High speed spinning disc confocal with EMCCD camera

[sivaram mylavarapu]

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Dear colleagues,

We are planning on acquiring a high-speed, high-sensitivity confocal
fluorescence imaging station for meeting two simultaneous needs: 1) high
speed for live cell imaging and 2) very high sensitivity to be able to
capture very low light images. We have been seriously considering a
spinning disc confocal with an EMCCD camera attachment to meet both these
needs. We would of course include halogen illumination for routine
widefield fluorescence imaging.

I wanted to request your opinions on the following:

1) Which is the best microscope in the market for these needs? I have used
a Zeiss spinning disc in the past, and it appears Leica has an instrument
too - both use Yokogawa spinning discs. Nikon sells their system with an
Andor spinning disc.

2) Which is the best EMCCD camera for such a system?

3) Can I combine the above features to be supplied with a dual inverted and
upright microscope so that both high resolution live cell (inverted) and C
elegans/ Zebrafish/ Drosophila embryo live imaging (upright) can be
performed as per need?

I would appreciate your valuable inputs!

Warm regards,

Sivaram.
--
Sivaram Mylavarapu.
[hidden email]

[John Oreopoulos]

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Dear Sivaram,

In regards to point 3) that you raised, for such a setup, I think you'll need two spinning disk scan heads... Although with a fancy set of switchable image relays, you might be able to share one scan head for the upright and inverted sections of the microscope. Never seen anything like that attempted though.

John Oreopoulos


On 2014-12-16, at 8:17 PM, sivaram mylavarapu wrote:

> *****
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> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear colleagues,
>
> We are planning on acquiring a high-speed, high-sensitivity confocal
> fluorescence imaging station for meeting two simultaneous needs: 1) high
> speed for live cell imaging and 2) very high sensitivity to be able to
> capture very low light images. We have been seriously considering a
> spinning disc confocal with an EMCCD camera attachment to meet both these
> needs. We would of course include halogen illumination for routine
> widefield fluorescence imaging.
>
> I wanted to request your opinions on the following:
>
> 1) Which is the best microscope in the market for these needs? I have used
> a Zeiss spinning disc in the past, and it appears Leica has an instrument
> too - both use Yokogawa spinning discs. Nikon sells their system with an
> Andor spinning disc.
>
> 2) Which is the best EMCCD camera for such a system?
>
> 3) Can I combine the above features to be supplied with a dual inverted and
> upright microscope so that both high resolution live cell (inverted) and C
> elegans/ Zebrafish/ Drosophila embryo live imaging (upright) can be
> performed as per need?
>
> I would appreciate your valuable inputs!
>
> Warm regards,
>
> Sivaram.
> --
> Sivaram Mylavarapu.
> [hidden email]

[Rosemary.White]

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Hi Sivaram,

You might also consider purchasing two separate systems, a spinning disc
system for your sensitive imaging, as described, and a build-it-yourself
SPIM system for the live embryos. A PhD student here is planning to
assemble of these "suitcase SPIMs" as described here:
http://openspim.org/Welcome_to_the_OpenSPIM_Wiki. You have to buy (mostly)
or make the components, but from the prices listed, it seems quite cheap
and there are detailed instructions. If you your someone in your lab is
keen, it'd be a fun project.

just a thought...
cheers,
Rosemary

Dr Rosemary White
CSIRO Black Mountain
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
E [hidden email]


On 17/12/14 12:17 PM, "sivaram mylavarapu" <[hidden email]> wrote:

>*****
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>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>Post images on http://www.imgur.com and include the link in your posting.
>*****
>
>Dear colleagues,
>
>We are planning on acquiring a high-speed, high-sensitivity confocal
>fluorescence imaging station for meeting two simultaneous needs: 1) high
>speed for live cell imaging and 2) very high sensitivity to be able to
>capture very low light images. We have been seriously considering a
>spinning disc confocal with an EMCCD camera attachment to meet both these
>needs. We would of course include halogen illumination for routine
>widefield fluorescence imaging.
>
>I wanted to request your opinions on the following:
>
>1) Which is the best microscope in the market for these needs? I have used
>a Zeiss spinning disc in the past, and it appears Leica has an instrument
>too - both use Yokogawa spinning discs. Nikon sells their system with an
>Andor spinning disc.
>
>2) Which is the best EMCCD camera for such a system?
>
>3) Can I combine the above features to be supplied with a dual inverted
>and
>upright microscope so that both high resolution live cell (inverted) and C
>elegans/ Zebrafish/ Drosophila embryo live imaging (upright) can be
>performed as per need?
>
>I would appreciate your valuable inputs!
>
>Warm regards,
>
>Sivaram.
>--
>Sivaram Mylavarapu.
>[hidden email]

[George McNamara]

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Hi Sivaram,

Consider sCMOS instead of EMCCD. PCO may have a small hardware edge over
Andor and Hamamatsu when using the 4.2 sensor. My Hamamatsu FLASH4
system (MetaMorph 7.8.x) has flaky crashing issues - I will not be
surprised if this is due to the D-CAM driver being poorly written (and
suspect PCO is far better).

Consider the spidery platform on the right side of the instrument
pictured near the bottom of
http://www.biovis.com/diskovery.htm

Also consider the less expensive (and does not require lasers) Crisel
X-Light (3rd generation what started out as Atto CARV) and whatever LED
fluorescence illumination the vendor recommends ... 2 X-Lights + 2 LED
fl illuminators will probably cost less than a Yokogawa+laser stack.
http://www.biovis.com/x-light.htm

//

"We would of course include halogen illumination for routine widefield
fluorescence imaging."

Really?

//

spinning disks are so 1996 (maybe 1997 - first gen Yokogawa CSU-10 in
Shinya Inoue's lab, MBL, see for a 12512322 later publication). At least
Olympus FV-OSR and Zeiss AiryScan are pushing point scanning confocals
(and then there is STED - too bad Leica has crappy print ads for it that
leave the side by side images looking pretty much identical).

For speed and resolution (better than any confocal, or FV-OSR or
AiryScan), email or talk with Hari about iSIM:

*Instant* super-resolution imaging in live cells and embryos via analog
image processing. <http://www.ncbi.nlm.nih.gov/pubmed/24097271>

*York* AG, Chandris P, Nogare DD, Head J, Wawrzusin P, Fischer RS,
Chitnis A, *Shroff*H.

Nat Methods. 2013; 10: 1122-6. doi: 10.1038/nmeth.2687. Epub 2013 Oct 6.

PMID:
    24097271



//

Speaking of lasers - only two weeks left before
http://www.wickedlasers.com/ stops selling >5 mW lasers in the USA ...
reminder: do not look at laser with remaining eye.
They apparently will still sell Torch lamp to burn things
http://www.wickedlasers.com/torch

Enjoy,

George

On 12/16/2014 7:17 PM, sivaram mylavarapu wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear colleagues,
>
> We are planning on acquiring a high-speed, high-sensitivity confocal
> fluorescence imaging station for meeting two simultaneous needs: 1) high
> speed for live cell imaging and 2) very high sensitivity to be able to
> capture very low light images. We have been seriously considering a
> spinning disc confocal with an EMCCD camera attachment to meet both these
> needs. We would of course include halogen illumination for routine
> widefield fluorescence imaging.
>
> I wanted to request your opinions on the following:
>
> 1) Which is the best microscope in the market for these needs? I have used
> a Zeiss spinning disc in the past, and it appears Leica has an instrument
> too - both use Yokogawa spinning discs. Nikon sells their system with an
> Andor spinning disc.
>
> 2) Which is the best EMCCD camera for such a system?
>
> 3) Can I combine the above features to be supplied with a dual inverted and
> upright microscope so that both high resolution live cell (inverted) and C
> elegans/ Zebrafish/ Drosophila embryo live imaging (upright) can be
> performed as per need?
>
> I would appreciate your valuable inputs!
>
> Warm regards,
>
> Sivaram.
>    

--



George McNamara, Ph.D.
Single Cells Analyst
L.J.N. Cooper Lab
University of Texas M.D. Anderson Cancer Center
Houston, TX 77054
Tattletales http://works.bepress.com/gmcnamara/42

[Andreas Bruckbauer]

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 If bleaching and photo damage is an issue (as usually with live cell imaging), check out this high tech solution by the Betzig group:

http://www.sciencemag.org/content/346/6208/1257998.full
it should be commercially available from 3i
https://www.intelligent-imaging.com/systems.php#lattice

 
best wishes

Andreas


 

-----Original Message-----
From: sivaram mylavarapu <[hidden email]>
To: CONFOCALMICROSCOPY <[hidden email]>
Sent: Wed, 17 Dec 2014 1:32
Subject: High speed spinning disc confocal with EMCCD camera


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*****

Dear colleagues,

We are planning on acquiring a high-speed, high-sensitivity confocal
fluorescence imaging station for meeting two simultaneous needs: 1) high
speed for live cell imaging and 2) very high sensitivity to be able to
capture very low light images. We have been seriously considering a
spinning disc confocal with an EMCCD camera attachment to meet both these
needs. We would of course include halogen illumination for routine
widefield fluorescence imaging.

I wanted to request your opinions on the following:

1) Which is the best microscope in the market for these needs? I have used
a Zeiss spinning disc in the past, and it appears Leica has an instrument
too - both use Yokogawa spinning discs. Nikon sells their system with an
Andor spinning disc.

2) Which is the best EMCCD camera for such a system?

3) Can I combine the above features to be supplied with a dual inverted and
upright microscope so that both high resolution live cell (inverted) and C
elegans/ Zebrafish/ Drosophila embryo live imaging (upright) can be
performed as per need?

I would appreciate your valuable inputs!

Warm regards,

Sivaram.
--
Sivaram Mylavarapu.
[hidden email]

 

[Philippe clemenceau]

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COMMERCIAL POST :

For the community using spinning disk confocal,  I wanted to communicate that there is now an even faster option ( up to several KHz frame rate)  for truly high speed spinning disk confocal , using a high speed , cooled intensified CMOS camera.

We just launched the HICAM FLUO from Lambert Instruments which outputs 1 Mpixels at 1 KHz with cooled Gen 3 intensifier.

Please check http://www.lambertinstruments.com/hicam-fluo <http://www.lambertinstruments.com/hicam-fluo>

This would be interesting for researchers who need to observe very fast phenomenon (ms scale) or perform extremely fast 3D scanning .

Kind regards,

Philippe Clémenceau,
Co-Founder, MS in Optical Science
Axiom Optics
 
 
1 Broadway, 14th floor
Cambridge, MA 02142, USA
 
Phone:  [phone]
Mobile: [phone]

Check out our product line at www.axiomoptics.com <http://www.axiomoptics.com/>   :

-Scientific Imaging (CCD, EMCCD, SCMOS, SWIR, CMOS, Intensified )
-Laser Beam profilers (UV , NIR,  IR)
-Advanced Light Microscopy  ( Fluorescence Lifetime, Phase Imaging  & Adaptive Optics)

> On Dec 17, 2014, at 4:24 AM, Andreas Bruckbauer <[hidden email]> wrote:
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
>
> If bleaching and photo damage is an issue (as usually with live cell imaging), check out this high tech solution by the Betzig group:
>
> http://www.sciencemag.org/content/346/6208/1257998.full
> it should be commercially available from 3i
> https://www.intelligent-imaging.com/systems.php#lattice
>
>
> best wishes
>
> Andreas
>
>
>
>
> -----Original Message-----
> From: sivaram mylavarapu <[hidden email]>
> To: CONFOCALMICROSCOPY <[hidden email]>
> Sent: Wed, 17 Dec 2014 1:32
> Subject: High speed spinning disc confocal with EMCCD camera
>
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear colleagues,
>
> We are planning on acquiring a high-speed, high-sensitivity confocal
> fluorescence imaging station for meeting two simultaneous needs: 1) high
> speed for live cell imaging and 2) very high sensitivity to be able to
> capture very low light images. We have been seriously considering a
> spinning disc confocal with an EMCCD camera attachment to meet both these
> needs. We would of course include halogen illumination for routine
> widefield fluorescence imaging.
>
> I wanted to request your opinions on the following:
>
> 1) Which is the best microscope in the market for these needs? I have used
> a Zeiss spinning disc in the past, and it appears Leica has an instrument
> too - both use Yokogawa spinning discs. Nikon sells their system with an
> Andor spinning disc.
>
> 2) Which is the best EMCCD camera for such a system?
>
> 3) Can I combine the above features to be supplied with a dual inverted and
> upright microscope so that both high resolution live cell (inverted) and C
> elegans/ Zebrafish/ Drosophila embryo live imaging (upright) can be
> performed as per need?
>
> I would appreciate your valuable inputs!
>
> Warm regards,
>
> Sivaram.
> --
> Sivaram Mylavarapu.
> [hidden email]
>
>
>
>

[Andrea Latini]

*****
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Post images on http://www.imgur.com and include the link in your posting.
*****

COMMERCIAL RESPONSE:

Dear George, thank you for bringing to Sivaram attention the option to use
our X-Light Confocal System. this is actually the only system capable of
20KRPM spinning disk rotation speed and proprietary spiral pattern to avoid
the needs to detector/disk sync down to 1ms exposure time to avoid spiral
striping.

As you can easily understand any detector synchronization with disk position
will force you to slow down your maximum frame rate according to the sync
overlap between detector exposure+readout time and disk rotation.

for any further information please get in touch, directly.

best.

Andrea Latini
CrestOptics srl
phone: [phone]
[hidden email]
www.crestopt.com

[David Burgess]

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*****

we had access for two months last summer to the Brucker Opterra.  It is
exceedingly fast, we saw exceedingly low toxicity relative to our spinning
disc for live cell work on three different species of embryos, and very
controllable.  Just viewed their totally upgraded software package at the
ASCB meeting making the system easy to use and ideal for cell biology and
neuroscience. The price point is really competitive and better than most
commercial systems.

David Burgess
National Research Mentoring Network, Lead PI
Boston College
Professor
Biology Department
528 Higgins Hall
140 Commonwealth Ave
Chestnut Hill, MA 02467

617-552-1606


On Wed, Dec 17, 2014 at 8:44 AM, Andrea Latini <[hidden email]> wrote:

>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> COMMERCIAL RESPONSE:
>
> Dear George, thank you for bringing to Sivaram attention the option to use
> our X-Light Confocal System. this is actually the only system capable of
> 20KRPM spinning disk rotation speed and proprietary spiral pattern to avoid
> the needs to detector/disk sync down to 1ms exposure time to avoid spiral
> striping.
>
> As you can easily understand any detector synchronization with disk
> position
> will force you to slow down your maximum frame rate according to the sync
> overlap between detector exposure+readout time and disk rotation.
>
> for any further information please get in touch, directly.
>
> best.
>
> Andrea Latini
> CrestOptics srl
> phone: [phone]
> [hidden email]
> www.crestopt.com
>

[Watkins, Simon C]

Reflecting on Davids comment.  We use the Optera extensively... we have 6 of them.. However  all  are run under NIS elements not the Bruker/Prairie software.  We also have 2 Yokagawa scan heads which we rarely use because of the vast difference in performance and flexibility.  The Optera scan head has many advantages over the spinning disk approach.  Firstly the core technology is an array of pinholes or slits which means you can change between multiple different pinholes and slits (7 in total) to adapt confocality to objective NA and specimen brightness (in other words you can go fast imaging dim specimens if you are willing to make compromises in confocality).   Secondly it is not just a disk spinning in a box.   When it is appropriately integrated with piezo Z, TTL control and a decent laser launch it becomes an incredibly powerful imaging platform.  As I said I cannot comment on the current version of the Bruker/PrairieView software as we are firmly embedded in the Nikon platform which runs it beautifully and flexibly, but as a high speed confocal I do not think there is any other real competition.

Simon C. Watkins Ph.D, FRC Path
Distinguished Professor and Vice Chair Cell Biology
Professor Immunology
Director Center for Biologic Imaging
BSTS 225
University of Pittsburgh
3500 Terrace St
Pittsburgh PA 15261
412-352-2277
www.cbi.pitt.edu

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of David Burgess
Sent: Wednesday, December 17, 2014 10:17 AM
To: [hidden email]
Subject: Re: High speed spinning disc confocal with EMCCD camera

*****
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we had access for two months last summer to the Brucker Opterra.  It is exceedingly fast, we saw exceedingly low toxicity relative to our spinning disc for live cell work on three different species of embryos, and very controllable.  Just viewed their totally upgraded software package at the ASCB meeting making the system easy to use and ideal for cell biology and neuroscience. The price point is really competitive and better than most commercial systems.

David Burgess
National Research Mentoring Network, Lead PI Boston College Professor Biology Department
528 Higgins Hall
140 Commonwealth Ave
Chestnut Hill, MA 02467

617-552-1606


On Wed, Dec 17, 2014 at 8:44 AM, Andrea Latini <[hidden email]> wrote:

>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> COMMERCIAL RESPONSE:
>
> Dear George, thank you for bringing to Sivaram attention the option to
> use our X-Light Confocal System. this is actually the only system
> capable of 20KRPM spinning disk rotation speed and proprietary spiral
> pattern to avoid the needs to detector/disk sync down to 1ms exposure
> time to avoid spiral striping.
>
> As you can easily understand any detector synchronization with disk
> position will force you to slow down your maximum frame rate according
> to the sync overlap between detector exposure+readout time and disk
> rotation.
>
> for any further information please get in touch, directly.
>
> best.
>
> Andrea Latini
> CrestOptics srl
> phone: [phone]
> [hidden email]
> www.crestopt.com
>

[samjlord]

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On Wed, 17 Dec 2014 06:47:02 +0530, sivaram mylavarapu
<[hidden email]> wrote:
>1) Which is the best microscope in the market for these needs? I have used
>a Zeiss spinning disc in the past, and it appears Leica has an instrument
>too - both use Yokogawa spinning discs. Nikon sells their system with an
>Andor spinning disc.

I've used both the Yokagawa CSU-X and the Andor/Spectral Diskovery system.
Both on a Nikon Ti-E base. They are comparable in their throughput. The Diskovery
is much more flexible, and we have it with two camera ports with an EMCCD and a
CMOS. Also, we can perform confocal and TIRF onto the same cameras without
moving them. The downside is more complexity, larger footprint, and high cost
(although the price is approximately the same as if you were going to buy Nikon
TIRF, Yokogawa disk, and lamp/LED epi).

I would highly recommend either of these systems, especially if your lab has the
expertise to set up TTL triggering. Neither is exactly "turn-key," but you get
flexibility.

In order to perform the different imaging modes that you mentioned, you'll either
have to get the Diskovery (which can move the disk out of the way) or simply
move the camera from the spinning disk port to another port on the microscope.
For the latter option, you'll need a separate excitation source, like a lamp or LED
epi illuminator (I love the Lumencor Spectra-X, but there are many good options).
If you need laser illumination, such as for TIRF, you'll need a laser combiner that
has a second fiber output.

>2) Which is the best EMCCD camera for such a system?

I've used Andor iXons (both the 512 and the 1024 pixel versions) as well as CMOS
(Hamamatsu Flash4 and Andor Zyla 4.2).

I would recommend a 512 EMCCD for spinning disk and single-molecule imaging.
The flexibility and sensitivity of an EMCCD just isn't yet matched with CMOS. And
the new EMCCDs run at very fast frame rates. That said, CMOS is cheaper, higher
resolution, as fast fast fast. I love our CMOS cameras for epi, TIRF, and z-stacks of
bright samples. For single-molecule, EMCCD is a hair better. (Some of that has to
do with the larger pixels on the EMCCD. So if your CMOS supports binning, that's
an option. Also, if you don't mag the detection as much, you can get that dim
signal into fewer pixels and boost your signal to noise, but then you lose the
benefit of the high-res camera.)

Most of the companies really use the same sensor, so it matters more that you get
the best price, that you get good support, and that the camera works well with
your software. For these reasons, I typically go with Andor, but Photometrics,
Hamamatsu, etc. are all excellent cameras. (I find that Andor support on Micro-
Manager is superior.)

>3) Can I combine the above features to be supplied with a dual inverted and
>upright microscope so that both high resolution live cell (inverted) and C
>elegans/ Zebrafish/ Drosophila embryo live imaging (upright) can be
>performed as per need?

I'm not 100% sure what you intend to do there. Do you want diSPIM? Nikon and
other companies are starting to integrate that:
http://www.nikoninstruments.com/About-Nikon/News-Room/US-News/Nikon-
Instruments-Inc.-Unveils-New-Ti-diSPIM-System-at-Cell-Biology-2014

You should be able to add that on top of a spinning disk scope.

I'd be happy to discuss more. Just send me a personal message.

-Sam

[Yoann Gosselin]

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COMMERCIAL POST:

Dear Sivaram,

Imaging with EMCCD cameras offers the best sensitivity in low-light imaging. When background signals contaminating the image are not the limiting factor, the noise generated by the camera starts having a significant impact on the image quality.

The three main specifications when comparing EMCCDs for spinning disk are speed, dark current (or thermal noise) and clock-induced charges (CIC, also called spurious background). All high grade EMCCD camera manufacturers use the same e2v sensors, so have roughly the same quantum efficiency (that can will vary slightly depending on the coating). The main differences between cameras are with the quality of the vacuum, the thermoelectric cooling (TEC) and the electronics. The vacuum and TEC will influence the dark current. Only cameras with high quality vacuum and TEC will reach a stabilized temperature of -85 +/- 0.01°C with air cooling at 20MHz. Dark current roughly doubles with every 7°C. The EM gain is extremely sensitive to temperature variations, so a stabilized cooling is key for quality images. Better electronics will minimize CIC, which is the dominant source of noise in low-light imaging with EMCCDs.

You can find more details on EMCCDs in low light imaging here:

http://labrigger.com/blog/2013/01/16/emccd-specs/
http://www.photonics.com/Article.aspx?AID=56742

As for the comparison with sCMOS, you will find that EMCCD cameras, with their larger pixels, higher quantum efficiency and negligible readout noise, are more sensitive than sCMOS in low-light imaging. The main advantage of using sCMOS cameras is the pixel readout speed. Most EMCCD cameras now read at approx. 20MHz pixel readout rate, which gives ~60 fps with a 512 x 512 camera. A sCMOS camera can reach 100 fps with a 2048 x 2048 sensor. However, the faster you image with a camera, the lower the exposure time, which means that you need more sensitivity to get quality images. That is why a lot of sCMOS users do not use the max. speed of their sCMOS camera while using binning, which decrease the spatial resolution of your sensor.

Feel free to contact me with any question you might have on the matter. I would be happy to discuss this in more details.

Regards,

Yoann Gosselin, B.Eng., M.A.Sc.
Ingénieur en applications
Applications Engineer
Nüvü Caméras Inc.
+1.514.733.8666, ext. 1019
nuvucameras.com

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*********************************

[Zdenek Svindrych-2]

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Hi Sivaram,
I have two additional short comments:

1) Andor uses Yokogawa spinning discs as well.

2) You may also consider structured illumination-based optical sectioning
microscopes. It's nothing to do with superresolution but the sectioning can
in fact be better than with a confocal (http://www.opticsinfobase.org/oe/
abstract.cfm?uri=oe-20-22-24585). Old Zeiss ApoTome worked that way. Now
there are better units out there, such as Zeiss VivaTome or Andor Revolution
DSD, that don't do any complicated image processing and try to capture all
the fluorescence coming from your sample (to be fair, there is some
subtraction of fluorescence images, which worsens the SNR somewhat).

I have tried the Andor unit (no commercial interest), they sell it as a
'budget confocal', but I was quite impressed with the results... Also it's
pretty compact and uses arc lamp (or LED) illumination.

Best, zdenek






---------- Původní zpráva ----------
Od: sivaram mylavarapu <[hidden email]>
Komu: [hidden email]
Datum: 16. 12. 2014 20:33:21
Předmět: High speed spinning disc confocal with EMCCD camera

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Dear colleagues,

We are planning on acquiring a high-speed, high-sensitivity confocal
fluorescence imaging station for meeting two simultaneous needs: 1) high
speed for live cell imaging and 2) very high sensitivity to be able to
capture very low light images. We have been seriously considering a
spinning disc confocal with an EMCCD camera attachment to meet both these
needs. We would of course include halogen illumination for routine
widefield fluorescence imaging.

I wanted to request your opinions on the following:

1) Which is the best microscope in the market for these needs? I have used
a Zeiss spinning disc in the past, and it appears Leica has an instrument
too - both use Yokogawa spinning discs. Nikon sells their system with an
Andor spinning disc.

2) Which is the best EMCCD camera for such a system?

3) Can I combine the above features to be supplied with a dual inverted and
upright microscope so that both high resolution live cell (inverted) and C
elegans/ Zebrafish/ Drosophila embryo live imaging (upright) can be
performed as per need?

I would appreciate your valuable inputs!

Warm regards,

Sivaram.
--
Sivaram Mylavarapu.
[hidden email]"

[lechristophe]

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Hi,

I think that now that they aquired Spectral Applied Research, Andor sells
the Diskovery system (different from the Revolution), which is not Yokogawa
based?

Christophe

Le 19 déc. 2014 03:48, "Zdenek Svindrych" <[hidden email]> a écrit :

▶ Afficher le texte des messages précédents

[Kurt Thorn]

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On 12/19/2014 1:29 AM, Christophe Leterrier wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi,
>
> I think that now that they aquired Spectral Applied Research, Andor sells
> the Diskovery system (different from the Revolution), which is not Yokogawa
> based?

My understanding is that Andor sells both Yokogawa systems and the
Diskovery system. In fact, I believe there will be a Borealis
modification to the CSU-W1 available soon from Andor / Spectral Applied
Research.

Kurt

>
> Christophe
>
> Le 19 déc. 2014 03:48, "Zdenek Svindrych" <[hidden email]> a écrit :
>
> ▶ Afficher le texte des messages précédents
>
>


--
Kurt Thorn
Associate Professor
Director, Nikon Imaging Center
http://thornlab.ucsf.edu/
http://nic.ucsf.edu/blog/

[John Oreopoulos]

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*** Commercial response ***

Kurt, Christophe, you are both correct. Just some additional clarification and information:

Spectral Applied Research was acquired by Andor Technologies in October of 2013. The Diskovery Platform is a new spinning disk confocal system designed and manufactured in Canada and was officially "re-launched" by Andor at this year's annual Society for Neuroscience Meeting. This instrument has a number of distinguishing features - the application of the Borealis illumination scheme, removable and exchangeable confocal pinhole disk cartridges with different pinhole sizes and spacings (to change the degree of confocality and pinhole cross-talk), sCMOS camera compatible field of view, and an upgradable TIRF module that integrates into the same optical pathway as the confocal imaging mode thus allowing one to use the same camera for confocal and TIRF imaging (with automatic registration of the images). These and other additional features are described in the product brochure on the Andor website.


John Oreopoulos
Staff Scientist
Spectral Applied Research Inc.
A Division of Andor Technology
Richmond Hill, Ontario
Canada
www.spectral.ca


On 2014-12-19, at 12:27 PM, Kurt Thorn wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> On 12/19/2014 1:29 AM, Christophe Leterrier wrote:
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> Post images on http://www.imgur.com and include the link in your posting.
>> *****
>>
>> Hi,
>>
>> I think that now that they aquired Spectral Applied Research, Andor sells
>> the Diskovery system (different from the Revolution), which is not Yokogawa
>> based?
> My understanding is that Andor sells both Yokogawa systems and the Diskovery system. In fact, I believe there will be a Borealis modification to the CSU-W1 available soon from Andor / Spectral Applied Research.
>
> Kurt
>
>>
>> Christophe
>>
>> Le 19 déc. 2014 03:48, "Zdenek Svindrych" <[hidden email]> a écrit :
>>
>> ▶ Afficher le texte des messages précédents
>>
>>
>
>
> --
> Kurt Thorn
> Associate Professor
> Director, Nikon Imaging Center
> http://thornlab.ucsf.edu/
> http://nic.ucsf.edu/blog/

[Marco De Spirito]

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Hi Sivaram,

we have bough the Crisel  X-Light one years ago.  Image quality is
comparable, if not better, than those of standard laser confocal systems,
and the price is really competitive. Furthermore the software (MetaMorph)
does not undergo to any crash (at least up to now!!).

Now I know they have introduced a new version with added fast structured
illumination for up to 80-100 namometers resolution and I’m really curious
to see how it works.

Best,

Marco

2014-12-21 4:56 GMT+01:00 John Oreopoulos <[hidden email]>:

>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> *** Commercial response ***
>
> Kurt, Christophe, you are both correct. Just some additional clarification
> and information:
>
> Spectral Applied Research was acquired by Andor Technologies in October of
> 2013. The Diskovery Platform is a new spinning disk confocal system
> designed and manufactured in Canada and was officially "re-launched" by
> Andor at this year's annual Society for Neuroscience Meeting. This
> instrument has a number of distinguishing features - the application of the
> Borealis illumination scheme, removable and exchangeable confocal pinhole
> disk cartridges with different pinhole sizes and spacings (to change the
> degree of confocality and pinhole cross-talk), sCMOS camera compatible
> field of view, and an upgradable TIRF module that integrates into the same
> optical pathway as the confocal imaging mode thus allowing one to use the
> same camera for confocal and TIRF imaging (with automatic registration of
> the images). These and other additional features are described in the
> product brochure on the Andor website.
>
>
> John Oreopoulos
> Staff Scientist
> Spectral Applied Research Inc.
> A Division of Andor Technology
> Richmond Hill, Ontario
> Canada
> www.spectral.ca
>
>
> On 2014-12-19, at 12:27 PM, Kurt Thorn wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > On 12/19/2014 1:29 AM, Christophe Leterrier wrote:
> >> *****
> >> To join, leave or search the confocal microscopy listserv, go to:
> >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> >> Post images on http://www.imgur.com and include the link in your
> posting.
> >> *****
> >>
> >> Hi,
> >>
> >> I think that now that they aquired Spectral Applied Research, Andor
> sells
> >> the Diskovery system (different from the Revolution), which is not
> Yokogawa
> >> based?
> > My understanding is that Andor sells both Yokogawa systems and the
> Diskovery system. In fact, I believe there will be a Borealis modification
> to the CSU-W1 available soon from Andor / Spectral Applied Research.
> >
> > Kurt
> >
> >>
> >> Christophe
> >>
> >> Le 19 déc. 2014 03:48, "Zdenek Svindrych" <[hidden email]> a écrit :
> >>
> >> ▶ Afficher le texte des messages précédents
> >>
> >>
> >
> >
> > --
> > Kurt Thorn
> > Associate Professor
> > Director, Nikon Imaging Center
> > http://thornlab.ucsf.edu/
> > http://nic.ucsf.edu/blog/
>

--
Marco De Spirito
Università Cattolica S. Cuore
Istituto di Fisica
L.go F. Vito 1
00168 - Roma - ITALY

ph.: + 39 06 30154265
fax: + 39 06 3013858
e-mail: [hidden email]
website: http://docenti.unicatt.it/ita/marco_de_spirito/
social 1: https://www.researchgate.net/profile/Marco_De_Spirito/
social 2: http://scholar.google.it/citations?user=EiJux1sAAAAJ
skype: mictomchia

[Andrea Latini]

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- 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]

[Andrew York]

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 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://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com 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]
>

[Andrea Latini]

*****
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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://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>Post images on http://www.imgur.com 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://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> Post images on http://www.imgur.com 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]
>>

[Feinstein, Timothy]

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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: [phone] | 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]
>>>