Mounting media for STED

[Craig Brideau]

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*****

I started working with a WLL many years ago with a Fianium (I think NKT
bought them out) continuum optimized for early STED. The power spectral
density of WLL is fairly variable across the entire spectral range; the
very early model I was using went from 0.1mW/nm at 440 gradually increasing
to 1.0mW/nm at 600 to ~900nm. Back then, to even get 0.1mW/nm at 440 the
company had to cherry-pick a non-linear fiber for me to get one that was
that short. Their typical model usually only produced power starting at
470nm. I'm pleased to see the technology advancing to shift more power into
blue end of the spectrum compared to the earlier models, and would love to
see the mW/nm power spectral density curves of the various options. These
give you some idea, at a glance, of what performance you can expect within
a given wavelength band.

Craig

On Tue, Apr 14, 2020 at 10:42 AM Vitaly Boyko <
[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
>  I asked for 440-750 nm, and they did 440-790 for the WLL. At least Leica
> DOES listen!!! ... compared to others, including "fake" multiplex by the
> "neighbor"... Stellaris will be very useful for what we are currently doing
> at our facility!!! Cheers,VitalyP.S. Sarcastic joke: If even Vitaly likes
> it, it is a big step forward!!!
>
>     On Tuesday, April 14, 2020, 11:03:54 AM EDT, Christoph Ruediger Bauer <
> [hidden email]> wrote:
>
>  *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear Ralph,
> I think you got this one wrong. By curiosity I googled Arvydas facility
> and found this: "Core: Advanced Fluorescent Imaging Core Core Director:
> Arvydas Matiukas, PhD". From looking at his site (
> https://www.upstate.edu/research/facilities/confocal.php) it seems clear
> to me that this is not a Leica facility and his list of core facility
> instruments starts actually with two Zeiss confocals.
> I think your conclusions concerning a commercial mailing were a bit fast.
> Best wishes,
> Christoph
>
>
> BIOIMAGING CENTER
> University of Geneva - Science II
> Room 245
> 30, Quai Ernest Ansermet
> CH - 1211 Genève 4
>
> Dr. Christoph R. Bauer
> Managing Director
> Tel.: + 41 22 3796632
> Fax: + 41 22 3796868
> email: [hidden email]
> website: http://bioimaging.unige.ch/
>
>
>
> -----Original Message-----
> From: Confocal Microscopy List <[hidden email]> On
> Behalf Of Ralf Palmisano
> Sent: 14 April 2020 15:41
> To: [hidden email]
> Subject: Re: New Leica Stellaris vs SP8
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi Director Arvydas of LEICA Microscopy Core,
>
> This mail rang so many bells while reading. It is this sort of masked
> commercial mailings that I don't like to see here.
> I was wondering why this initial mail was subjected a re-post. In fact it
> was not are-post. It is a sort of undercover commercial post.
> Why are you hiding that this is the initial post?
> Why are you hiding that you are director of Leica microscopy core.
> Why do make me search online who "Director of Microscopy Core" is?
>
> I don't think that Leica supports this type of misguiding this community,
> by pretending to be sort of a fair balanced user and starting a promotional
> tour.
>
> Don't get me wrong. This is not about Leica pro or con. This is about your
> rather shabby behaviour here.
> Don't get me wrong. If you would have made yourself known to be a Leica
> sponsored facility manager and asked for the communities opion, well fair
> enough, then everys would be aware what one is stepping into.
>
> Leica Microsystems deserve much better than people like you coming in such
> questionable disguise (yes, that's a Shakespeare quote). Apart from it,
> Leica would certainly have phrased this commercial way better than you,
> Sir, ever will be able, too.
>
> I sincerely apologise if you think this was personal, it is not and I
> believe this sort of posts always prove to be of disservice for the
> respective company.
>
> Have a nice day,
>
> Ralph Palmisano
> Head Optical Imaging Centre Erlangen
> Central Institute
> Friedrich-Alexander University of Erlangen-Nuremberg
>
> Am 14/04/2020 um 14:40 schrieb Arvydas Matiukas:
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > Hello Microscopists,
> >
> >
> > What do you think of the last week presented Leica Stellaris new
> generation of confocal/STED microscopes? I was planning to buy SP8 but now
> I think it is worth to wait till Stellaris is available. Second generation
> WLL with 350 nm range and most sensitiive new HyD detectors are worth it.
> >
> >
> > Thanks,
> > Arvydas
> >
> >
> > Director of Microscopy Core
>
> --
> Ralph Palmisano
> Head - Optical Imaging Centre Erlangen
>
> Fellow Royal Microscopical Society
> Member Royal Society of Medicine
>
> Speaker Scientific Advisory Board "German BioImaging"
> Board of Directors Core Technologies for Life Science
>
> Cauerstr. 3
> 91058 Erlangen, Germany
>
> [phone] (Office)
> [phone] (Secretary)
>
> www.oice.uni-erlangen.de
>
>

[Mariette Dupont]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear Vitaly,

That's great to hear! As Leica seems to listen to you, could you please ask
them to explain the quantitativeness of their Thunder processing - well,
maybe start with "How does it actually really work?", as well as the
strange sudden comparison of a detector's PDE, where everyone else refers
to QE (what's to hide?) or the referral to 420fps without mentioning any
frame-size, resolution and so on? And how about, as Craig mentioned, the
power-distribution of the WLL at sample level?

I agree that the system does look pretty nice, honestly, so does the WLL
and to some extent, the FLIM implementation - even though as most of us
realize, FLIM does require in-depth knowledge of its data analysis and is
not nor will never be as straight-forward as presented. Yet, easier
implementation may make it available to new users. I, however, fear FLIM
data to be horribly misinterpreted and I for one do not want to be referred
to papers being withdrawn because of this inexperience.
.
But all that aside, I'm disappointed to see your reaction and how you drag
in other companies without any support of your statements nor links to the
current discussion. I have not been active yet on this channel, more of a
passive reader/follower while learning a lot, but would be much more
delighted to see this channel used to discuss and propose improvements,
rather than unknowingly bashing on a company because of not fully
comprehending the technology, disagreement to their approach to supporting
science, or simply because you aim to be a spokesman for a competitor
company. I'm convinced all companies to put a lot of time & money in
new developments, and constructive criticism would be much more useful than
negative-but-void opinions. A researcher, and with that, facility manager,
is, in my opinion, supposed to be objective, at all times.

Marie

On Tue, Apr 14, 2020 at 8:03 PM Craig Brideau <[hidden email]>
wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> I started working with a WLL many years ago with a Fianium (I think NKT
> bought them out) continuum optimized for early STED. The power spectral
> density of WLL is fairly variable across the entire spectral range; the
> very early model I was using went from 0.1mW/nm at 440 gradually increasing
> to 1.0mW/nm at 600 to ~900nm. Back then, to even get 0.1mW/nm at 440 the
> company had to cherry-pick a non-linear fiber for me to get one that was
> that short. Their typical model usually only produced power starting at
> 470nm. I'm pleased to see the technology advancing to shift more power into
> blue end of the spectrum compared to the earlier models, and would love to
> see the mW/nm power spectral density curves of the various options. These
> give you some idea, at a glance, of what performance you can expect within
> a given wavelength band.
>
> Craig
>
> On Tue, Apr 14, 2020 at 10:42 AM Vitaly Boyko <
> [hidden email]> wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> >  I asked for 440-750 nm, and they did 440-790 for the WLL. At least Leica
> > DOES listen!!! ... compared to others, including "fake" multiplex by the
> > "neighbor"... Stellaris will be very useful for what we are currently
> doing
> > at our facility!!! Cheers,VitalyP.S. Sarcastic joke: If even Vitaly likes
> > it, it is a big step forward!!!
> >
> >     On Tuesday, April 14, 2020, 11:03:54 AM EDT, Christoph Ruediger
> Bauer <
> > [hidden email]> wrote:
> >
> >  *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > Dear Ralph,
> > I think you got this one wrong. By curiosity I googled Arvydas facility
> > and found this: "Core: Advanced Fluorescent Imaging Core Core Director:
> > Arvydas Matiukas, PhD". From looking at his site (
> > https://www.upstate.edu/research/facilities/confocal.php) it seems clear
> > to me that this is not a Leica facility and his list of core facility
> > instruments starts actually with two Zeiss confocals.
> > I think your conclusions concerning a commercial mailing were a bit fast.
> > Best wishes,
> > Christoph
> >
> >
> > BIOIMAGING CENTER
> > University of Geneva - Science II
> > Room 245
> > 30, Quai Ernest Ansermet
> > CH - 1211 Genève 4
> >
> > Dr. Christoph R. Bauer
> > Managing Director
> > Tel.: + 41 22 3796632
> > Fax: + 41 22 3796868
> > email: [hidden email]
> > website: http://bioimaging.unige.ch/
> >
> >
> >
> > -----Original Message-----
> > From: Confocal Microscopy List <[hidden email]> On
> > Behalf Of Ralf Palmisano
> > Sent: 14 April 2020 15:41
> > To: [hidden email]
> > Subject: Re: New Leica Stellaris vs SP8
> >
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > Hi Director Arvydas of LEICA Microscopy Core,
> >
> > This mail rang so many bells while reading. It is this sort of masked
> > commercial mailings that I don't like to see here.
> > I was wondering why this initial mail was subjected a re-post. In fact it
> > was not are-post. It is a sort of undercover commercial post.
> > Why are you hiding that this is the initial post?
> > Why are you hiding that you are director of Leica microscopy core.
> > Why do make me search online who "Director of Microscopy Core" is?
> >
> > I don't think that Leica supports this type of misguiding this community,
> > by pretending to be sort of a fair balanced user and starting a
> promotional
> > tour.
> >
> > Don't get me wrong. This is not about Leica pro or con. This is about
> your
> > rather shabby behaviour here.
> > Don't get me wrong. If you would have made yourself known to be a Leica
> > sponsored facility manager and asked for the communities opion, well fair
> > enough, then everys would be aware what one is stepping into.
> >
> > Leica Microsystems deserve much better than people like you coming in
> such
> > questionable disguise (yes, that's a Shakespeare quote). Apart from it,
> > Leica would certainly have phrased this commercial way better than you,
> > Sir, ever will be able, too.
> >
> > I sincerely apologise if you think this was personal, it is not and I
> > believe this sort of posts always prove to be of disservice for the
> > respective company.
> >
> > Have a nice day,
> >
> > Ralph Palmisano
> > Head Optical Imaging Centre Erlangen
> > Central Institute
> > Friedrich-Alexander University of Erlangen-Nuremberg
> >
> > Am 14/04/2020 um 14:40 schrieb Arvydas Matiukas:
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > > Post images on http://www.imgur.com and include the link in your
> > posting.
> > > *****
> > >
> > > Hello Microscopists,
> > >
> > >
> > > What do you think of the last week presented Leica Stellaris new
> > generation of confocal/STED microscopes? I was planning to buy SP8 but
> now
> > I think it is worth to wait till Stellaris is available. Second
> generation
> > WLL with 350 nm range and most sensitiive new HyD detectors are worth it.
> > >
> > >
> > > Thanks,
> > > Arvydas
> > >
> > >
> > > Director of Microscopy Core
> >
> > --
> > Ralph Palmisano
> > Head - Optical Imaging Centre Erlangen
> >
> > Fellow Royal Microscopical Society
> > Member Royal Society of Medicine
> >
> > Speaker Scientific Advisory Board "German BioImaging"
> > Board of Directors Core Technologies for Life Science
> >
> > Cauerstr. 3
> > 91058 Erlangen, Germany
> >
> > [phone] (Office)
> > [phone] (Secretary)
> >
> > www.oice.uni-erlangen.de
> >
> >
>

[Vitaly Boyko-2]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

 Hi Marie,
we do not rely on Thunder, we rely on experimentally obtained Point-Spread Function and a 3rd party deconvolution software. It is extremely important to learn on how to obtain the best possible and the most accurate PSF experimentally.  There are no issues with Leica FLIM, it works well, no complains, we got very nice data the the flipper, a membrane sensor. Also, we were able to separate Alexa 488 and CF488, or Alexa 568 and Alexa 594. People have to learn and understand phasor. It is relatively easy to learn, my 11.75 years old son knows the basics of phasor already.
Good luck,
Vitaly 
  
    On Tuesday, April 14, 2020, 04:03:38 PM EDT, Mariette Dupont <[hidden email]> wrote:  
 
 *****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear Vitaly,

That's great to hear! As Leica seems to listen to you, could you please ask
them to explain the quantitativeness of their Thunder processing - well,
maybe start with "How does it actually really work?", as well as the
strange sudden comparison of a detector's PDE, where everyone else refers
to QE (what's to hide?) or the referral to 420fps without mentioning any
frame-size, resolution and so on? And how about, as Craig mentioned, the
power-distribution of the WLL at sample level?

I agree that the system does look pretty nice, honestly, so does the WLL
and to some extent, the FLIM implementation - even though as most of us
realize, FLIM does require in-depth knowledge of its data analysis and is
not nor will never be as straight-forward as presented. Yet, easier
implementation may make it available to new users. I, however, fear FLIM
data to be horribly misinterpreted and I for one do not want to be referred
to papers being withdrawn because of this inexperience.
.
But all that aside, I'm disappointed to see your reaction and how you drag
in other companies without any support of your statements nor links to the
current discussion. I have not been active yet on this channel, more of a
passive reader/follower while learning a lot, but would be much more
delighted to see this channel used to discuss and propose improvements,
rather than unknowingly bashing on a company because of not fully
comprehending the technology, disagreement to their approach to supporting
science, or simply because you aim to be a spokesman for a competitor
company. I'm convinced all companies to put a lot of time & money in
new developments, and constructive criticism would be much more useful than
negative-but-void opinions. A researcher, and with that, facility manager,
is, in my opinion, supposed to be objective, at all times.

Marie

On Tue, Apr 14, 2020 at 8:03 PM Craig Brideau <[hidden email]>
wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> I started working with a WLL many years ago with a Fianium (I think NKT
> bought them out) continuum optimized for early STED. The power spectral
> density of WLL is fairly variable across the entire spectral range; the
> very early model I was using went from 0.1mW/nm at 440 gradually increasing
> to 1.0mW/nm at 600 to ~900nm. Back then, to even get 0.1mW/nm at 440 the
> company had to cherry-pick a non-linear fiber for me to get one that was
> that short. Their typical model usually only produced power starting at
> 470nm. I'm pleased to see the technology advancing to shift more power into
> blue end of the spectrum compared to the earlier models, and would love to
> see the mW/nm power spectral density curves of the various options. These
> give you some idea, at a glance, of what performance you can expect within
> a given wavelength band.
>
> Craig
>
> On Tue, Apr 14, 2020 at 10:42 AM Vitaly Boyko <
> [hidden email]> wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> >  I asked for 440-750 nm, and they did 440-790 for the WLL. At least Leica
> > DOES listen!!! ... compared to others, including "fake" multiplex by the
> > "neighbor"... Stellaris will be very useful for what we are currently
> doing
> > at our facility!!! Cheers,VitalyP.S. Sarcastic joke: If even Vitaly likes
> > it, it is a big step forward!!!
> >
> >    On Tuesday, April 14, 2020, 11:03:54 AM EDT, Christoph Ruediger
> Bauer <
> > [hidden email]> wrote:
> >
> >  *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > Dear Ralph,
> > I think you got this one wrong. By curiosity I googled Arvydas facility
> > and found this: "Core: Advanced Fluorescent Imaging Core Core Director:
> > Arvydas Matiukas, PhD". From looking at his site (
> > https://www.upstate.edu/research/facilities/confocal.php) it seems clear
> > to me that this is not a Leica facility and his list of core facility
> > instruments starts actually with two Zeiss confocals.
> > I think your conclusions concerning a commercial mailing were a bit fast.
> > Best wishes,
> > Christoph
> >
> >
> > BIOIMAGING CENTER
> > University of Geneva - Science II
> > Room 245
> > 30, Quai Ernest Ansermet
> > CH - 1211 Genève 4
> >
> > Dr. Christoph R. Bauer
> > Managing Director
> > Tel.: + 41 22 3796632
> > Fax: + 41 22 3796868
> > email: [hidden email]
> > website: http://bioimaging.unige.ch/
> >
> >
> >
> > -----Original Message-----
> > From: Confocal Microscopy List <[hidden email]> On
> > Behalf Of Ralf Palmisano
> > Sent: 14 April 2020 15:41
> > To: [hidden email]
> > Subject: Re: New Leica Stellaris vs SP8
> >
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > Hi Director Arvydas of LEICA Microscopy Core,
> >
> > This mail rang so many bells while reading. It is this sort of masked
> > commercial mailings that I don't like to see here.
> > I was wondering why this initial mail was subjected a re-post. In fact it
> > was not are-post. It is a sort of undercover commercial post.
> > Why are you hiding that this is the initial post?
> > Why are you hiding that you are director of Leica microscopy core.
> > Why do make me search online who "Director of Microscopy Core" is?
> >
> > I don't think that Leica supports this type of misguiding this community,
> > by pretending to be sort of a fair balanced user and starting a
> promotional
> > tour.
> >
> > Don't get me wrong. This is not about Leica pro or con. This is about
> your
> > rather shabby behaviour here.
> > Don't get me wrong. If you would have made yourself known to be a Leica
> > sponsored facility manager and asked for the communities opion, well fair
> > enough, then everys would be aware what one is stepping into.
> >
> > Leica Microsystems deserve much better than people like you coming in
> such
> > questionable disguise (yes, that's a Shakespeare quote). Apart from it,
> > Leica would certainly have phrased this commercial way better than you,
> > Sir, ever will be able, too.
> >
> > I sincerely apologise if you think this was personal, it is not and I
> > believe this sort of posts always prove to be of disservice for the
> > respective company.
> >
> > Have a nice day,
> >
> > Ralph Palmisano
> > Head Optical Imaging Centre Erlangen
> > Central Institute
> > Friedrich-Alexander University of Erlangen-Nuremberg
> >
> > Am 14/04/2020 um 14:40 schrieb Arvydas Matiukas:
> > > *****
> > > To join, leave or search the confocal microscopy listserv, go to:
> > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > > Post images on http://www.imgur.com and include the link in your
> > posting.
> > > *****
> > >
> > > Hello Microscopists,
> > >
> > >
> > > What do you think of the last week presented Leica Stellaris new
> > generation of confocal/STED microscopes? I was planning to buy SP8 but
> now
> > I think it is worth to wait till Stellaris is available. Second
> generation
> > WLL with 350 nm range and most sensitiive new HyD detectors are worth it.
> > >
> > >
> > > Thanks,
> > > Arvydas
> > >
> > >
> > > Director of Microscopy Core
> >
> > --
> > Ralph Palmisano
> > Head - Optical Imaging Centre Erlangen
> >
> > Fellow Royal Microscopical Society
> > Member Royal Society of Medicine
> >
> > Speaker Scientific Advisory Board "German BioImaging"
> > Board of Directors Core Technologies for Life Science
> >
> > Cauerstr. 3
> > 91058 Erlangen, Germany
> >
> > [phone] (Office)
> > [phone] (Secretary)
> >
> > www.oice.uni-erlangen.de
> >
> >
>