Re: congratulations to Eric, Stefan and W.E. for Nobel Prize in Chemistry, for the development of super-resolved fluorescence microscopy

[Boehm, Ulrike-2]

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What an amazing honour for our community!

Congratulations to Stefan W. Hell, Eric Betzig and William E. Moerner!

And what next?

As Stefan said today: Back to business, because the Nobel Prize is not the end...

Good luck with all your current and future projects!

- Ulrike

________________________________________
From: Confocal Microscopy List [[hidden email]] on behalf of Sam Lord [[hidden email]]
Sent: Thursday, October 09, 2014 12:09 AM
To: [hidden email]
Subject: Re: congratulations to Eric, Stefan and W .E. for Nob el Prize in Chemistry, “ for the  development of super-reso lved fluores cence mi croscopyâ€�

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On Wed, 8 Oct 2014 09:10:30 -0500, Martin Wessendorf <[hidden email]>
wrote:
>. --According to him, the rationale for the Prize was that STED and PALM
>can be used to view biochemistry happening in living organisms.

If you read the full scientific background document, it really seems like they were
giving the award for the work done back in the 1990s (e.g. Betzig's near-field
super-resolution imaging and his PALM precursor paper in 1995). Not just his 2006
paper.

http://www.nobelprize.org/nobel_prizes/chemistry/laureates/2014/advanced-
chemistryprize2014.pdf

But it's too bad they had to limit the prize to 3. The simultaneous
PALM/STORM/FPALM papers by Betzig, Zhuang, and Sam Hess really helped ignite
the field. And the list of single-molecule folks is really long (Yanagida, Webb, Zare,
Vale, Orrit, Rigler, Xie, Cremer…). That said, Hell, Betzig, and Moerner are
unquestionably deserving and it's really exciting. (W.E. was my PhD advisor.)

I wrote a blog post about it here: http://blog.everydayscientist.com/?p=3254

-Sam

[Andrew York]

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I think the list will find this article entertaining:
http://www.slate.com/articles/health_and_science/science/2014/10/chemistry_nobel_prize_winners_see_microscope_images_they_made_possible_photos.html

Pay particular attention to the images.

Unrelated: I assume Betzig's award is primarily due to his demonstration of
PALM, and related follow-up work. I assume Hell's is similarly for his
demonstration of STED. What work of Moerner's are they thinking of? Is
there one specific paper/idea, like with Betzig and Hell, or is it for
decades of work on single molecules in general?
On Oct 10, 2014 4:25 AM, "Andreas Schönle" <
[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 Guy, Nuno, dear list members!
>
> Surely this discussion will now go on for a while but I would like to set a
> few facts straight, so it can be put on solid ground.
>
> 1)
> Stefan Hell was never a student of Chirstoph Cremer. His PhD advisor was
> Prof. S. Hunklinger and both 4pi and STED microscopy were conceived by him
> independently. The principle of STED is completely unrelated to 4pi and 4pi
> has nothing to do with what the Nobel prize was awarded for (see below).
> 2)
> 4pi microscopy is not a super-resolution method (it is diffraction
> limited).
> Christoph Cremer's original proposal to use light coming from all
> directions
> was aimed at increasing lateral resolution and was based on the wrong
> assumption that inverting the light-field of a point source <<lambda by
> means of a hologram could beat the diffraction limit.
> (this neglects the fact that the near field decays exponentially and that
> this cannot be inverted in conventional materials)
> 3)
> Localization is not resolution. Impressive work was done localizing single
> or several light sources separable by spectral properties, stepwise
> bleaching etc. - usually with the goal of measuring distances. But dense
> images can (as of now) only be obtained by time-sequentially switching
> molecules on and off.
> This was first realized for ensembles in STED and molecule-by-molecule in
> PALM/STORM/FPALM.
>
> Again, this is not to diminish anybody's work in the field. Over the years
> I
> have read many articles describing important and inspiring work. And I
> wholeheartedly agree that the field would not be where it is without these
> contributions.
> But when discussing who should be honored for finding the key to circumvent
> the diffraction limit in optical imaging, it is important to identify those
> people that had the right idea, recognized its importance and proved this
> by
> actually putting it to work.
>
> Best regards,
> Andreas
>
> --
> Andreas Schoenle, Dr.
> Abberior Instruments GmbH, D-37077 Goettingen, Germany
>
> phone: [phone]
> fax: [phone]
> http://www.abberior-instruments.com
> mailto:[hidden email]
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]]
> On
> Behalf Of Nuno Moreno
> Sent: Freitag, 10. Oktober 2014 00:19
> To: [hidden email]
> Subject: Re: congratulations to Eric, Stefan and W . E. for Nob el P rize
> in
> Chemistry, ³ for t he development of super-reso lved fluores cenc e mi
> croscopy²
>
> *****
> 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 Guy
>
> You're absolutely right. I actually sent an internal email (for the
> institute) a few hours after the announcement with, among other stuff,
> this:
>
> "As always there are many others that should be on the laureates list but
> one needed the final click. In my opinion Cremer from Heidelberg university
> should be one of them ...."
>
> Nuno Moreno
>
>
>
>
>
> On 09 Oct 2014, at 02:16, Guy Cox <[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.
> > *****
> >
> > The three names is always a problem - especially when the prize is
> awarded
> essentially for two different techniques.  Christoph Cremer (Stefan's
> supervisor) could well also feel he'd been passed over.
> >
> >                                                            Guy
> >
>

[Guy Cox-2]

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Can I please make it clear that neither I, nor Nuno, nor other contributors who have mentioned other people, grudge these awards in the slightest.  The three winners (and I have known Stefan Hell for 30 years) all totally deserve their awards.  What's more, it is a boost for all of us that super-resolution microscopy has made the Nobel.  But as with many other cases in the past, the strait-jacket of only 3 winners (max) means that other deserving people miss out.  This is particularly the case where two topics share the same prize.  (cf EM and STM some years ago).  So, while congratulating the three winners, it is good that we recall other contributors, so that they will know that even if they didn't get the prize, they are still remembered by the community.  

                                                  Guy

Guy Cox, Honorary Associate Professor
School of Medical Sciences

Australian Centre for Microscopy and Microanalysis,
Madsen, F09, University of Sydney, NSW 2006

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Andreas Schönle
Sent: Friday, 10 October 2014 7:25 PM
To: [hidden email]
Subject: Re: congratulations to Eric, Stefan and W.E. for Nobel Prize in Chemistry, for the development of super-resolved fluorescence microscopy

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Dear Guy, Nuno, dear list members!

Surely this discussion will now go on for a while but I would like to set a few facts straight, so it can be put on solid ground.

1)
Stefan Hell was never a student of Chirstoph Cremer. His PhD advisor was Prof. S. Hunklinger and both 4pi and STED microscopy were conceived by him independently. The principle of STED is completely unrelated to 4pi and 4pi has nothing to do with what the Nobel prize was awarded for (see below).
2)
4pi microscopy is not a super-resolution method (it is diffraction limited).
Christoph Cremer's original proposal to use light coming from all directions was aimed at increasing lateral resolution and was based on the wrong assumption that inverting the light-field of a point source <<lambda by means of a hologram could beat the diffraction limit.
(this neglects the fact that the near field decays exponentially and that this cannot be inverted in conventional materials)
3)
Localization is not resolution. Impressive work was done localizing single or several light sources separable by spectral properties, stepwise bleaching etc. - usually with the goal of measuring distances. But dense images can (as of now) only be obtained by time-sequentially switching molecules on and off.
This was first realized for ensembles in STED and molecule-by-molecule in PALM/STORM/FPALM.

Again, this is not to diminish anybody's work in the field. Over the years I have read many articles describing important and inspiring work. And I wholeheartedly agree that the field would not be where it is without these contributions.
But when discussing who should be honored for finding the key to circumvent the diffraction limit in optical imaging, it is important to identify those people that had the right idea, recognized its importance and proved this by actually putting it to work.

Best regards,
Andreas

--
Andreas Schoenle, Dr.
Abberior Instruments GmbH, D-37077 Goettingen, Germany
 
phone: [phone]
fax: [phone]
http://www.abberior-instruments.com
mailto:[hidden email]

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Nuno Moreno
Sent: Freitag, 10. Oktober 2014 00:19
To: [hidden email]
Subject: Re: congratulations to Eric, Stefan and W . E. for Nob el P rize in Chemistry, ³ for t he development of super-reso lved fluores cenc e mi croscopy²

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Dear Guy

You're absolutely right. I actually sent an internal email (for the
institute) a few hours after the announcement with, among other stuff, this:

"As always there are many others that should be on the laureates list but one needed the final click. In my opinion Cremer from Heidelberg university should be one of them ...."

Nuno Moreno





On 09 Oct 2014, at 02:16, Guy Cox <[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.
> *****
>
> The three names is always a problem - especially when the prize is
> awarded
essentially for two different techniques.  Christoph Cremer (Stefan's
supervisor) could well also feel he'd been passed over.
>
>                                                            Guy
>

[Andreas Bruckbauer]

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 In the 'Advanced Information' from the Nobel Committee they actually put the publications in bold which were key for the award, in the case of WE Moerner these are:

Moerner WE and Kador L (1989) Optical detection and spectroscopy of single molecules in a solid. Phys. Rev. Lett.62:2535-2538.

Dickson RM, Cubitt AB, Tsien RY and Moerner WE (1997) On/off blinking and switching behaviour of single molecules of green fluorescent protein. Nature 388:355-358.
 

 So it is quite clear, the first detection of single molecules in a solid and the blinking behaviour of GFP, which links it quite nicely to Eric Betzig's work. In Betzig's case the early NSMO work is not in bold, but the 'Proposed Method' paper from 1995 is, putting him ahead of others in the field. Sometimes it is good to publish an idea on its own.

Anyhow, i think with the effort of the whole community including funding organisations, journal editors and countless students, this Nobel prize would not have been possible, well done everyone! Let's not forget, there were only a few microscope techniques which were worth a nobel prize before: Zsigmondi (ultramicroscope), Zernike (phase contrast), Ruska (Electronmicroscope, Binning & Rohrer (scanning tunnelling microscope).

best wishes

Andreas


 

-----Original Message-----
From: Andrew York <[hidden email]>
To: CONFOCALMICROSCOPY <[hidden email]>
Sent: Fri, 10 Oct 2014 13:19
Subject: Re: congratulations to Eric, Stefan and W.E. for Nobel Prize in Chemistry, for the development of super-resolved fluorescence microscopy


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I think the list will find this article entertaining:
http://www.slate.com/articles/health_and_science/science/2014/10/chemistry_nobel_prize_winners_see_microscope_images_they_made_possible_photos.html

Pay particular attention to the images.

Unrelated: I assume Betzig's award is primarily due to his demonstration of
PALM, and related follow-up work. I assume Hell's is similarly for his
demonstration of STED. What work of Moerner's are they thinking of? Is
there one specific paper/idea, like with Betzig and Hell, or is it for
decades of work on single molecules in general?
On Oct 10, 2014 4:25 AM, "Andreas Schönle" <
[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 Guy, Nuno, dear list members!
>
> Surely this discussion will now go on for a while but I would like to set a
> few facts straight, so it can be put on solid ground.
>
> 1)
> Stefan Hell was never a student of Chirstoph Cremer. His PhD advisor was
> Prof. S. Hunklinger and both 4pi and STED microscopy were conceived by him
> independently. The principle of STED is completely unrelated to 4pi and 4pi
> has nothing to do with what the Nobel prize was awarded for (see below).
> 2)
> 4pi microscopy is not a super-resolution method (it is diffraction
> limited).
> Christoph Cremer's original proposal to use light coming from all
> directions
> was aimed at increasing lateral resolution and was based on the wrong
> assumption that inverting the light-field of a point source <<lambda by
> means of a hologram could beat the diffraction limit.
> (this neglects the fact that the near field decays exponentially and that
> this cannot be inverted in conventional materials)
> 3)
> Localization is not resolution. Impressive work was done localizing single
> or several light sources separable by spectral properties, stepwise
> bleaching etc. - usually with the goal of measuring distances. But dense
> images can (as of now) only be obtained by time-sequentially switching
> molecules on and off.
> This was first realized for ensembles in STED and molecule-by-molecule in
> PALM/STORM/FPALM.
>
> Again, this is not to diminish anybody's work in the field. Over the years
> I
> have read many articles describing important and inspiring work. And I
> wholeheartedly agree that the field would not be where it is without these
> contributions.
> But when discussing who should be honored for finding the key to circumvent
> the diffraction limit in optical imaging, it is important to identify those
> people that had the right idea, recognized its importance and proved this
> by
> actually putting it to work.
>
> Best regards,
> Andreas
>
> --
> Andreas Schoenle, Dr.
> Abberior Instruments GmbH, D-37077 Goettingen, Germany
>
> phone: [phone]
> fax: [phone]
> http://www.abberior-instruments.com
> mailto:[hidden email]
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]]
> On
> Behalf Of Nuno Moreno
> Sent: Freitag, 10. Oktober 2014 00:19
> To: [hidden email]
> Subject: Re: congratulations to Eric, Stefan and W . E. for Nob el P rize
> in
> Chemistry, ³ for t he development of super-reso lved fluores cenc e mi
> croscopy²
>
> *****
> 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 Guy
>
> You're absolutely right. I actually sent an internal email (for the
> institute) a few hours after the announcement with, among other stuff,
> this:
>
> "As always there are many others that should be on the laureates list but
> one needed the final click. In my opinion Cremer from Heidelberg university
> should be one of them ...."
>
> Nuno Moreno
>
>
>
>
>
> On 09 Oct 2014, at 02:16, Guy Cox <[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.
> > *****
> >
> > The three names is always a problem - especially when the prize is
> awarded
> essentially for two different techniques.  Christoph Cremer (Stefan's
> supervisor) could well also feel he'd been passed over.
> >
> >                                                            Guy
> >
>

 

[Steve Baer]

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Dear List members,

I would like to respond to Andreas Schoenle’s post. Even if Christoph Cremer was not Hell’s thesis advisor, he was a professor in Hell’s department during most of Hell's years as a graduate student, and their common interest in superresolution microscopy was an important connection. Furthermore, two decades earlier, Cremer and his brother had filed a patent application on the basic concept of the 4Pi microscope, which was to illuminate a point coherently from all directions, rather from just the one side as in previous microscopes, and then to scan that illuminated point in a raster pattern, detect the light emitted from the scanned point, and form an image with the detected light. In their patent application, the Cremer brothers even named their instrument the 4Pi microscope. That application is available for download from the German Patent Office.

Cremer has been gracious enough to credit Hell with improving his original concept by combining it with the confocal microscope concept, though needless to say, a patent examiner might call such a combination obvious. Hell’s ’09 Nature Methods review article, “Microscopy and its focal switch”, included a discussion of the history of the 4Pi microscope. By the time he wrote that article, Hell was obviously aware that the Cremer brothers were the original inventors of that instrument. Yet of the four references on the 4Pi microscope that Hell cited in that paper, the earliest was his own 1990 patent application, implying that Hell’s was the original inventor of that instrument. In that paper, Hell’s only acknowledgement of the Cremer’s work was, “based on inadequate assumptions about the nature of diffraction and hence did not lead to valid conclusions.” This comment speaks for itself.

Andreas is correct that this year’s Chemistry Nobel prize was awarded for the breakthrough in resolution, enabling not just an improvement over the Abbe diffraction limit, such as the 40% improvement afforded by Minsky's confocal microscope, but for the complete elimination of that limit, in principal enabling resolution at molecular dimensions. That breakthrough improvement is embodied in the “STED resolution equation,”

d = λ/2n sin α √ (1 + a Imax/Is)
which shows that as the intensity Imax of the STED beam increases without limit, the minimum distance d between just-resolvable points goes to zero without limit.

Hell claims that the first microscope embodying that breakthrough resolution performance was described in his 1994 paper with Wichmann. That is simply untrue, as anyone can easily see looking at the paper. The microscope described in the Hell and Wichmann paper is limited to a resolution improvement of about fourfold. At its maximum resolution, the fluorescent image is quenched out of existence, so further increases in the intensity of the STED beam make no difference. I urge members of this discussion group to download the 1994 Hell and Wichmann paper from Hell’s web site, and confirm this for themselves.

The key innovation that allowed a STED microscope to break free of the diffraction limit was to give the STED beam a “zero-point,” which is to say a point of zero intensity surrounded by points of non-zero intensity, so that at any distance from the zero-point, however close, the intensity is greater than zero, and as such can be increased to an arbitrary level by raising the mean intensity of the STED beam. Even at such a high mean intensity, the central point of the STED beam will remain at zero intensity, so the central point of the fluorescent spot remains at its initial level, regardless of how much the spot’s width is reduced.

The first STED microscope to contain the requisite zero-point, thus completely transcending the diffraction limit, was the STED microscope I invented independently of Hell and Wichmann and described in the patent application for US Pat. 5,866,911, filed the same year they submitted their original paper. I am extremely grateful to Guy for including a reference to that patent in his new book, and to Christoph Cremer, Barry Masters, Rainer Heintzmann, and the late Mats Gustafsson for citing that that patent in review papers.

Steve Baer

On Oct 10, 2014, at 4:24 AM, Andreas Schönle <[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 Guy, Nuno, dear list members!
>
> Surely this discussion will now go on for a while but I would like to set a
> few facts straight, so it can be put on solid ground.
>
> 1)
> Stefan Hell was never a student of Chirstoph Cremer. His PhD advisor was
> Prof. S. Hunklinger and both 4pi and STED microscopy were conceived by him
> independently. The principle of STED is completely unrelated to 4pi and 4pi
> has nothing to do with what the Nobel prize was awarded for (see below).
> 2)
> 4pi microscopy is not a super-resolution method (it is diffraction limited).
> Christoph Cremer's original proposal to use light coming from all directions
> was aimed at increasing lateral resolution and was based on the wrong
> assumption that inverting the light-field of a point source <<lambda by
> means of a hologram could beat the diffraction limit.
> (this neglects the fact that the near field decays exponentially and that
> this cannot be inverted in conventional materials)
> 3)
> Localization is not resolution. Impressive work was done localizing single
> or several light sources separable by spectral properties, stepwise
> bleaching etc. - usually with the goal of measuring distances. But dense
> images can (as of now) only be obtained by time-sequentially switching
> molecules on and off.
> This was first realized for ensembles in STED and molecule-by-molecule in
> PALM/STORM/FPALM.
>
> Again, this is not to diminish anybody's work in the field. Over the years I
> have read many articles describing important and inspiring work. And I
> wholeheartedly agree that the field would not be where it is without these
> contributions.
> But when discussing who should be honored for finding the key to circumvent
> the diffraction limit in optical imaging, it is important to identify those
> people that had the right idea, recognized its importance and proved this by
> actually putting it to work.
>
> Best regards,
> Andreas
>
> --
> Andreas Schoenle, Dr.
> Abberior Instruments GmbH, D-37077 Goettingen, Germany
>  
> phone: [phone]
> fax: [phone]
> http://www.abberior-instruments.com
> mailto:[hidden email]
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]] On
> Behalf Of Nuno Moreno
> Sent: Freitag, 10. Oktober 2014 00:19
> To: [hidden email]
> Subject: Re: congratulations to Eric, Stefan and W . E. for Nob el P rize in
> Chemistry, ³ for t he development of super-reso lved fluores cenc e mi
> croscopy²
>
> *****
> 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 Guy
>
> You're absolutely right. I actually sent an internal email (for the
> institute) a few hours after the announcement with, among other stuff, this:
>
> "As always there are many others that should be on the laureates list but
> one needed the final click. In my opinion Cremer from Heidelberg university
> should be one of them ...."
>
> Nuno Moreno
>
>
>
>
>
> On 09 Oct 2014, at 02:16, Guy Cox <[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.
>> *****
>>
>> The three names is always a problem - especially when the prize is awarded
> essentially for two different techniques.  Christoph Cremer (Stefan's
> supervisor) could well also feel he'd been passed over.
>>
>>                                                           Guy
>>

[Antonio Jose Pereira]

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Hi,

These discussions could go on forever and we can gain with that,
for we will pick back the historical sequences and learn ever more on the subject.

We all understand what it is to have had a role in the development of something and then suffering from a nonlinear curve of recognition.

Answering to Steve, in my opinion the key creative spark is indeed to make use of nonlinearity of light-matter interaction. That was evidently there, in 94 Hell's paper. The factor of 4.5 appears there just a technical limitation for using STED beams which don't have a zero finitely close to the maximum.

They don't put the factor 4.5 as one puts the factor 2 in non-saturated structured illumination, for example. They seem, in 94 Hell's paper, to be very aware that the principle could lead the resolution infinitely far, i.e., that it is not about the wavelength of the photons anymore, it is about ... well deBroglie wavelengths in the sample, if you wish.

That said, the community is very aware that these breakthroughs are always much more than 3 people ...

Antonio Pereira
IBMC - Porto
 



António Pereira
IBMC - Instituto de Biologia Molecular e Celular
Chromosome Instability and Dynamics Group

Rua do Campo Alegre, 823
4150-180 Porto
Portugal

[phone] (ext# 1620)
[hidden email]

-----Confocal Microscopy List <[hidden email]> wrote: -----
To: [hidden email]
From: Steve Baer
Sent by: Confocal Microscopy List
Date: 10/10/2014 03:57PM
Subject: Re: congratulations to Eric, Stefan and W.E. for Nobel Prize in Chemistry, for the development of super-resolved fluorescence microscopy

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Dear List members,

I would like to respond to Andreas Schoenle’s post. Even if Christoph Cremer was not Hell’s thesis advisor, he was a professor in Hell’s department during most of Hell's years as a graduate student, and their common interest in superresolution microscopy was an important connection. Furthermore, two decades earlier, Cremer and his brother had filed a patent application on the basic concept of the 4Pi microscope, which was to illuminate a point coherently from all directions, rather from just the one side as in previous microscopes, and then to scan that illuminated point in a raster pattern, detect the light emitted from the scanned point, and form an image with the detected light. In their patent application, the Cremer brothers even named their instrument the 4Pi microscope. That application is available for download from the German Patent Office.

Cremer has been gracious enough to credit Hell with improving his original concept by combining it with the confocal microscope concept, though needless to say, a patent examiner might call such a combination obvious. Hell’s ’09 Nature Methods review article, “Microscopy and its focal switch”, included a discussion of the history of the 4Pi microscope. By the time he wrote that article, Hell was obviously aware that the Cremer brothers were the original inventors of that instrument. Yet of the four references on the 4Pi microscope that Hell cited in that paper, the earliest was his own 1990 patent application, implying that Hell’s was the original inventor of that instrument. In that paper, Hell’s only acknowledgement of the Cremer’s work was, “based on inadequate assumptions about the nature of diffraction and hence did not lead to valid conclusions.” This comment speaks for itself.

Andreas is correct that this year’s Chemistry Nobel prize was awarded for the breakthrough in resolution, enabling not just an improvement over the Abbe diffraction limit, such as the 40% improvement afforded by Minsky's confocal microscope, but for the complete elimination of that limit, in principal enabling resolution at molecular dimensions. That breakthrough improvement is embodied in the “STED resolution equation,”

d = λ/2n sin α √ (1 + a Imax/Is)
which shows that as the intensity Imax of the STED beam increases without limit, the minimum distance d between just-resolvable points goes to zero without limit.

Hell claims that the first microscope embodying that breakthrough resolution performance was described in his 1994 paper with Wichmann. That is simply untrue, as anyone can easily see looking at the paper. The microscope described in the Hell and Wichmann paper is limited to a resolution improvement of about fourfold. At its maximum resolution, the fluorescent image is quenched out of existence, so further increases in the intensity of the STED beam make no difference. I urge members of this discussion group to download the 1994 Hell and Wichmann paper from Hell’s web site, and confirm this for themselves.

The key innovation that allowed a STED microscope to break free of the diffraction limit was to give the STED beam a “zero-point,” which is to say a point of zero intensity surrounded by points of non-zero intensity, so that at any distance from the zero-point, however close, the intensity is greater than zero, and as such can be increased to an arbitrary level by raising the mean intensity of the STED beam. Even at such a high mean intensity, the central point of the STED beam will remain at zero intensity, so the central point of the fluorescent spot remains at its initial level, regardless of how much the spot’s width is reduced.

The first STED microscope to contain the requisite zero-point, thus completely transcending the diffraction limit, was the STED microscope I invented independently of Hell and Wichmann and described in the patent application for US Pat. 5,866,911, filed the same year they submitted their original paper. I am extremely grateful to Guy for including a reference to that patent in his new book, and to Christoph Cremer, Barry Masters, Rainer Heintzmann, and the late Mats Gustafsson for citing that that patent in review papers.

Steve Baer

On Oct 10, 2014, at 4:24 AM, Andreas Schönle <[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 Guy, Nuno, dear list members!
>
> Surely this discussion will now go on for a while but I would like to set a
> few facts straight, so it can be put on solid ground.
>
> 1)
> Stefan Hell was never a student of Chirstoph Cremer. His PhD advisor was
> Prof. S. Hunklinger and both 4pi and STED microscopy were conceived by him
> independently. The principle of STED is completely unrelated to 4pi and 4pi
> has nothing to do with what the Nobel prize was awarded for (see below).
> 2)
> 4pi microscopy is not a super-resolution method (it is diffraction limited).
> Christoph Cremer's original proposal to use light coming from all directions
> was aimed at increasing lateral resolution and was based on the wrong
> assumption that inverting the light-field of a point source <<lambda by
> means of a hologram could beat the diffraction limit.
> (this neglects the fact that the near field decays exponentially and that
> this cannot be inverted in conventional materials)
> 3)
> Localization is not resolution. Impressive work was done localizing single
> or several light sources separable by spectral properties, stepwise
> bleaching etc. - usually with the goal of measuring distances. But dense
> images can (as of now) only be obtained by time-sequentially switching
> molecules on and off.
> This was first realized for ensembles in STED and molecule-by-molecule in
> PALM/STORM/FPALM.
>
> Again, this is not to diminish anybody's work in the field. Over the years I
> have read many articles describing important and inspiring work. And I
> wholeheartedly agree that the field would not be where it is without these
> contributions.
> But when discussing who should be honored for finding the key to circumvent
> the diffraction limit in optical imaging, it is important to identify those
> people that had the right idea, recognized its importance and proved this by
> actually putting it to work.
>
> Best regards,
> Andreas
>
> --
> Andreas Schoenle, Dr.
> Abberior Instruments GmbH, D-37077 Goettingen, Germany
>  
> phone: [phone]
> fax: [phone]
> http://www.abberior-instruments.com
> mailto:[hidden email] 
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]] On
> Behalf Of Nuno Moreno
> Sent: Freitag, 10. Oktober 2014 00:19
> To: [hidden email]
> Subject: Re: congratulations to Eric, Stefan and W . E. for Nob el P rize in
> Chemistry, ³ for t he development of super-reso lved fluores cenc e mi
> croscopy²
>
> *****
> 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 Guy
>
> You're absolutely right. I actually sent an internal email (for the
> institute) a few hours after the announcement with, among other stuff, this:
>
> "As always there are many others that should be on the laureates list but
> one needed the final click. In my opinion Cremer from Heidelberg university
> should be one of them ...."
>
> Nuno Moreno
>
>
>
>
>
> On 09 Oct 2014, at 02:16, Guy Cox <[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.
>> *****
>>
>> The three names is always a problem - especially when the prize is awarded
> essentially for two different techniques.  Christoph Cremer (Stefan's
> supervisor) could well also feel he'd been passed over.
>>
>>                                                           Guy
>>

[Peng Xi-2]

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*****

I fully agree with Prof. Guy Cox. We sincerely congratulate all the three
winners, because we all know that they well deserve it. And, this will
bring all of us a brighter future, because microscopy gets back to people's
vision. However, if being interpreted wrongly, the only-3-winner policy may
become a source of internal conflict, which hurts the community at large.
Let me share an old Chinese historic story: "Two peaches kill three
warriors".
----
 It was 2000BC in ancient China, there are three warriors who are great
generals during the war time. In peace time however, they respect nobody
because of their great contributions to the kingdom. So the king was
worried, and he consulted his senate. The senate said: "You can award them
with two best peaches of our country." So the king followed his suggestion,
saying that he will give the award to those with greatest contribution.

So the three warriers were invited to the ceremony. The first warrior
Gongsun, a very quick tempered person, responded:"I had killed one giant
tiger to save the king's life, I should deserve one." So he took one peach.
The second warrior Gu said:"A true warrior shows his power at war. I have
killed many enemies and won many hard battles, I am sure to have the right
of winning a peach as award." So he also took one.

The third warrior Tian was surprised at this fact. He didn't speak earlier
because he want to show some respect to his fellows, but the result is very
unacceptable to him. So he said, " You mentioned killing tiger and killing
people. Think about my deeds. Once our king is traveling across the Yellow
River, then a dragon jumps out of water and swallowed the lovely son of the
king. I fought with the monster for several hours, slaughter its head, and
saved the princess. Yet, I do not have any award. Enjoy the peach,
gentlemen!" After saying this, he took out his sward, and suicided.

Seeing the blood of Tian, Gongsun started to think: "Surely my contribution
is great, but comparing with Tian, it is nothing. Yet I see only the award,
and ignores my friends, this simply reflects my blindness and selfishness
in my character, which is completely unacceptable for a warrior." Thinking
that this kills his best friend, he took out his sward and cut his belly
also.
Then Gu cried: "We three had fought so many battles together, and already
swored to be brothers of different family name. We sowred to live and to
die together. Yet now, I flatter myself to compete with your contributions,
this is no rightousness; I am still alive while you both are dead, this is
no faith; I dare not to kill myself, this is no courage. I see no point
that I as a warrior should live like this. " So, he ended his life as well.

The senate won his trick, with two intact peaches.
-----

I wish that this ancient wisdom may bring more unity and harmony for our
community.  Only by united can our community get the full blessings from
super-resolution microscopy wining Nobel prize.
You can check out the story by yourself with help of google translate
through:   http://baike.baidu.com/view/44773.htm         By no means should
one connect the names and award to anybody or any organization in real life
in any form.

      Peng


Sincerely,
Peng Xi
Ph. D.    Associate Professor
Dept. of Biomedical Engineering, College of Engineering
Peking University, Beijing, China
Tel: [phone]
Email: [hidden email]
http://bme.pku.edu.cn/~xipeng/

On Fri, Oct 10, 2014 at 8:31 PM, Guy Cox <[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.
> *****
>
> Can I please make it clear that neither I, nor Nuno, nor other
> contributors who have mentioned other people, grudge these awards in the
> slightest.  The three winners (and I have known Stefan Hell for 30 years)
> all totally deserve their awards.  What's more, it is a boost for all of us
> that super-resolution microscopy has made the Nobel.  But as with many
> other cases in the past, the strait-jacket of only 3 winners (max) means
> that other deserving people miss out.  This is particularly the case where
> two topics share the same prize.  (cf EM and STM some years ago).  So,
> while congratulating the three winners, it is good that we recall other
> contributors, so that they will know that even if they didn't get the
> prize, they are still remembered by the community.
>
>                                                   Guy
>
> Guy Cox, Honorary Associate Professor
> School of Medical Sciences
>
> Australian Centre for Microscopy and Microanalysis,
> Madsen, F09, University of Sydney, NSW 2006
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]]
> On Behalf Of Andreas Schönle
> Sent: Friday, 10 October 2014 7:25 PM
> To: [hidden email]
> Subject: Re: congratulations to Eric, Stefan and W.E. for Nobel Prize in
> Chemistry, for the development of super-resolved fluorescence microscopy
>
> *****
> 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 Guy, Nuno, dear list members!
>
> Surely this discussion will now go on for a while but I would like to set
> a few facts straight, so it can be put on solid ground.
>
> 1)
> Stefan Hell was never a student of Chirstoph Cremer. His PhD advisor was
> Prof. S. Hunklinger and both 4pi and STED microscopy were conceived by him
> independently. The principle of STED is completely unrelated to 4pi and 4pi
> has nothing to do with what the Nobel prize was awarded for (see below).
> 2)
> 4pi microscopy is not a super-resolution method (it is diffraction
> limited).
> Christoph Cremer's original proposal to use light coming from all
> directions was aimed at increasing lateral resolution and was based on the
> wrong assumption that inverting the light-field of a point source <<lambda
> by means of a hologram could beat the diffraction limit.
> (this neglects the fact that the near field decays exponentially and that
> this cannot be inverted in conventional materials)
> 3)
> Localization is not resolution. Impressive work was done localizing single
> or several light sources separable by spectral properties, stepwise
> bleaching etc. - usually with the goal of measuring distances. But dense
> images can (as of now) only be obtained by time-sequentially switching
> molecules on and off.
> This was first realized for ensembles in STED and molecule-by-molecule in
> PALM/STORM/FPALM.
>
> Again, this is not to diminish anybody's work in the field. Over the years
> I have read many articles describing important and inspiring work. And I
> wholeheartedly agree that the field would not be where it is without these
> contributions.
> But when discussing who should be honored for finding the key to
> circumvent the diffraction limit in optical imaging, it is important to
> identify those people that had the right idea, recognized its importance
> and proved this by actually putting it to work.
>
> Best regards,
> Andreas
>
> --
> Andreas Schoenle, Dr.
> Abberior Instruments GmbH, D-37077 Goettingen, Germany
>
> phone: [phone]
> fax: [phone]
> http://www.abberior-instruments.com
> mailto:[hidden email]
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]]
> On Behalf Of Nuno Moreno
> Sent: Freitag, 10. Oktober 2014 00:19
> To: [hidden email]
> Subject: Re: congratulations to Eric, Stefan and W . E. for Nob el P rize
> in Chemistry, ³ for t he development of super-reso lved fluores cenc e mi
> croscopy²
>
> *****
> 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 Guy
>
> You're absolutely right. I actually sent an internal email (for the
> institute) a few hours after the announcement with, among other stuff,
> this:
>
> "As always there are many others that should be on the laureates list but
> one needed the final click. In my opinion Cremer from Heidelberg university
> should be one of them ...."
>
> Nuno Moreno
>
>
>
>
>
> On 09 Oct 2014, at 02:16, Guy Cox <[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.
> > *****
> >
> > The three names is always a problem - especially when the prize is
> > awarded
> essentially for two different techniques.  Christoph Cremer (Stefan's
> supervisor) could well also feel he'd been passed over.
> >
> >                                                            Guy
> >
>