Spinning disk comparison - CSU-X1 vs CrestOptics X-Light V3

[Giang, William]

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Dear list,

Has anyone compared the emitted light gathering capabilities of a Yokogawa CSU-X1 vs a CrestOptics X-Light V3?
Neither have a microlens array on the emission path, and both units would have 50um diameter pinholes with 250um spacing.

Techs from a large microscopy company said the CSU-X1 is superior for fast live-cell imaging in cell culture, but it's not clear to me why. Is Yokogawa able to get a higher density of pinholes on their disk? Am I misunderstanding how pinhole spacing works?

x-posted to Microforum at https://forum.microlist.org/t/yokogawa-csu-x1-vs-crest-x-light-v3-light-output/1240

Thanks in advance,
Will


William Giang  |  Imaging Specialist

Whitehead Building, Room 465
615 Michael Street, Atlanta, GA 30322
[hidden email]<mailto:[hidden email]>
ici.emory.edu<http://www.cores.emory.edu/ici/>
(304) 853-DATA


Remember to acknowledge ICI in your publications.<https://www.cores.emory.edu/ici/Acknowledgements/index.html>



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[Dr. K N Ganesh]

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Hi
1. CSU X1 being dual disc spinning system has micro lenses on excitation side  that enhances the excitation efficiency
2.CSUX1 is laser based has an ability to synchronised disc rotation with respect  exposure time of camera  . (1800 rpm  or 5000 rpm )
Whereas crest offers simple single Nipkow Disc without micro lenses and more importantly it uses bright LED light if I am not wrong.
Naturally CSUX1 should perform better.  
Regards
Ganesh

Sent from my iPhone

> On 19-Dec-2020, at 05:34, Giang, William <[hidden email]> wrote:
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
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> *****
>
> Dear list,
>
> Has anyone compared the emitted light gathering capabilities of a Yokogawa CSU-X1 vs a CrestOptics X-Light V3?
> Neither have a microlens array on the emission path, and both units would have 50um diameter pinholes with 250um spacing.
>
> Techs from a large microscopy company said the CSU-X1 is superior for fast live-cell imaging in cell culture, but it's not clear to me why. Is Yokogawa able to get a higher density of pinholes on their disk? Am I misunderstanding how pinhole spacing works?
>
> x-posted to Microforum at https://forum.microlist.org/t/yokogawa-csu-x1-vs-crest-x-light-v3-light-output/1240
>
> Thanks in advance,
> Will
>
>
> William Giang  |  Imaging Specialist
>
> Whitehead Building, Room 465
> 615 Michael Street, Atlanta, GA 30322
> [hidden email]<mailto:[hidden email]>
> ici.emory.edu<http://www.cores.emory.edu/ici/>
> (304) 853-DATA
>
>
> Remember to acknowledge ICI in your publications.<https://www.cores.emory.edu/ici/Acknowledgements/index.html>
>
>
>
> ________________________________
>
> This e-mail message (including any attachments) is for the sole use of
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[Alessandra Scarpellini]

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**COMMERCIAL REPLY**

Dear Confocal List,

I'd like to point out a few key points on the CrestOptics solutions, especially the X-light V3:
- Yes, there are no micro-lenses to focus excitation light, however this is compensated by the use of more powerful laser sources (cost-effective multi-mode sources) to achieve comparable excitation power on the sample
- On The X-light V3 micro-lenses are employed to achieve homogeneous illumination, so that quantitative imaging can be done on the full field of view (FOV). This FOV measures 25mm, meaning a much faster data collection (more cells in a single FOV)
- 2 cameras with FOV 25mm can be used simultaneously for even faster data collection
- CrestOptics disk spins at 15K RPM, meaning acquisition speed >1kHz without artefacts
- Disk pattern can be customized for the best ratio of confocality and throughput depending on the sample

These and many more info can be found on our website: www.crestoptics.com

Best regards,

Alessandra



Alessandra Scarpellini, PhD | Head of Sales and Marketing
Via di Torre Rossa, 66 00165 ROME - Italy
M: [phone] P: [phone]
 
   
 
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-----Messaggio originale-----
Da: Confocal Microscopy List <[hidden email]> Per conto di Ganesh Kadasoor
Inviato: sabato 19 dicembre 2020 11:29
A: [hidden email]
Oggetto: Re: Spinning disk comparison - CSU-X1 vs CrestOptics X-Light V3

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Hi
1. CSU X1 being dual disc spinning system has micro lenses on excitation side  that enhances the excitation efficiency
2.CSUX1 is laser based has an ability to synchronised disc rotation with respect  exposure time of camera  . (1800 rpm  or 5000 rpm ) Whereas crest offers simple single Nipkow Disc without micro lenses and more importantly it uses bright LED light if I am not wrong.
Naturally CSUX1 should perform better.  
Regards
Ganesh

Sent from my iPhone

> On 19-Dec-2020, at 05:34, Giang, William <[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.
> *****
>
> Dear list,
>
> Has anyone compared the emitted light gathering capabilities of a Yokogawa CSU-X1 vs a CrestOptics X-Light V3?
> Neither have a microlens array on the emission path, and both units would have 50um diameter pinholes with 250um spacing.
>
> Techs from a large microscopy company said the CSU-X1 is superior for fast live-cell imaging in cell culture, but it's not clear to me why. Is Yokogawa able to get a higher density of pinholes on their disk? Am I misunderstanding how pinhole spacing works?
>
> x-posted to Microforum at
> https://forum.microlist.org/t/yokogawa-csu-x1-vs-crest-x-light-v3-ligh
> t-output/1240
>
> Thanks in advance,
> Will
>
>
> William Giang  |  Imaging Specialist
>
> Whitehead Building, Room 465
> 615 Michael Street, Atlanta, GA 30322
> [hidden email]<mailto:[hidden email]>
> ici.emory.edu<http://www.cores.emory.edu/ici/>
> (304) 853-DATA
>
>
> Remember to acknowledge ICI in your
> publications.<https://www.cores.emory.edu/ici/Acknowledgements/index.h
> tml>
>
>
>
> ________________________________
>
> This e-mail message (including any attachments) is for the sole use of
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> recipient, you are hereby notified that any dissemination,
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> strictly prohibited.
>
> If you have received this message in error, please contact the sender
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[Knecht, David]

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I have never understood the rationale for micro-lenses in a spinning disk system.  These systems are aimed at live cell imaging.  I have never used anywhere near full laser power on either our Yokogawa system or our Crest system.  Cells do not tolerate high excitation light in general and so we always work with the minimum light level  possible and that is never near the limit of laser power.  So I don’t see the rationale for the added cost of a micro-lens system, unless you have a really low power laser source.  I would be curioius to know if anyone has a different opinion.  As to the question that was asked, I have not directly compared the two, but not sure how I would since they are on different microscopes, objectives, cameras, laser launches, etc.  I can say that both work to give nice images of live cells.  I like the ability of the Crest to function in confocal or non-confocal mode which our Yokogawa system cannot do.  Dave

On Dec 19, 2020, at 8:16 AM, Alessandra Scarpellini <[hidden email]<mailto:[hidden email]>> wrote:

*Message sent from a system outside of UConn.*


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**COMMERCIAL REPLY**

Dear Confocal List,

I'd like to point out a few key points on the CrestOptics solutions, especially the X-light V3:
- Yes, there are no micro-lenses to focus excitation light, however this is compensated by the use of more powerful laser sources (cost-effective multi-mode sources) to achieve comparable excitation power on the sample
- On The X-light V3 micro-lenses are employed to achieve homogeneous illumination, so that quantitative imaging can be done on the full field of view (FOV). This FOV measures 25mm, meaning a much faster data collection (more cells in a single FOV)
- 2 cameras with FOV 25mm can be used simultaneously for even faster data collection
- CrestOptics disk spins at 15K RPM, meaning acquisition speed >1kHz without artefacts
- Disk pattern can be customized for the best ratio of confocality and throughput depending on the sample

These and many more info can be found on our website: https://nam10.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.crestoptics.com%2F&amp;data=04%7C01%7Cdavid.knecht%40UCONN.EDU%7Cf930fd5e77584f2e2ce408d8a421ca48%7C17f1a87e2a254eaab9df9d439034b080%7C0%7C0%7C637439812353517251%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000&amp;sdata=0D05rdwBnXFYFyNOFfiJEvODpD4QF1vJksr%2B%2FDPGuhU%3D&amp;reserved=0

Best regards,

Alessandra



Alessandra Scarpellini, PhD | Head of Sales and Marketing
Via di Torre Rossa, 66 00165 ROME - Italy
M: [phone] P: [phone]



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-----Messaggio originale-----
Da: Confocal Microscopy List <[hidden email]> Per conto di Ganesh Kadasoor
Inviato: sabato 19 dicembre 2020 11:29
A: [hidden email]
Oggetto: Re: Spinning disk comparison - CSU-X1 vs CrestOptics X-Light V3

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Hi
1. CSU X1 being dual disc spinning system has micro lenses on excitation side  that enhances the excitation efficiency
2.CSUX1 is laser based has an ability to synchronised disc rotation with respect  exposure time of camera  . (1800 rpm  or 5000 rpm ) Whereas crest offers simple single Nipkow Disc without micro lenses and more importantly it uses bright LED light if I am not wrong.
Naturally CSUX1 should perform better.
Regards
Ganesh

Sent from my iPhone

On 19-Dec-2020, at 05:34, Giang, William <[hidden email]> wrote:

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Dear list,

Has anyone compared the emitted light gathering capabilities of a Yokogawa CSU-X1 vs a CrestOptics X-Light V3?
Neither have a microlens array on the emission path, and both units would have 50um diameter pinholes with 250um spacing.

Techs from a large microscopy company said the CSU-X1 is superior for fast live-cell imaging in cell culture, but it's not clear to me why. Is Yokogawa able to get a higher density of pinholes on their disk? Am I misunderstanding how pinhole spacing works?

x-posted to Microforum at
https://nam10.safelinks.protection.outlook.com/?url=https%3A%2F%2Fforum.microlist.org%2Ft%2Fyokogawa-csu-x1-vs-crest-x-light-v3-ligh&amp;data=04%7C01%7Cdavid.knecht%40UCONN.EDU%7Cf930fd5e77584f2e2ce408d8a421ca48%7C17f1a87e2a254eaab9df9d439034b080%7C0%7C0%7C637439812353517251%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000&amp;sdata=aeFi3tg5EB3Je7jnM7rA1BRZGE3ytQyQPqQFwJ9WnIs%3D&amp;reserved=0
t-output/1240

Thanks in advance,
Will


William Giang  |  Imaging Specialist

Whitehead Building, Room 465
615 Michael Street, Atlanta, GA 30322
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[Zdenek Svindrych-2]

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Hi David,

the rationale is simple. The purpose of the spinning disk is to attenuate
out-of-focus light. If the CSU-X1 attenuates this unwanted emission by a
factor of 30 (estimate, I didn't do the math), it will also attenuate the
laser excitation by the same factor. With a strong laser that shouldn't be
a problem, right?
Well, 1:30 pinhole crosstalk may not be enough. If you bleach a (small,
let's say 1 um by 1 um) cone of light in a thick fluorescent layer you
won't be able to see it with the X1, but you can see it clearly with a
point scanner. That's why CSU-W1 or Dragonfly have sparser pinholes and
higher attenuation. Without the microlenses you would end up with very
little excitation even with powerful lasers.
And as a matter of fact, we use quite strong excitation (30% with a 150 mW
laser) to capture a z-stack quickly and do longer pauses between z-stacks -
this helps with motion artifacts and allows for deconvolution...

Back to the original question - with identical pinhole pattern and
overall optical configuration (minus the excitation pinholes), the
detection efficiency should be the same. The ultimate limiting factor will
be how much light can the disk handle, how you deal with the reflected
laser light from the disk (essentially 100%), and the autofluorescence of
any element that is common to the strong excitation (before the disk) and
emission paths...

Best, zdenek

[Craig Brideau]

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As Zdenek says, you can need quite substantial laser powers under some
scenarios without the microlenses concentrating the energy. The cost for
high-power versions of certain lasers can also be quite high, if the
required energy density is available at all, so on occasion a weaker laser
is your only option. A lower power laser also requires less heat sinking
and electrical current, which simplifies the overall design and in some
cases can lead to longer laser life if thermal design is still taken
seriously by the manufacturer.

Craig

On Sun, Dec 20, 2020 at 11:28 AM Zdenek Svindrych <[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.
> *****
>
> Hi David,
>
> the rationale is simple. The purpose of the spinning disk is to attenuate
> out-of-focus light. If the CSU-X1 attenuates this unwanted emission by a
> factor of 30 (estimate, I didn't do the math), it will also attenuate the
> laser excitation by the same factor. With a strong laser that shouldn't be
> a problem, right?
> Well, 1:30 pinhole crosstalk may not be enough. If you bleach a (small,
> let's say 1 um by 1 um) cone of light in a thick fluorescent layer you
> won't be able to see it with the X1, but you can see it clearly with a
> point scanner. That's why CSU-W1 or Dragonfly have sparser pinholes and
> higher attenuation. Without the microlenses you would end up with very
> little excitation even with powerful lasers.
> And as a matter of fact, we use quite strong excitation (30% with a 150 mW
> laser) to capture a z-stack quickly and do longer pauses between z-stacks -
> this helps with motion artifacts and allows for deconvolution...
>
> Back to the original question - with identical pinhole pattern and
> overall optical configuration (minus the excitation pinholes), the
> detection efficiency should be the same. The ultimate limiting factor will
> be how much light can the disk handle, how you deal with the reflected
> laser light from the disk (essentially 100%), and the autofluorescence of
> any element that is common to the strong excitation (before the disk) and
> emission paths...
>
> Best, zdenek
>

[Giang, William]

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Ganesh - #2 - I've encountered no issues with hardware triggering on a Nikon scope with the X-Light V2 LFOV, and the light engine was the Lumencor CELESTA with 1W laser lines. Thanks Alessandra for covering #1.

Dave - I concur. I've never wanted/needed more power in the context of live cell imaging, and I've been pleased with the V2 LFOV. If you had a power meter that could ensure the same power density at the sample plane, I'd love to see what the difference between your systems would be. But it does sound like there are too many things that'd have to be estimated to make the comparison conclusive.

Zdenek - Yes, I've heard many people suggesting at least 100mW lasers for the W1. Some have also suggested (for when you're fine with diffraction limited resolution) using the SoRa microlensed emission disk with 1x mag to collect roughly 3x the light vs the standard 50um W1 disk. But I'd need more than a couple $600 relief checks for the SoRa disk.

For the X-Light V2 LFOV, I believe the emission filters have high (6+) OD to block (reflected) laser light from making it to the detector. I never really considered if this reflected laser light would excite out-of-focus regions of the sample--and either increase background or induce unnecessary photodamage--but maybe there's a little bit of that going on? I suppose the V3's software controlled square iris also partially sidesteps the issue of cranking up the laser power.

Craig - As Alessandra said, they use multimode fiber coupled light engines (Lumencor CELESTA or 89 North LDI) which are cheaper than the Yokogawa SDs which require single mode optical fiber input. I'm still shocked that Andrew Seeber got "an x-light V3 with the LDI from 89North, two Hamamatsu Fusion cameras, all the optics I need, a VisiView license, a second hand Zeiss Observer 7 body and a table for ~$200k. (source at https://forum.microlist.org/t/crest-x-light-v3-spinning-disk-confocal-reviews/565/15). Really not sure how he swung that.

I promise this conversation was started in good faith and that I'm not a shill!

Thanks all for your responses.

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Craig Brideau
Sent: Sunday, December 20, 2020 5:49 PM
To: [hidden email]
Subject: [External] Re: Spinning disk comparison - CSU-X1 vs CrestOptics X-Light V3

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As Zdenek says, you can need quite substantial laser powers under some scenarios without the microlenses concentrating the energy. The cost for high-power versions of certain lasers can also be quite high, if the required energy density is available at all, so on occasion a weaker laser is your only option. A lower power laser also requires less heat sinking and electrical current, which simplifies the overall design and in some cases can lead to longer laser life if thermal design is still taken seriously by the manufacturer.

Craig

On Sun, Dec 20, 2020 at 11:28 AM Zdenek Svindrych <[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.
> *****
>
> Hi David,
>
> the rationale is simple. The purpose of the spinning disk is to
> attenuate out-of-focus light. If the CSU-X1 attenuates this unwanted
> emission by a factor of 30 (estimate, I didn't do the math), it will
> also attenuate the laser excitation by the same factor. With a strong
> laser that shouldn't be a problem, right?
> Well, 1:30 pinhole crosstalk may not be enough. If you bleach a
> (small, let's say 1 um by 1 um) cone of light in a thick fluorescent
> layer you won't be able to see it with the X1, but you can see it
> clearly with a point scanner. That's why CSU-W1 or Dragonfly have
> sparser pinholes and higher attenuation. Without the microlenses you
> would end up with very little excitation even with powerful lasers.
> And as a matter of fact, we use quite strong excitation (30% with a
> 150 mW
> laser) to capture a z-stack quickly and do longer pauses between
> z-stacks - this helps with motion artifacts and allows for deconvolution...
>
> Back to the original question - with identical pinhole pattern and
> overall optical configuration (minus the excitation pinholes), the
> detection efficiency should be the same. The ultimate limiting factor
> will be how much light can the disk handle, how you deal with the
> reflected laser light from the disk (essentially 100%), and the
> autofluorescence of any element that is common to the strong
> excitation (before the disk) and emission paths...
>
> Best, zdenek
>

________________________________

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[leavesley]

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Hi All,

One limitation we have run into with the CSU (we have a W1) is the
availability of suitable short-pass dichroic beamsplitters with
sufficient flatness (I guess the option to use standard long-pass
dichroics would be a benefit of the Crest system).  We are working on a
custom configuration of the CSU where we are trying to find short-pass
dichroics with edge cut-offs at 500, 550, or 600 nm.  Does anyone know
of suppliers who might be able to meet this need?  Thanks!

Best regards,

Silas


On 12/20/2020 4:49 PM, Craig Brideau 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.
> *****
>
> As Zdenek says, you can need quite substantial laser powers under some
> scenarios without the microlenses concentrating the energy. The cost for
> high-power versions of certain lasers can also be quite high, if the
> required energy density is available at all, so on occasion a weaker laser
> is your only option. A lower power laser also requires less heat sinking
> and electrical current, which simplifies the overall design and in some
> cases can lead to longer laser life if thermal design is still taken
> seriously by the manufacturer.
>
> Craig
>
> On Sun, Dec 20, 2020 at 11:28 AM Zdenek Svindrych <[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.
>> *****
>>
>> Hi David,
>>
>> the rationale is simple. The purpose of the spinning disk is to attenuate
>> out-of-focus light. If the CSU-X1 attenuates this unwanted emission by a
>> factor of 30 (estimate, I didn't do the math), it will also attenuate the
>> laser excitation by the same factor. With a strong laser that shouldn't be
>> a problem, right?
>> Well, 1:30 pinhole crosstalk may not be enough. If you bleach a (small,
>> let's say 1 um by 1 um) cone of light in a thick fluorescent layer you
>> won't be able to see it with the X1, but you can see it clearly with a
>> point scanner. That's why CSU-W1 or Dragonfly have sparser pinholes and
>> higher attenuation. Without the microlenses you would end up with very
>> little excitation even with powerful lasers.
>> And as a matter of fact, we use quite strong excitation (30% with a 150 mW
>> laser) to capture a z-stack quickly and do longer pauses between z-stacks -
>> this helps with motion artifacts and allows for deconvolution...
>>
>> Back to the original question - with identical pinhole pattern and
>> overall optical configuration (minus the excitation pinholes), the
>> detection efficiency should be the same. The ultimate limiting factor will
>> be how much light can the disk handle, how you deal with the reflected
>> laser light from the disk (essentially 100%), and the autofluorescence of
>> any element that is common to the strong excitation (before the disk) and
>> emission paths...
>>
>> Best, zdenek
>>

--


Silas J. Leavesley, Ph.D.

Professor
Department of Chemical and Biomolecular Engineering
Department of Pharmacology
Center for Lung Biology
University of South Alabama
150 Student Services Drive, SH4129
Mobile, AL 36688
ph: (251)-460-6160
fax: [phone]
web: http://www.southalabama.edu/centers/bioimaging 
<http://www.southalabama.edu/centers/bioimaging>
<http://www.usahealthsystem.com/clb>

[Mark Cannell-2]

*****
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*****

Hi Silas

Chroma can make such filter with very high performance on flat substrates but for one offs it' an expensive route... Can you use a suitable wide band pass filter such as their https://www.chroma.com/products/parts/et510-80m which is a short pass at 475-500 ? Note that the filter will tune to a shorter wavelength if used at 45 degrees and Chroma support will be able to advise the best choice from their existing filters (No commercial interest -but tell them I sent you __).

HTH Mark

Mark B. Cannell. Ph.D. FRSNZ FISHR
Department of Physiology, Pharmacology & Neuroscience
School of Medical Sciences
University Walk
Bristol BS8 1TD

 

On 7/01/21, 10:31 PM, "Confocal Microscopy List on behalf of Silas Leavesley" <[hidden email] on behalf of [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.
    *****

    Hi All,

    One limitation we have run into with the CSU (we have a W1) is the
    availability of suitable short-pass dichroic beamsplitters with
    sufficient flatness (I guess the option to use standard long-pass
    dichroics would be a benefit of the Crest system).  We are working on a
    custom configuration of the CSU where we are trying to find short-pass
    dichroics with edge cut-offs at 500, 550, or 600 nm.  Does anyone know
    of suppliers who might be able to meet this need?  Thanks!

    Best regards,

    Silas


    On 12/20/2020 4:49 PM, Craig Brideau 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.
    > *****
    >
    > As Zdenek says, you can need quite substantial laser powers under some
    > scenarios without the microlenses concentrating the energy. The cost for
    > high-power versions of certain lasers can also be quite high, if the
    > required energy density is available at all, so on occasion a weaker laser
    > is your only option. A lower power laser also requires less heat sinking
    > and electrical current, which simplifies the overall design and in some
    > cases can lead to longer laser life if thermal design is still taken
    > seriously by the manufacturer.
    >
    > Craig
    >
    > On Sun, Dec 20, 2020 at 11:28 AM Zdenek Svindrych <[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.
    >> *****
    >>
    >> Hi David,
    >>
    >> the rationale is simple. The purpose of the spinning disk is to attenuate
    >> out-of-focus light. If the CSU-X1 attenuates this unwanted emission by a
    >> factor of 30 (estimate, I didn't do the math), it will also attenuate the
    >> laser excitation by the same factor. With a strong laser that shouldn't be
    >> a problem, right?
    >> Well, 1:30 pinhole crosstalk may not be enough. If you bleach a (small,
    >> let's say 1 um by 1 um) cone of light in a thick fluorescent layer you
    >> won't be able to see it with the X1, but you can see it clearly with a
    >> point scanner. That's why CSU-W1 or Dragonfly have sparser pinholes and
    >> higher attenuation. Without the microlenses you would end up with very
    >> little excitation even with powerful lasers.
    >> And as a matter of fact, we use quite strong excitation (30% with a 150 mW
    >> laser) to capture a z-stack quickly and do longer pauses between z-stacks -
    >> this helps with motion artifacts and allows for deconvolution...
    >>
    >> Back to the original question - with identical pinhole pattern and
    >> overall optical configuration (minus the excitation pinholes), the
    >> detection efficiency should be the same. The ultimate limiting factor will
    >> be how much light can the disk handle, how you deal with the reflected
    >> laser light from the disk (essentially 100%), and the autofluorescence of
    >> any element that is common to the strong excitation (before the disk) and
    >> emission paths...
    >>
    >> Best, zdenek
    >>
    --


    Silas J. Leavesley, Ph.D.

    Professor
    Department of Chemical and Biomolecular Engineering
    Department of Pharmacology
    Center for Lung Biology
    University of South Alabama
    150 Student Services Drive, SH4129
    Mobile, AL 36688
    ph: (251)-460-6160
    fax: [phone]
    web: http://www.southalabama.edu/centers/bioimaging 
    <http://www.southalabama.edu/centers/bioimaging>
    <http://www.usahealthsystem.com/clb>