Sheppard's paper on Resolution - wall removed

[Alberto Diaspro]

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Dear
following a discussion with the MRT staff and Wiley, they informed me that the paper is now available for free for one month. Hope this can help.
Please, have a check and let me know if it works at http://onlinelibrary.wiley.com/doi/10.1002/jemt.22834/full

Best
Alby

[James Pawley]

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Hi all,

Those attending the sadly-gone UBC 3D-course may remember that our first  slogan was, "If it isn’t diffraction, it’s statistics.”

Colin’s wonderful, inclusive paper does indeed wrap up the diffraction part superbly.

But how many photons/nm*3 would you have to collect in order to obtain data that would allow you to fill in those beautifully narrow lines? Millions? (if the line width is say 1% of its “value” surely you need 10,000 detected photons to determine it  And how big must a pixel be to trace out these lovely curves without them looking blocky? Given that biologists habitually work with specimens capable of producing at most tens of detectable photons/pixel, surely it is the statistics that places the most severe spatial limits on the data we finally obtain.

Where is the young mathematician (statistician?) who can give us an equally comprehensive analysis of the number of photons  per cubic nm that we must elicit and detectin order to “resolve” two closely spaced point objects? And why do we always assume that these two objects have the same intensity when this is so seldom the case? What visibility criteria would be appropriate to model the problem of seeing the Earth from Alpha Centauri?

Please someone take up this quest.

Jim Pawley

James and Christine Pawley, 5446 Burley Place, Box 2348, Sechelt BC, Canada, V0N3A0 [hidden email]<mailto:[hidden email]>, Phone [phone], cell 1-604-989-6146



On Feb 13, 2017, at 7:11 AM, Alberto Diaspro <[hidden email]<mailto:[hidden email]>> wrote:

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Dear
following a discussion with the MRT staff and Wiley, they informed me that the paper is now available for free for one month. Hope this can help.
Please, have a check and let me know if it works at http://onlinelibrary.wiley.com/doi/10.1002/jemt.22834/full

Best
Alby

[Craig Brideau]

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Doesn't an infinitely sharp edge require infinite frequency content?
Therefore infinite data?

Craig

On Mon, Feb 13, 2017 at 6:57 PM, JAMES B PAWLEY <[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,
>
> Those attending the sadly-gone UBC 3D-course may remember that our first
> slogan was, "If it isn’t diffraction, it’s statistics.”
>
> Colin’s wonderful, inclusive paper does indeed wrap up the diffraction
> part superbly.
>
> But how many photons/nm*3 would you have to collect in order to obtain
> data that would allow you to fill in those beautifully narrow lines?
> Millions? (if the line width is say 1% of its “value” surely you need
> 10,000 detected photons to determine it  And how big must a pixel be to
> trace out these lovely curves without them looking blocky? Given that
> biologists habitually work with specimens capable of producing at most tens
> of detectable photons/pixel, surely it is the statistics that places the
> most severe spatial limits on the data we finally obtain.
>
> Where is the young mathematician (statistician?) who can give us an
> equally comprehensive analysis of the number of photons  per cubic nm that
> we must elicit and detectin order to “resolve” two closely spaced point
> objects? And why do we always assume that these two objects have the same
> intensity when this is so seldom the case? What visibility criteria would
> be appropriate to model the problem of seeing the Earth from Alpha Centauri?
>
> Please someone take up this quest.
>
> Jim Pawley
>
> James and Christine Pawley, 5446 Burley Place, Box 2348, Sechelt BC,
> Canada, V0N3A0 [hidden email]<mailto:[hidden email]>, Phone
> 1-604-885-0840, cell 1-604-989-6146
>
>
>
> On Feb 13, 2017, at 7:11 AM, Alberto Diaspro <[hidden email]<mailto:
> [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
> following a discussion with the MRT staff and Wiley, they informed me that
> the paper is now available for free for one month. Hope this can help.
> Please, have a check and let me know if it works at
> http://onlinelibrary.wiley.com/doi/10.1002/jemt.22834/full
>
> Best
> Alby
>
>

[Sripad Ram-2]

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Hi Jim,
Great question!! Well about 10 years ago we attempted to answer the very
same question that you ask, i.e. how many photon do you need to resolve two
closely spaced point sources. Stated differently, how does photon
statistics impact 2D/3D resolution of optical microscopes?

Here are some references to that work:

1. Beyond Rayleigh’s criterion: A resolution measure with application to
single-molecule microscopy, PNAS, 2006, 103, 4457-4462

2. A comparative study of high resolution microscopy imaging modalities
using a three-dimensional resolution measure, Optics Express,
2009,17:24377-24402

3. A resolution measure for three-dimensional microscopy, Optics
Communications, 2009, 282:1751-1761.

All of these papers should be on pubmed central as they were part of an NIH
funded study and the final accepted manuscripts were uploaded sometime in
2012.

Hope this helps.

Sripad


On Mon, Feb 13, 2017 at 6:01 PM, Craig Brideau <[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.
> *****
>
> Doesn't an infinitely sharp edge require infinite frequency content?
> Therefore infinite data?
>
> Craig
>
> On Mon, Feb 13, 2017 at 6:57 PM, JAMES B PAWLEY <[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,
> >
> > Those attending the sadly-gone UBC 3D-course may remember that our first
> > slogan was, "If it isn’t diffraction, it’s statistics.”
> >
> > Colin’s wonderful, inclusive paper does indeed wrap up the diffraction
> > part superbly.
> >
> > But how many photons/nm*3 would you have to collect in order to obtain
> > data that would allow you to fill in those beautifully narrow lines?
> > Millions? (if the line width is say 1% of its “value” surely you need
> > 10,000 detected photons to determine it  And how big must a pixel be to
> > trace out these lovely curves without them looking blocky? Given that
> > biologists habitually work with specimens capable of producing at most
> tens
> > of detectable photons/pixel, surely it is the statistics that places the
> > most severe spatial limits on the data we finally obtain.
> >
> > Where is the young mathematician (statistician?) who can give us an
> > equally comprehensive analysis of the number of photons  per cubic nm
> that
> > we must elicit and detectin order to “resolve” two closely spaced point
> > objects? And why do we always assume that these two objects have the same
> > intensity when this is so seldom the case? What visibility criteria would
> > be appropriate to model the problem of seeing the Earth from Alpha
> Centauri?
> >
> > Please someone take up this quest.
> >
> > Jim Pawley
> >
> > James and Christine Pawley, 5446 Burley Place, Box 2348, Sechelt BC,
> > Canada, V0N3A0 [hidden email]<mailto:[hidden email]>, Phone
> > 1-604-885-0840, cell 1-604-989-6146
> >
> >
> >
> > On Feb 13, 2017, at 7:11 AM, Alberto Diaspro <[hidden email]<mailto:
> > [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
> > following a discussion with the MRT staff and Wiley, they informed me
> that
> > the paper is now available for free for one month. Hope this can help.
> > Please, have a check and let me know if it works at
> > http://onlinelibrary.wiley.com/doi/10.1002/jemt.22834/full
> >
> > Best
> > Alby
> >
> >
>

[Zdenek Svindrych-2]

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*****

Hi all,
there is also a concept of "Stochastic Transfer Function" that may be able
to answer that question.

See e.g. here:

https://doi.org/10.1364/JOSAA.26.001630


It's from 2009, but it seems nobody followed that idea further... It would
be nice if someone compared various SIM methods ('Gustaffson' SIM, re-scan,
OPRA, image-scanning microscopy (airyscan), etc...) in terms of photon-
limited performance.

zdenek



---------- Původní zpráva ----------
Od: S Ram <[hidden email]>
Komu: [hidden email]
Datum: 13. 2. 2017 23:42:08
Předmět: Re: Sheppard's paper on Resolution - wall removed

"*****
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 Jim,
Great question!! Well about 10 years ago we attempted to answer the very
same question that you ask, i.e. how many photon do you need to resolve two
closely spaced point sources. Stated differently, how does photon
statistics impact 2D/3D resolution of optical microscopes?

Here are some references to that work:

1. Beyond Rayleigh’s criterion: A resolution measure with application to
single-molecule microscopy, PNAS, 2006, 103, 4457-4462

2. A comparative study of high resolution microscopy imaging modalities
using a three-dimensional resolution measure, Optics Express,
2009,17:24377-24402

3. A resolution measure for three-dimensional microscopy, Optics
Communications, 2009, 282:1751-1761.

All of these papers should be on pubmed central as they were part of an NIH
funded study and the final accepted manuscripts were uploaded sometime in
2012.

Hope this helps.

Sripad


On Mon, Feb 13, 2017 at 6:01 PM, Craig Brideau <[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.
> *****
>
> Doesn't an infinitely sharp edge require infinite frequency content?
> Therefore infinite data?
>
> Craig
>
> On Mon, Feb 13, 2017 at 6:57 PM, JAMES B PAWLEY <[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,
> >
> > Those attending the sadly-gone UBC 3D-course may remember that our first

> > slogan was, "If it isn’t diffraction, it’s statistics.”
> >
> > Colin’s wonderful, inclusive paper does indeed wrap up the diffraction
> > part superbly.
> >
> > But how many photons/nm*3 would you have to collect in order to obtain
> > data that would allow you to fill in those beautifully narrow lines?
> > Millions? (if the line width is say 1% of its “value” surely you need
> > 10,000 detected photons to determine it And how big must a pixel be to
> > trace out these lovely curves without them looking blocky? Given that
> > biologists habitually work with specimens capable of producing at most
> tens
> > of detectable photons/pixel, surely it is the statistics that places the

> > most severe spatial limits on the data we finally obtain.
> >
> > Where is the young mathematician (statistician?) who can give us an
> > equally comprehensive analysis of the number of photons per cubic nm
> that
> > we must elicit and detectin order to “resolve” two closely spaced point
> > objects? And why do we always assume that these two objects have the
same
> > intensity when this is so seldom the case? What visibility criteria
would

> > be appropriate to model the problem of seeing the Earth from Alpha
> Centauri?
> >
> > Please someone take up this quest.
> >
> > Jim Pawley
> >
> > James and Christine Pawley, 5446 Burley Place, Box 2348, Sechelt BC,
> > Canada, V0N3A0 [hidden email]<mailto:[hidden email]>, Phone
> > 1-604-885-0840, cell 1-604-989-6146
> >
> >
> >
> > On Feb 13, 2017, at 7:11 AM, Alberto Diaspro <[hidden email]<mailto:
> > [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
> > following a discussion with the MRT staff and Wiley, they informed me
> that
> > the paper is now available for free for one month. Hope this can help.
> > Please, have a check and let me know if it works at
> > http://onlinelibrary.wiley.com/doi/10.1002/jemt.22834/full 
> >
> > Best
> > Alby
> >
> >
>

"