http://confocal-microscopy-list.275.s1.nabble.com/Inventor-of-fluorescence-Ploemopak-in-running-for-Nobel-Prize-website-on-his-early-technology-tp7580208p7580270.html
electron microscope, something that he first demonstrated in 1933. He
>
>
>-----Original Message-----
>From: Confocal Microscopy List
>[mailto:
[hidden email]] On Behalf Of Tim Feinstein
>Sent: Thursday, 25 April 2013 10:19 PM
>To:
[hidden email]
>Subject: Re: Inventor of fluorescence Ploemopak in running for Nobel
>Prize + website on his early technology
>
>*****
>To join, leave or search the confocal microscopy listserv, go to:
>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy>*****
>
>Hi guys,
>
>I think it can be argued that the 1993 PCR prize honored a technique
>rather than a specific experimental advance.
>
>/ 2c
>
>Cheers,
>
>
>TF
>
>Timothy Feinstein, Ph.D.
>Visiting Research Associate
>Laboratory for GPCR Biology
>Dept. of Pharmacology & Chemical Biology
>University of Pittsburgh, School of Medicine
>BST W1301, 200 Lothrop St.
>Pittsburgh, PA 15261
>
>On Apr 25, 2013, at 5:47 AM, Mark Cannell <
[hidden email]> wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy>> *****
>>
>> Hi Guy
>>
>> I was not trying to be unkind. Just dealing with the point that
>>the Nobel is for the experimental side -which may come from new
>>technology development. In the case of GFP, I agree that others
>>contributed but it was the demonstration of the multiple benefits
>>of the application that was probably a key factor.l. As a case in
>>point take PET -most of the engineers and mathematicians involved
>>did not get the recognition that was deserved but it was the
>>pioneering application that led to the prize. at least that's how I
>>understand it.
>>
>> Cheers
>>
>> was probably they decider in who should get the
>> On 25/04/2013, at 10:19 AM, 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>>> *****
>>>
>>> Mark,
>>>
>>> I think this is a bit unkind. Every Nobel has people who
>>>had done pioneering work who didn't get the gong. One in
>>>particular involved a close friend and colleague of mine with whom
>>>I have published several papers. Another we all know about was
>>>the GFP prize. But the rules only allow 3 people max, so there
>>>are always going to be problems of this sort.
> >>
>>> To deny the scale of Ploem's breakthrough is ridiculous,
>>>looking at where fluorescence microscopy has got to as a result of
>>>his work. My background is in Cyril Darlington's department at
>>>Oxford, where fluorescence microscopy was used from a very early
>>>date, but it was a very esoteric and extremely dangerous technique
>>>only used by very highly trained technical staff. To emphasize
>>>the dangers, when I started in Sydney I knew Professor Y-T Tchan
>>>who was then emeritus professor of microbiology. He was blind in
>>>one eye as a result of a student pulling out the barrier filter of
>>>a diascopic fluorescence microscope (when he was at the Sorbonne).
>>>
>>> Ploem made fluorescence microscopy a routine part of
>>>cell biology and this led to incredible advances which would not
>>>have been possible without his research. I do think that the
>>>pioneers of immuno-fluorescence also deserve a Nobel!
> >>
>>> Guy
>>>
>>> -----Original Message-----
>>> From: Confocal Microscopy List
>>>[mailto:
[hidden email]] On Behalf Of Mark Cannell
>>> Sent: Thursday, 25 April 2013 6:35 PM
>>> To:
[hidden email]
>>> Subject: Re: Inventor of fluorescence Ploemopak in running for
>>>Nobel Prize + website on his early technology
>>>
>>> *****
>>> To join, leave or search the confocal microscopy listserv, go to:
>>>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy>>> *****
>>>
>>> Hi John
>>>
>>> I think the Nobel is for experimental work, not just technology
>>>so I'm not sure the filter cube is a candidate. While we are
>>>talking about fluorescence microscopy what about the development
>>>and application of live cell real time fluorescence
>>>imaging/microscpectrofluorimetry? That developed some time later
>>>but I don't know whose work predated my own work on video rate Ca
>>>imaging ... Any ideas/references ?
>>>
>>> Cheers Mark
>>>
>>> On 25/04/2013, at 8:04 AM, John Oreopoulos
>>><
[hidden email]> wrote:
>>>
>>>> *****
>>>> To join, leave or search the confocal microscopy listserv, go to:
>>>>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy>>>> *****
>>>>
>>>> I have been thinking about this for over a week now off and on,
>>>>and I think I can put down in writing the main technical reasons
>>>>why Ploem's contribution IS worthy of a Nobel prize (recognizing
>>>>of course that I might be a bit biased towards seeing another
>>>>prize being awarded to imaging science and technology, of course).
>>>>
>>>> 1. Older transmitted (diascopic) fluorescence illumination
>>>>employed a condenser lens to focus illumination light in order to
>>>>excite fluorochromes / fluorescent probes embedded in a
>>>>microscopic sample. This required that the condenser lens be well
>>>>aligned to the objective lens (Kohler illumination). Many of the
>>>>early diascopic fluorescence microscopes also employed a
>>>>darkfield type of illumination to reduce the background light,
>>>>and in doing so necessitated the application of a high NA
>>>>oil-immersion condenser lens which further complicated the
>>>>optical alignment. The switch to incident (episcopic)
>>>>fluorescence illumination allowed the objective lens to also take
>>>>on the role of the condenser lens (concentrating/focusing the
>>>>illumination light into the centre of the field of view and onto
>>>>the correct focal plane), and since the excitation and emission
>>>>light path traversed the same lens, the optical required
>>>>alignment was achieved automatically.
>>>>
>>>> 2. Related to point 1, with the objective now playing the role
>>>>of excitation light concentrator and emission light collector,
>>>>one could design and use high NA immersion objective lenses with
>>>>better light throughput capabilities.
>>>>
>>>> 3. With incident/episcopic illumination, the amount of
>>>>image-contaminating background illumination light is greatly
>>>>reduced since most of the illumination light transmits through
>>>>the sample and never re-enters into the detection light path. As
>>>>stated in the Chroma Handbook of Optical Filters for Fluorescence
>>>>Microscopy, "... By illuminating with incident light [one needs
>>>>only to] filter out excitation light back-scattering from the
>>>>specimen or reflecting from glass surfaces . The use of
>>>>high-quality oil-immersion objectives (made with materials that
>>>>have minimal autofluorescence and using low-fluorescence oil)
>>>>eliminates surface reflections, which can reduce the level of
>>>>back-scattered light to as little as 1% of the incident light."
>>>>The quality of barrier filters employed at the time in early
>>>>diascopic fluorescence microscopes were such that they could not
>>>>achieve the same level of illumination light rejection in the
>>>>final image.
> >>>
>>>> 4. Early diascopic fluorescence microscopes used UV light to
>>>>excite fluorochromes/fluorescent probes embedded in a microscopic
>>>>sample. This design lessened the demands of the barrier filters
>>>>(UV light is absorbed by most glasses), but it also had the
>>>>disadvantage that the UV light could elicit autofluorescence in
>>>>the sample and optics of the microscope (which leads to image
>>>>background light again). In addition, the exciting UV light had
>>>>to traverse the sample and mounting slide, thus being absorbed
>>>>and scattered through thicker tissues and leading to weak
>>>>fluorescent image signals in those cases. As far as I can tell,
>>>>Ploem (and perhaps a few other researchers - still not clear to
>>>>me because I don't have access to the original research articles)
>>>>realized that common fluorescent dyes like FITC could be
>>>>efficiently excited with visible BLUE wavelengths and TRITC could
>>>>be efficiently excited with visible GREEN wavelengths, thereby
>>>>doing away with the need for pure UV excitation.
> >>>
>>>> 5. Bearing points 1-4 in mind, the implementation of the filter
>>>>cube block - the very heart of the epifluorescence microscope
>>>>design - now makes clear sense. The introduction of dichroic
>>>>beamsplitters by Brumberg, and their subsequent
>>>>commercialization/development by Ploem further improved the
>>>>filtering of excitation illumination light from the fluorescence
>>>>emission light and also created a convenient method of
>>>>introducing incident light onto the sample.
>>>>
>>>> It is often stated that the fluorescent signal that ultimately
>>>>forms the desired image of the sample is several orders of
>>>>magnitude weaker than the excitation light that is used to
>>>>generate it. That is to say, when we form an fluorescence image,
>>>>much effort has gone into filtering out and removing the
>>>>illumination light as much as possible to create a dark/black
>>>>background on which the fluorescence signal overlays. That's the
>>>>name of the game in fluorescence imaging - filter out the
>>>>unwanted signal as much as you can. It's not just the dichroic
>>>>mirror and barrier filters doing this. It's the incident light /
>>>>episcopic microscope design and the application of optimized
>>>>excitation wavelengths for the fluorescent probes that also play
>>>>a big role in this filtering process - a fact that I (and I
>>>>imagine most of us) take for granted every time we snap a
>>>>fluorescence image. It's a rather simple change to the microscope
>>>>that Ploem and his colleagues of the time made, but modern
>>>>fluorescence imaging (with confocal, TIRF, and super-resolution
>>>>methods) would not be what it is today with such widespread use
>>>>in research, medicine, and industry without that fundamental
>>>>change.
>>>>
>>>> And who could count the number of discoveries and advancements
>>>>that have come about with the fluorescence microscope since that
>>>>time? It is a true workhorse in biology and worthy of a Nobel in
>>>>my opinion then. I wish Dr. Ploem all the luck with the decision!
>>>>
>>>> John Oreopoulos
>>>> Staff Scientist
>>>> Spectral Applied Research
>>>> Richmond Hill, Ontario
>>>> Canada
>>>> www.spectral.ca
>>>>
>>>>
>>>> On 2013-04-21, at 11:13 PM, Guy Cox wrote:
>>>>
>>>>> *****
>>>>> To join, leave or search the confocal microscopy listserv, go to:
>>>>>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy>>>>> *****
>>>>>
>>>>> There is a historical essay on all this by Ploem and Walter,
>>>>>published by Leica in their series Scientific and Technical
>>>>>Information, Edition CDR 5, pp. 1-16,12/2001.
>>>>>"Multi-wavelength epi-illumination in fluorescence microscopy"
>>>>>
>>>>>
http://www.leica-microsystems.com/fileadmin/downloads/Other/Publications/Leica_STI_CDR5_ploem_walter_en.pdf.
>>>>>
>>>>> Brumberg is given due credit. Of course the Iron Curtain
>>>>>meant that Ploem was not originally aware of that work, and the
>>>>>Brumberg and Krylova 1953 paper is in Russian, so may not mean
>>>>>much to most of us even if it can be found. (Suspect you'd have
>>>>>to use Cyrillic Google to find since English Google doesn't).
>>>>>
>>>>> Guy
>>>>>
>>>>> -----Original Message-----
>>>>> From: Confocal Microscopy List
>>>>>[mailto:
[hidden email]] On Behalf Of Mark
>>>>>Cannell
> >>>> Sent: Monday, 22 April 2013 1:16 AM
>>>>> To:
[hidden email]
>>>>> Subject: Re: Inventor of fluorescence Ploemopak in running for
>>>>>Nobel Prize + website on his early technology
>>>>>
>>>>> *****
>>>>> To join, leave or search the confocal microscopy listserv, go to:
>>>>>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy>>>>> *****
>>>>>
>>>>> Hi John
>>>>>
>>>>> Here is a centenary review of his work....
>>>>>
http://link.springer.com/content/pdf/10.1134%2FS1062359007020161.pdf>>>>> Here is a list of papers that are available for a fee..
>>>>>
>>>>>
>>>>>
http://pubget.com/search?from=18912654&page=1&q=author%3A%22E+M+EM+BRUMBERG%22>>>>>
>>>>> perhaps you can get copies via your library?
>>>>>
>>>>> Cheers Mark
>>>>>
>>>>> On 20/04/2013, at 5:08 PM, John Oreopoulos
>>>>><
[hidden email]> wrote:
>>>>>
>>>>>> *****
>>>>>> To join, leave or search the confocal microscopy listserv, go to:
>>>>>>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> >>>>> *****
>>>>>>
>>>>>> Mark,
>>>>>>
>>>>>> Your last posting peaked my curiosity, so I decided to look a
>>>>>>bit into this. The best I could come up with was a document by
>>>>>>Barry Masters on the history of fluorescence microscopy:
>>>>>>
>>>>>>
>>>>>>
http://www.google.ca/url?sa=t&rct=j&q=&esrc=s&source=web&cd=11&ved=0CEMQFjAAOAo&url=http%3A%2F%2Fwww.fen.bilkent.edu.tr%2F~physics%2Fnews%2Fmasters%2FELS_Hist_Fl_Micro.pdf&ei=4rlyUe2hLtGp4APHr4GwCg&usg=AFQjCNEW1u-TzRGu5wml8GZ26qGUN9iW3A&sig2=HnzzQ9CvfTkEvfJWVcfCeg&bvm=bv.45512109,d.dmg&cad=rja>>>>>>
>>>>>> Both Brumberg and Ploem are mentioned in the context of some
>>>>>>very important developments of epi-fluorescence microscopy. By
>>>>>>chance, does anyone have a copy of the papers cited involving
>>>>>>these authors? (Brumberg 1959, and Ploem 1967).
>>>>>>
>>>>>> John Oreopoulos
>>>>>> Staff Scientist
>>>>>> Spectral Applied Research
>>>>>> Richmond Hill, Ontario
>>>>>> Canada
>>>>>> www.spectral.ca
>>>>>>
>>>>>>
>>>>>> On 2013-04-19, at 5:44 PM, Mark Cannell wrote:
>>>>>>
>>>>>>> *****
>>>>>>> To join, leave or search the confocal microscopy listserv, go to:
>>>>>>>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy>>>>>>> *****
>>>>>>>
>>>>>>> I hope the pioneering work in 1948 of Evengenii Mikhailovich
>>>>>>>Brumberg is mentioned/considered in this context.
>>>>>>>
>>>>>>> Cheers
>>>>>>>
>>>>>>>
[hidden email]
>>>>>
>>>>> Mark B. Cannell Ph.D. FRSNZ
>>>>> Professor of Cardiac Cell Biology
>>>>> School of Physiology & Pharmacology
>>>>> Medical Sciences Building
>>>>> University of Bristol
>>>>> Bristol
>>>>> BS8 1TD UK
>>>>>
>>>>>
[hidden email]
>>>
>>> Mark B. Cannell Ph.D. FRSNZ
>>> Professor of Cardiac Cell Biology
>>> School of Physiology & Pharmacology
>>> Medical Sciences Building
>>> University of Bristol
>>> Bristol
>>> BS8 1TD UK
>>>
>>>
[hidden email]
>>
>> Mark B. Cannell Ph.D. FRSNZ
>> Professor of Cardiac Cell Biology
>> School of Physiology & Pharmacology
>> Medical Sciences Building
>> University of Bristol
>> Bristol
>> BS8 1TD UK
>>
>>
[hidden email]
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