Posted by
Andrew York on
URL: http://confocal-microscopy-list.275.s1.nabble.com/High-speed-spinning-disc-confocal-with-EMCCD-camera-tp7583142p7583207.html
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I'm guessing the z-piezo is the speed limiting factor for most systems, but
let's assume everyone's using Wilson-style remote refocusing and changing
z-planes is really fast. With tons of illumination power, I'd bet the next
speed limiting factor is signal starvation due to saturation of
fluorescence excitation, so total speed is going to be proportional to the
volume of the sample that's giving useful signal. Single-point-scanning
confocal will be slowest, widefield/SPIM (with a true sheet) will be
fastest, and parallel-point-scanning/line-scanning/SPIM (with a digitally
scanned pencil to make a sheet) will be in between. SPIM gets bonus points
since it's not generating any "bad" signal to degrade the SNR of the "good"
signal, and widefield gets a penalty in most samples due to poor sectioning
degrading in-focus SNR.
Now let's suppose the sample is really densely tagged, so saturation isn't
limiting. I'd be surprised if scan speed were a major limit for anything
but non-resonant single-point confocal, so next important limiting factor
is probably data flow rate. Confocal, SPIM, and instant SIM-type methods
measure one pixel per voxel. Digital-postprocessing SIM-type methods
measure ~15 pixels per voxel, and Airyscan measures ~30. Typical computers
choke on much more than 1 GB/s, so this is probably the practical limit for
most techniques. I'm only experienced with sCMOS/EMCCD cameras, so I'm
assuming single-pixel detectors for confocal can spit out tons of data per
second; please let me know if I'm wrong about this.
On Sat, Jan 3, 2015 at 5:12 PM, Feinstein, Timothy <
[hidden email]> wrote:
> *****
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> *****
>
> Hi all,
>
> Does anyone think it would be possible to tabulate a ¹speed limit' for the
> various options discussed? I know it sounds near impossible to come up
> with a standard basis for comparison, but let¹s say something
> approximating a 512x512 acquisition either fixed or or a volume that
> includes 10 z steps (e.g., using a piezo stage when relevant). It would
> be great to have an order of magnitude idea how to compare technologies
> like a resonant scanner, Optera-type swept field scanner, spinning disc,
> VCS super-spinning disc or light sheet instrument when FPS is a major
> priority and excitation light is not limiting. Maybe we could crowdsource
> it from what users actually get in practice.
>
> All the best,
>
>
> Tim
>
> Timothy Feinstein, Ph.D. | Manager, Core for
> Confocal Microscopy and Quantitative Imaging
> 333 Bostwick Ave., N.E., Grand Rapids, Michigan 49503
> Phone: 616-234-5819 | Email:
[hidden email]
>
>
>
>
>
>
>
> On 12/30/14, 2:36 AM, "Andrea Latini" <
[hidden email]> wrote:
>
> >*****
> >To join, leave or search the confocal microscopy listserv, go to:
> >
>
http://scanmail.trustwave.com/?c=129&d=09ai1Hwf389A_JSBGnaEcBqKN1nFKNqOL1R> >ax-qpJw&u=http%3a%2f%2flists%2eumn%2eedu%2fcgi-bin%2fwa%3fA0%3dconfocalmic
> >roscopy
> >Post images on
> >
>
http://scanmail.trustwave.com/?c=129&d=09ai1Hwf389A_JSBGnaEcBqKN1nFKNqOLwF> >bleD9dw&u=http%3a%2f%2fwww%2eimgur%2ecom and include the link in your
> >posting.
> >*****
> >
> >Dear Andrew,
> >the VCS (Video Confocal Super Resolution), module is an X-Light Spinning
> >disk system
> >add-on.
> >the disk is out of the optical path when in VCS mode (i.e. 'bypass' mode).
> >basically, it's a new implementation of structured illumination
> >technology aimed to fast
> >image acquisition with no resolution limitations that are spinning disk
> >related.
> >
> >I'll be pleased to discuss more, please get in touch.
> >
> >Regards.
> >
> >Andrea
> >
[hidden email]
> >
> >
> >On Mon, 29 Dec 2014 16:58:15 -0500, Andrew York
> ><
[hidden email]> wrote:
> >
> >>*****
> >>To join, leave or search the confocal microscopy listserv, go to:
> >>
>
http://scanmail.trustwave.com/?c=129&d=09ai1Hwf389A_JSBGnaEcBqKN1nFKNqOL1> >>Rax-qpJw&u=http%3a%2f%2flists%2eumn%2eedu%2fcgi-bin%2fwa%3fA0%3dconfocalm
> >>icroscopy
> >>Post images on
> >>
>
http://scanmail.trustwave.com/?c=129&d=09ai1Hwf389A_JSBGnaEcBqKN1nFKNqOLw> >>FbleD9dw&u=http%3a%2f%2fwww%2eimgur%2ecom and include the link in your
> >>posting.
> >>*****
> >>
> >> Is there information available about this product? Is this an
> >>implementation of Enderlein's spinning disk paper? Also, 80 nm seems...
> >>optimistic? Is this with very short wavelength light, or just a slightly
> >>different definition of resolution than I'm used to?
> >>
> >>On Mon, Dec 29, 2014 at 4:10 PM, Andrea Latini <
[hidden email]>
> >>wrote:
> >>
> >>> *****
> >>> To join, leave or search the confocal microscopy listserv, go to:
> >>>
> >>>
>
http://scanmail.trustwave.com/?c=129&d=09ai1Hwf389A_JSBGnaEcBqKN1nFKNqOL> >>>1Rax-qpJw&u=http%3a%2f%2flists%2eumn%2eedu%2fcgi-bin%2fwa%3fA0%3dconfoca
> >>>lmicroscopy
> >>> Post images on
> >>>
>
http://scanmail.trustwave.com/?c=129&d=09ai1Hwf389A_JSBGnaEcBqKN1nFKNqOL> >>>wFbleD9dw&u=http%3a%2f%2fwww%2eimgur%2ecom and include the link in your
> >>>posting.
> >>> *****
> >>>
> >>> - commercial response
> >>>
> >>> thanks for reporting your experience with our Confocals Marco.
> >>>
> >>> the new Video Super Resolution module for XLight allows for 50ms
> >>>exposure
> >>> time and <1
> >>> second, 80nm spatial resolution; this is possible with large Cuda
> >>> programming we've been
> >>> developing during past months and introduced @SfN 2014 as a product.
> >>>
> >>> soon on our website and in your Lab, hopefully!
> >>>
> >>> Cheers.
> >>>
> >>> Andrea
> >>>
> >>> CrestOptics
> >>>
[hidden email]
> >>>
>