Inventor of fluorescence Ploemopak in running for Nobel Prize + website on his early technology

[Barbara Foster]

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Dear Fellow listers,

For those of you who do fluorescence, Dr Ploem, inventor of the early
epi-fluorescence system known as the Ploemopak, is in the running for
a Nobel Prize.  A special website has been set up on his
invention.  Below is a note he recently sent to the New York
Microscopical Society.

Enjoy!
Barbara Foster
Microscopy/Microscopy Education
[hidden email]
[phone]
User Study now running: New Directions in Live Cell & Tissue Imaging
April 17-26.  Visit www.MicroscopyEducation.com for details


Dear Mel Pollinger,
I have been mentioned on the Dutch media as a possible
candidate for a Nobel Price. Some members of the N.Y.M.S may be
interested in some details on the development of multi-wavelenghs
epifluorescence microscopy.

A comprehensive WEBSITE has been created for the National Dutch
Science Museum to illustrate the reason for the incorporation of my
early prototype fluorescence epiilluminators in the permanent
collection of that museum, as well as the exposition of the very
first Leitz-Leica epi-illumination fluorescence microscope with a
Ploemopak fluorescence epi-illuminator in the VISEUM science Museum
in Wetzlar. Germany.

Website:
www.ploem-fluorescence-microscopy.com
With kind regards
Johan Ploem, (J.S. Ploem)

[Mark Cannell-2]

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I hope the pioneering work in 1948 of Evengenii Mikhailovich Brumberg is mentioned/considered in this context.

Cheers

[hidden email]

[John Oreopoulos]

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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]

[George McNamara]

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On 4/20/2013 11:08 AM, John Oreopoulos wrote:

F.W.D. Rost published three books with extensive (occasionally tedious)
history of fluorescence microscopy.

Page 187 to 188 of Fluorescence Microscopy volume 2:

http://books.google.com/books?id=-eJsstG_ClcC&printsec=frontcover&dq=Fluorescence+microscopy.+2&hl=en&sa=X&ei=ZcVyUerSOoy89QTak4GABQ&ved=0CDwQ6AEwAA#v=onepage&q=Fluorescence%20microscopy.%202&f=false 




1992 Fluorescence Microscopy - volume 1
http://books.google.com/books/about/Fluorescence_microscopy_1.html?id=RU_6doL1q0IC 


    Fluorescence microscopy is used for studying the distribution of
    substances which are present in very small amounts. The high
    sensitivity of the method makes it ideal for studying the
    distribution of substances in living cells. Its techniques are used
    mainly in biology and medicine, but are also valuable in coal
    petrology and elsewhere. The best-known application is in
    immunofluorescence. This magnificent new work provides comprehensive
    coverage of all aspects of fluorescence microscopy. It covers
    instrumentation, applications to a wide variety of fields, and the
    history of the technique. There is a chapter on quantitative
    techniques, including scanning: this aspect is dealt with in more
    detail in a companion volume, Quantitative Fluorescence Microscopy.
    Volume 1 deals with instrumentation and techniques for fluorescence
    microscopy, and includes the chapter on quantitation and scanning.
    Volume 2 deals with the applications of fluorescence microscopy in
    many fields. It includes information on fluorochromes and on
    autofluorescence. An invaluable appendix provides an alphabetical
    list of fluorochromes, giving information concerning chemical
    structure, fluorescence properties, applications and suitable filter
    combinations. These two important volumes will be of use to all
    fluorescence microscopists and will be an invaluable reference tool
    for those graduate students and research workers in biology,
    medicine and earth science who need to make use of these techniques.


1991 Quantitative fluorescence microscopy
http://books.google.com/books?id=G3cC7WQMArYC&source=gbs_similarbooks

    Quantitative fluorescence microscopy is concerned with making
    measurements from fluorescent specimens in a fluorescence
    microscope, by measuring fluorescence emission from a defined area
    or areas of a specimen. This technique is most commonly used to
    determine the amount of some specific substance, such as DNA, in
    some particular area of a cell. But it has many other uses; for
    example, it can be used to identify certain substances in the cell
    by examining their fluorescence characteristics. This book is a
    complete guide to this technique for all biologists. It describes
    the principles and applications of quantitative fluorescence
    microscopy and also gives much practical information about the
    instrumentation required. There is also a discussion of the exciting
    developments in confocal fluorescence microscopy which allows the
    three dimensional distribution of particular substances to be
    determined. Everyone presently using this technique, or wishing to
    start using it will need to read this book.


Enjoy,

George

> *****
> 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]
>>      
>    

[Guy Cox-2]

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Oh dear, nostalgia.  Fred Rost was a wonderful microscopist and a great professor (Anatomy, University of NSW) but a bit off-putting at first encounter since he always dressed (totally unconvincingly) as a woman.  But yes, he certainly knew the history of fluorescence microscopy.

                                                                       Guy

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of George McNamara
Sent: Sunday, 21 April 2013 2:58 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
*****

On 4/20/2013 11:08 AM, John Oreopoulos wrote:

F.W.D. Rost published three books with extensive (occasionally tedious) history of fluorescence microscopy.

Page 187 to 188 of Fluorescence Microscopy volume 2:

http://books.google.com/books?id=-eJsstG_ClcC&printsec=frontcover&dq=Fluorescence+microscopy.+2&hl=en&sa=X&ei=ZcVyUerSOoy89QTak4GABQ&ved=0CDwQ6AEwAA#v=onepage&q=Fluorescence%20microscopy.%202&f=false 




1992 Fluorescence Microscopy - volume 1
http://books.google.com/books/about/Fluorescence_microscopy_1.html?id=RU_6doL1q0IC 


    Fluorescence microscopy is used for studying the distribution of
    substances which are present in very small amounts. The high
    sensitivity of the method makes it ideal for studying the
    distribution of substances in living cells. Its techniques are used
    mainly in biology and medicine, but are also valuable in coal
    petrology and elsewhere. The best-known application is in
    immunofluorescence. This magnificent new work provides comprehensive
    coverage of all aspects of fluorescence microscopy. It covers
    instrumentation, applications to a wide variety of fields, and the
    history of the technique. There is a chapter on quantitative
    techniques, including scanning: this aspect is dealt with in more
    detail in a companion volume, Quantitative Fluorescence Microscopy.
    Volume 1 deals with instrumentation and techniques for fluorescence
    microscopy, and includes the chapter on quantitation and scanning.
    Volume 2 deals with the applications of fluorescence microscopy in
    many fields. It includes information on fluorochromes and on
    autofluorescence. An invaluable appendix provides an alphabetical
    list of fluorochromes, giving information concerning chemical
    structure, fluorescence properties, applications and suitable filter
    combinations. These two important volumes will be of use to all
    fluorescence microscopists and will be an invaluable reference tool
    for those graduate students and research workers in biology,
    medicine and earth science who need to make use of these techniques.


1991 Quantitative fluorescence microscopy http://books.google.com/books?id=G3cC7WQMArYC&source=gbs_similarbooks

    Quantitative fluorescence microscopy is concerned with making
    measurements from fluorescent specimens in a fluorescence
    microscope, by measuring fluorescence emission from a defined area
    or areas of a specimen. This technique is most commonly used to
    determine the amount of some specific substance, such as DNA, in
    some particular area of a cell. But it has many other uses; for
    example, it can be used to identify certain substances in the cell
    by examining their fluorescence characteristics. This book is a
    complete guide to this technique for all biologists. It describes
    the principles and applications of quantitative fluorescence
    microscopy and also gives much practical information about the
    instrumentation required. There is also a discussion of the exciting
    developments in confocal fluorescence microscopy which allows the
    three dimensional distribution of particular substances to be
    determined. Everyone presently using this technique, or wishing to
    start using it will need to read this book.


Enjoy,

George

> *****
> 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=0CE
> MQFjAAOAo&url=http%3A%2F%2Fwww.fen.bilkent.edu.tr%2F~physics%2Fnews%2F
> masters%2FELS_Hist_Fl_Micro.pdf&ei=4rlyUe2hLtGp4APHr4GwCg&usg=AFQjCNEW
> 1u-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 Cannell-2]

*****
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]

[Guy Cox-2]

*****
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]

[John Oreopoulos]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Another member of this listserver has kindly provided me with a scanned copy of the Brumberg paper from 1959 (Biophysics), and it translated into English. It is a very interesting historical reading, and if anyone else is interested, I can send a copy to you offline.

Cheers,

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]

[mmodel]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

I wrote to a colleague in Russia, he still keeps an objective built by Brumberg. I wonder if any museum could be interested in this?

________________________________________
From: Confocal Microscopy List [[hidden email]] on behalf of John Oreopoulos [[hidden email]]
Sent: Monday, April 22, 2013 4:06 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
*****

Another member of this listserver has kindly provided me with a scanned copy of the Brumberg paper from 1959 (Biophysics), and it translated into English. It is a very interesting historical reading, and if anyone else is interested, I can send a copy to you offline.

Cheers,

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]

[Dmitry Sokolov]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Thank you John,
great job!

Dear Guy, thank you for the link. Leica, however, has given a wrong
citation to the Brumberg's paper. The correct citations can be found here:
http://confocal-manawatu.pbworks.com/w/page/65550417/First%20Paper%20on%20Fluorescence%20Microscopy

Regarding the Global priority of any particular knowledge and IP, I
believe that the "barriers" similar to the Iron Curtain were still
transparent to the scientific papers published in the open journals.
 From the other hand, "lack of the access to the Intellectual Property
produced in a local language cannot be the excuse for postulating the
Global priority in this particular IP produced later in another local
language. Principles of Equity of language and geographic location can
be observed if based on the chronological priority only."
http://confocal-manawatu.pbworks.com/w/page/65615734/Local%20vs%20Global%20Priority%20of%20Intellectual%20Property

As the examples, the atomic bombing of Japan by USA and the first manned
space flight by Soviets are the well-known evidence of the historical
priority in the top secret IPs respectively at that time.

I believe that the translation to English and sharing of the
historically interesting artefacts like first paper on epifluorescence
microscopy is a minor problem and can be easily crowdsourced in MIAWiki
or elsewhere:
http://confocal-manawatu.pbworks.com/w/page/60427499/What%20is%20MIAWiki%20About

Cheers,
Dmitry

*Advanced Knowledge Management*
for *MICROSCOPY *and *Image Analysis *
------------------------------------------------------------------------
*Dmitry Sokolov*, Ph.D.
Mob: *[phone]***
[hidden email] <mailto:[hidden email]>

On 23.04.2013 8:06, John Oreopoulos wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Another member of this listserver has kindly provided me with a scanned copy of the Brumberg paper from 1959 (Biophysics), and it translated into English. It is a very interesting historical reading, and if anyone else is interested, I can send a copy to you offline.
>
> Cheers,
>
> 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]

[John Oreopoulos]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Thanks should instead be given to Jolien Tyler who took the time to journey to some deep, dark corner of her university library and find this paper on my request.

John Oreopoulos


On 2013-04-23, at 8:57 AM, Dmitry Sokolov wrote:

> Thank you John,
> great job!
>
> Dear Guy, thank you for the link. Leica, however, has given a wrong citation to the Brumberg's paper. The correct citations can be found here:
> http://confocal-manawatu.pbworks.com/w/page/65550417/First%20Paper%20on%20Fluorescence%20Microscopy
>
> Regarding the Global priority of any particular knowledge and IP, I believe that the "barriers" similar to the Iron Curtain were still transparent to the scientific papers published in the open journals. From the other hand, "lack of the access to the Intellectual Property produced in a local language cannot be the excuse for postulating the Global priority in this particular IP produced later in another local language. Principles of Equity of language and geographic location can be observed if based on the chronological priority only."
> http://confocal-manawatu.pbworks.com/w/page/65615734/Local%20vs%20Global%20Priority%20of%20Intellectual%20Property
>
> As the examples, the atomic bombing of Japan by USA and the first manned space flight by Soviets are the well-known evidence of the historical priority in the top secret IPs respectively at that time.
>
> I believe that the translation to English and sharing of the historically interesting artefacts like first paper on epifluorescence microscopy is a minor problem and can be easily crowdsourced in MIAWiki or elsewhere:
> http://confocal-manawatu.pbworks.com/w/page/60427499/What%20is%20MIAWiki%20About
>
> Cheers,
> Dmitry
>
> Advanced Knowledge Management
> for MICROSCOPY and Image Analysis
> Dmitry Sokolov, Ph.D.
> Mob: [phone]
> [hidden email]
>
> On 23.04.2013 8:06, John Oreopoulos wrote:
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Another member of this listserver has kindly provided me with a scanned copy of the Brumberg paper from 1959 (Biophysics), and it translated into English. It is a very interesting historical reading, and if anyone else is interested, I can send a copy to you offline.
>>
>> Cheers,
>>
>> 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]
>

[John Oreopoulos]

*****
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 Cannell-2]

*****
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]

[Guy Cox-2]

*****
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 Cannell-2]

*****
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]

[John Oreopoulos]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Mark, if I'm not mistaken, have there not been Nobels awarded for phase contrast microscopy and other inventions like the CCD camera? So there are examples of tool builders for science being awarded this prize.

But it is unfortunate that the prize can only be split a maximum between three people.

John Oreopoulos

On 2013-04-25, 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]

[Tim Feinstein-2]

*****
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]

[Guy Cox-2]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Well, whatever, but what a technique!  Few things have changed the world as much as that.

                                                         Guy

     

-----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]

[Martin Wessendorf-2]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

On 4/25/2013 4:19 AM, Guy Cox wrote:

> I do think that the pioneers of immuno-fluorescence also deserve a Nobel!

Now THERE is a prize that would've been worth presenting!  Sadly, Albert
Coons, the father of immunofluorescence, died in 1978, but there are
other worthies who contributed to the widespread usefulness of the
technique.

Having used a fluorescence microscope with a sub-stage darkfield
condenser, I can say that the system has at least one advantage: the
brightness of the image was (partly) a function of the NA of that
condenser.  If you had dim labeling and want to get a low-mag image of
it, diascopic illumination was a good way to go.

Martin Wessendorf


--
Martin Wessendorf, Ph.D.                   office: [phone]
Assoc Prof, Dept Neuroscience                 lab: [phone]
University of Minnesota             Preferred FAX: [phone]
6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
Minneapolis, MN  55455                    e-mail: [hidden email]

[James Pawley]

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To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Well, whatever, but what a technique!  Few things have changed the
>world as much as that.
>
>                                                          Guy
>
>

Just to stir a little, Don Glaser got the 1960 Physics prize for the
hydrogen bubble chamber and Ruska finally got the 1988 prize for the
electron microscope, something that he first demonstrated in 1933. He
had to share it with two upstarts who had just come up with the idea
for scanning tunnelling microscope.

Looking around, I would guess that epi-fluorescence is the source of
more useful information than any of these.

Cheers,

Jim Pawley

>    
>
>-----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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