPhone
>
>On Mar 1, 2010, at 12:25 PM, Andrea Latini
><[hidden email]> wrote:
>
>>Dear Dr. Zhan Cheng and dear List,
>>
>>I think you can solve all problems by using a faster spinning disk

confocal
>>system, i.e. the CARV-II.
>>this is a 7500 rpm spinning disk unit and I've been able to test it either
>>with a low resolution-high sensitivity Photometrics EVOLVE 512x512 and a
>>less sensitive- higher resolution CoolSnap EZ.
>>Results:
>>with the Evolve EMCCD I've been down to 5ms exposure time without any loss
>>due to slow spinning disk speed, i.e. the CSU-22 system (this spins at
less
>>than 1800rpm).
>>
>>with the CoolSnap camera system I've been able to collect very good images
>>(i.e. S/N > 5), even at 19ms exposure time.
>>
>>Of course, due to the CARV-II very high rotation speed, the only
limitation

>>is from the CCD system speed.
>>
>>I'm ready to provide you with all pieces of information you may need.
>>
>>Regards.
>>
>>Andrea Latini, PhD
>>Crisel Instruments Srl, ITALY
>>[hidden email]
>>
>>On Thu, 25 Feb 2010 16:19:21 -0000, Steve
>>Bagley <[hidden email]> wrote:
>>
>>>
>>>Hi
>>>
>>>We have both a deltavision (seven years with several upgrades) and a
>>>roper csu22 spinning disk system (one year). Both have the same camera
>>>(EMCCD cascade), filters (ET-Chroma) and objective lenses
>>>
>>>In our hands we find that
>>>
>>>i) when live cell imaging fibre like structures (cytoskeletal elements)
>>>the deltavision system seems to present the better results, when imaging
>>>spots (e.g. spindle pole bodies) the spinning disk system seems more
>>>suited
>>>
>>>ii) the limiting factor on the spinning disk is exposure time. When
>>>going below 50msec the image quality drops due to the rotational speed
>>>of the disk. For very fast imaging the deltavision presents more
>>>favourable capture times. I believ that with the x1 you can go a little
>>>faster in capture speed without deterimentally effecting image quality
>>>but the scanner rotational speed is around the same.
>>>
>>>iii) overall we have seen slightly less photo-toxcity on our spinning
>>>disk system. This maybe due to the model of Olympus microscope that our
>>>systems are on (DV olympus IX71 SD olympus IX81).
>>>
>>>iv) on both systems we have full environmental chambers, bioptechs
>>>chambers, objective heaters and the cellasic microfluidic system. Both
>>>systems are well suited to enviromental control.
>>>
>>>v) I guess the draw back is the range of wavelengths available to the
>>>spinning disk via the lasers. We have the full 'sedat filter' range
>>>406/491/555/643 whereas the DV is not limited by wavelength selection.
>>>
>>>vi) with both systems the data is deconvolved afterwards, the SD to
>>>further vastly improve the axial resolution. All data goes through the
>>>same 3D imaging and analysis software. For deconvolution we have scaled
>>>up with multiprocessors and linux with Huygens, both data streams (DV
>>>and SD) can be processed this way with the same software.
>>>
>>>vii) we have been considering the Princeton ProEM 1024B to increase
>>>sensitivity and photoefficiency, however the minimum usable capture rate
>>>of the spinning disk 50msec still could not be exceeded without
>>>seriously effecting image quality. The advantage would be a reduction in
>>>the amount of laser light used thus a reduction in the rate of
>>>photo-toxicity.
>>>
>>>I hope this has helped, all the best
>>>
>>>steve
>>>
>>>
>>>Steve Bagley
>>>Head of Imaging
>>>Imaging Facility
>>>Cancer Research UK
>>>Paterson Institute for Cancer Research
>>>University of Manchester
>>>Wilmslow Road
>>>Manchester
>>>M20 9BX
>>>UK
>>>www.paterson.man.ac.uk
>>>
>>>
>>>
>>>
>>>
>>>
>>>----- Original Message -----
>>>From: "zhan cheng" <[hidden email]>
>>>To: <[hidden email]>
>>>Sent: Monday, February 22, 2010 6:50 PM
>>>Subject: spinning disk confocal Vs deltavision?
>>>
>>>
>>>Hello everyone,
>>>  Our facility is planning to buy a live cell imaging setup, spinning
>>>disk
>>>confocal and deltavision seem both good. We are concering about imaging
>>>quality, imaging speed, phototoxin, and photo bleaching for long term
>>>imaging.
>>>Could you share your experience? some time lapse our imaging maybe last
>>>over
>>>12 hours, so  sometimes the data is huge. I know the  deltavision
>>>system's
>>>imaging quality is good after deconvolution, but I worry about the 3D
>>>deconvolution's speed, especially for huge data.
>>>   Thanks.
>>>   zhan cheng
>>>--------------------------------------------------------
>>>This email is confidential and intended solely for the use of the

person(s)
>>('the intended recipient') to whom it was addressed. Any views or opinions
>>presented are solely those of the author and do not necessarily represent
>>those of the Paterson Institute for Cancer Research or the University of
>>Manchester. It may contain information that is privileged & confidential
>>within the meaning of applicable law. Accordingly any dissemination,
>>distribution, copying, or other use of this message, or any of its
contents,
>>by any person other than the intended recipient may constitute a breach of
>>civil or criminal law and is strictly prohibited. If you are NOT the
>>intended recipient please contact the sender and dispose of this e-mail as
>>soon as possible.


--
               **********************************************
Prof. James B. Pawley,                          Ph.
608-263-3147
Room 223, Zoology Research
Building,                          FAX
608-265-5315
1117 Johnson Ave., Madison, WI, 53706  
[hidden email]
3D Microscopy of Living Cells Course, June 12-24, 2010, UBC, Vancouver
Canada
Info: http://www.3dcourse.ubc.ca/             Applications due by March 15,
2010
               "If it ain't diffraction, it must be statistics." Anon.

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[Axel Kurt Preuss]

My experience is that a CHANGE in camera speed creates short lived
disturbances ( in non or not well  synched) systems , not so less the
absolute speed per se.
This should be easy to test by checking whether only speed increase or
both increase and decrease manifest themselves as disturbances
Cheers
Ax


Thnx
Regards

Axel

IMCB-Central Imaging
[phone]

On Mar 4, 2010, at 5:30 PM, "Ricco" <[hidden email]> wrote:

> Jim
> The situation is not exactly like that, the 12 pattern are
> overlapping on
> the full length of the disk. So to make a single frame you still
> need a
> complete revolution, which means that at 5000 rpm frame rate is
> limited to
> 83 FpS, at 7000 is 116 FpS.
> To get a sync signal for the data acquisition could not be a problem,
> however the 12 overlapping spiral are there to provide enough
> integration
> and overlapping to make negligible non uniformities and time
> mismatching or
> jitter, this means that you should not see artifacts even if you are
> not in
> perfect sync with the spinning disk.
> vincenzo
>
> Vincenzo Ricco
>
> technical director
>
> Crisel Instruments srl
>
> via Mattia Battistini 177
>
> 00167 Roma
>
> ITALY
>
> phone [phone]
>
> fax: [phone]
>
> [hidden email]
>
> www.crisel-instruments.it
>
> -----Messaggio originale-----
> Da: Confocal Microscopy List
> [mailto:[hidden email]] Per
> conto di James Pawley
> Inviato: mercoledì 3 marzo 2010 23.54
> A: [hidden email]
> Oggetto: Re: spinning disk confocal Vs deltavision?
>
> Hi all,
>
> Thanks George. Real numbers are always good.
>
> At 5k rpm, and 12 patterns/revolution, this seems
> to add up to about 1ms/pattern. (and 10k rpm ->
> 0.5 ms/pattern).
>
> Now at this point we have to think a bit about
> what we mean by a pattern. I mean an array of
> lenses or apertures so arranged on a spinning
> disk that those just entering the imaged area as
> the exposure starts will move from (say) the left
> edge to the right edge and during this process
> all parts of the image plane will be equally
> illuminated (and recorded from).
>
> In this case, a few of the lines of illumination
> actually will go all the way from left to right,
> while the majority will start from the location
> where the spots were as the exposure started and
> continue to the right edge. The "missing parts"
> of these partial lines will be "filled in" by
> illumination passing through other
> lenses/apertures that are part of the incoming
> pattern (the pattern just about to enter the
> imaged area when the exposure began.).
>
> If everything goes well, the exposure will stop
> just as these "second pattern" spots reach the
> points illuminated by the original pattern at the
> start of the exposure.
>
> If the timing is off, parts of these lines may be
> either double-exposed (exposure too long) or not
> exposed at all (exposure too short). In the
> former case, the overlapped areas of each spot in
> the array will be 2x brighter than other areas.
> In the latter case, there will  be dark patches.
>
> If the geometry of the incoming pattern does not
> exactly match that of the outgoing one in the
> relative size, position and distance of each spot
> from the rotational axis, then again, some areas
> of the field may be either double exposed or not
> exposed at all.
>
> Assuming that the patterns are indeed perfectly
> aligned, avoiding line segments that are either
> double-exposed or dark requires exposure timing
> that is accurate to, say, about 1/500th of the
> exposure time or 1-2µs. As this may be difficult
> to arrange, it is more common to average the
> exposures of, say, two or three patterns as under
> these conditions an error results not in a
> reduction or increase of 100% but one of only 50
> or 33%.
>
> Well, you can get the idea. You need more than
> high rotational speed and a fast camera to get
> good fast imaging. You also need a very
> symmetrical, repeated pattern of "identical"
> lenses or apertures and a reliable method of
> synchronizing the exposure time to the period
> required for a complete pattern to sweep across
> the imaged area.
>
> If you want to check this, make an image of a
> specimen that is uniformly fluorescent such as a
> so-called "fluorescent sea" (dye dissolved in
> liquid) or a piece of fluorescent plastic.
>
> You may be surprised what it looks like when the exposures get short.
>
> Best..
>
> Jim Pawley
>
>              **********************************************
> Prof. James B. Pawley,                                   Ph.
> 608-263-3147
> Room 223, Zoology Research
> Building,                              FAX
> 608-265-5315
> 1117 Johnson Ave., Madison, WI, 53706
> [hidden email]
> 3D Microscopy of Living Cells Course, June 12-24, 2010, UBC, Vancouver
> Canada
> Info: http://www.3dcourse.ubc.ca/         Applications due by March
> 15,
> 2010
>           "If it ain't diffraction, it must be statistics." Anon.
>
>> Actually the CSU-22 spins at 5000 rpms and csux
>> up to 10,000 rpm or 2000 scans / sec (12 scans
>> per rotation).
>> 1800 rpm (360 scans / sec) is the default speed
>> that can be adjusted from software to the
>> highest speed.
>>
>> George Peeters
>> Solamere Technology Group
>>
>> Sent from my iPhone
>>
>> On Mar 1, 2010, at 12:25 PM, Andrea Latini
>> <[hidden email]> wrote:
>>
>>> Dear Dr. Zhan Cheng and dear List,
>>>
>>> I think you can solve all problems by using a faster spinning disk
> confocal
>>> system, i.e. the CARV-II.
>>> this is a 7500 rpm spinning disk unit and I've been able to test
>>> it either
>>> with a low resolution-high sensitivity Photometrics EVOLVE 512x512
>>> and a
>>> less sensitive- higher resolution CoolSnap EZ.
>>> Results:
>>> with the Evolve EMCCD I've been down to 5ms exposure time without
>>> any loss
>>> due to slow spinning disk speed, i.e. the CSU-22 system (this
>>> spins at
> less
>>> than 1800rpm).
>>>
>>> with the CoolSnap camera system I've been able to collect very
>>> good images
>>> (i.e. S/N > 5), even at 19ms exposure time.
>>>
>>> Of course, due to the CARV-II very high rotation speed, the only
> limitation
>>> is from the CCD system speed.
>>>
>>> I'm ready to provide you with all pieces of information you may
>>> need.
>>>
>>> Regards.
>>>
>>> Andrea Latini, PhD
>>> Crisel Instruments Srl, ITALY
>>> [hidden email]
>>>
>>> On Thu, 25 Feb 2010 16:19:21 -0000, Steve
>>> Bagley <[hidden email]> wrote:
>>>
>>>>
>>>> Hi
>>>>
>>>> We have both a deltavision (seven years with several upgrades)
>>>> and a
>>>> roper csu22 spinning disk system (one year). Both have the same
>>>> camera
>>>> (EMCCD cascade), filters (ET-Chroma) and objective lenses
>>>>
>>>> In our hands we find that
>>>>
>>>> i) when live cell imaging fibre like structures (cytoskeletal
>>>> elements)
>>>> the deltavision system seems to present the better results, when
>>>> imaging
>>>> spots (e.g. spindle pole bodies) the spinning disk system seems
>>>> more
>>>> suited
>>>>
>>>> ii) the limiting factor on the spinning disk is exposure time. When
>>>> going below 50msec the image quality drops due to the rotational
>>>> speed
>>>> of the disk. For very fast imaging the deltavision presents more
>>>> favourable capture times. I believ that with the x1 you can go a
>>>> little
>>>> faster in capture speed without deterimentally effecting image
>>>> quality
>>>> but the scanner rotational speed is around the same.
>>>>
>>>> iii) overall we have seen slightly less photo-toxcity on our
>>>> spinning
>>>> disk system. This maybe due to the model of Olympus microscope
>>>> that our
>>>> systems are on (DV olympus IX71 SD olympus IX81).
>>>>
>>>> iv) on both systems we have full environmental chambers, bioptechs
>>>> chambers, objective heaters and the cellasic microfluidic system.
>>>> Both
>>>> systems are well suited to enviromental control.
>>>>
>>>> v) I guess the draw back is the range of wavelengths available to
>>>> the
>>>> spinning disk via the lasers. We have the full 'sedat filter' range
>>>> 406/491/555/643 whereas the DV is not limited by wavelength
>>>> selection.
>>>>
>>>> vi) with both systems the data is deconvolved afterwards, the SD to
>>>> further vastly improve the axial resolution. All data goes
>>>> through the
>>>> same 3D imaging and analysis software. For deconvolution we have
>>>> scaled
>>>> up with multiprocessors and linux with Huygens, both data streams
>>>> (DV
>>>> and SD) can be processed this way with the same software.
>>>>
>>>> vii) we have been considering the Princeton ProEM 1024B to increase
>>>> sensitivity and photoefficiency, however the minimum usable
>>>> capture rate
>>>> of the spinning disk 50msec still could not be exceeded without
>>>> seriously effecting image quality. The advantage would be a
>>>> reduction in
>>>> the amount of laser light used thus a reduction in the rate of
>>>> photo-toxicity.
>>>>
>>>> I hope this has helped, all the best
>>>>
>>>> steve
>>>>
>>>>
>>>> Steve Bagley
>>>> Head of Imaging
>>>> Imaging Facility
>>>> Cancer Research UK
>>>> Paterson Institute for Cancer Research
>>>> University of Manchester
>>>> Wilmslow Road
>>>> Manchester
>>>> M20 9BX
>>>> UK
>>>> www.paterson.man.ac.uk
>>>>
>>>>
>>>>
>>>>
>>>>
>>>>
>>>> ----- Original Message -----
>>>> From: "zhan cheng" <[hidden email]>
>>>> To: <[hidden email]>
>>>> Sent: Monday, February 22, 2010 6:50 PM
>>>> Subject: spinning disk confocal Vs deltavision?
>>>>
>>>>
>>>> Hello everyone,
>>>> Our facility is planning to buy a live cell imaging setup, spinning
>>>> disk
>>>> confocal and deltavision seem both good. We are concering about
>>>> imaging
>>>> quality, imaging speed, phototoxin, and photo bleaching for long
>>>> term
>>>> imaging.
>>>> Could you share your experience? some time lapse our imaging
>>>> maybe last
>>>> over
>>>> 12 hours, so  sometimes the data is huge. I know the  deltavision
>>>> system's
>>>> imaging quality is good after deconvolution, but I worry about
>>>> the 3D
>>>> deconvolution's speed, especially for huge data.
>>>>  Thanks.
>>>>  zhan cheng
>>>> --------------------------------------------------------
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