Phototoxicity

[Erik Manders-3]

*****
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 all,

As you may know It has been a while ago that I have worked on methods to reduce the toxic effects of excitation light in fluorescence microscopy. When we worked on Controlled Light Exposure Microscopy (nowadays called Spatially Controlled Illumination Microscopy in order to reduce confusion...) there was not much know about the photo-physical background of phototoxicity. There wasn't even any paper describing dose-effect of light in microscopy.

Simple questions like:
- How much light kills a cell?
- How much light stops the cell cycle without killing the cell?
- What is the role of the fluorophore?
- Are some fluorophores more phototoxic than others?
- Are oxygen radicals directly involved or singlet oxygen?
- Has dose-rate (at constant dose) an influence on cell happiness?
- A thousand other questions, but the main simple question is: How much light kills a cell???

Who has an answer to these questions or knows the title of a paper that goed into this kind of detail. I am out of this subject for some years now, but I am very interested to get an update and see if any progress has been made since the end 90s.

Kind regards, Erik Manders

[phil laissue-3]

*****
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 Erik,
for an update, there's Icha et al. 2017 (PMID: 28749075). We cited a lot of
relevant papers in ours (PMID: 28661494 - particularly in the supplementary
notes) and show a reduction in GMK proliferation, without killing cells, at
low dose & irradiance. Cranfill et al. 2016 have demonstrated accelerated
photobleaching (PMID: 27240257), and Waeldchen et al. 2015 (PMID: 26481189)
show that it really depends on what type of cell (e.g. HeLa can take a
beating).
There's many more, but this is a good start. Please let me know if you
don't have access to any of the papers, I can send them directly to you.
With kind regards,
Philippe
_____________________________________
Philippe Laissue, PhD
Lecturer in Bioimaging
School of Biological Sciences, Room 4.17
University of Essex, Colchester CO4 3SQ, UK
(0044) 01206 872246 / [phone] 6056
[hidden email]
website <https://laissue.github.io/>


On Wed, 10 Jun 2020 at 15:15, Erik Manders <[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 all,
>
> As you may know It has been a while ago that I have worked on methods to
> reduce the toxic effects of excitation light in fluorescence microscopy.
> When we worked on Controlled Light Exposure Microscopy (nowadays called
> Spatially Controlled Illumination Microscopy in order to reduce
> confusion...) there was not much know about the photo-physical background
> of phototoxicity. There wasn't even any paper describing dose-effect of
> light in microscopy.
>
> Simple questions like:
> - How much light kills a cell?
> - How much light stops the cell cycle without killing the cell?
> - What is the role of the fluorophore?
> - Are some fluorophores more phototoxic than others?
> - Are oxygen radicals directly involved or singlet oxygen?
> - Has dose-rate (at constant dose) an influence on cell happiness?
> - A thousand other questions, but the main simple question is: How much
> light kills a cell???
>
> Who has an answer to these questions or knows the title of a paper that
> goed into this kind of detail. I am out of this subject for some years now,
> but I am very interested to get an update and see if any progress has been
> made since the end 90s.
>
> Kind regards, Erik Manders
>

[lechristophe]

*****
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 Erik,

I'd add a recent review from Ricardo Henriques' lab that contains a lot of
useful references:
https://iopscience.iop.org/article/10.1088/1361-6463/ab6b95

Christophe



On Wed, 10 Jun 2020 at 17:26, phil laissue <
[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi Erik,
> for an update, there's Icha et al. 2017 (PMID: 28749075). We cited a lot of
> relevant papers in ours (PMID: 28661494 - particularly in the supplementary
> notes) and show a reduction in GMK proliferation, without killing cells, at
> low dose & irradiance. Cranfill et al. 2016 have demonstrated accelerated
> photobleaching (PMID: 27240257), and Waeldchen et al. 2015 (PMID: 26481189)
> show that it really depends on what type of cell (e.g. HeLa can take a
> beating).
> There's many more, but this is a good start. Please let me know if you
> don't have access to any of the papers, I can send them directly to you.
> With kind regards,
> Philippe
> _____________________________________
> Philippe Laissue, PhD
> Lecturer in Bioimaging
> School of Biological Sciences, Room 4.17
> University of Essex, Colchester CO4 3SQ, UK
> (0044) 01206 872246 / [phone] 6056
> [hidden email]
> website <https://laissue.github.io/>
>
>
> On Wed, 10 Jun 2020 at 15:15, Erik Manders <[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 all,
> >
> > As you may know It has been a while ago that I have worked on methods to
> > reduce the toxic effects of excitation light in fluorescence microscopy.
> > When we worked on Controlled Light Exposure Microscopy (nowadays called
> > Spatially Controlled Illumination Microscopy in order to reduce
> > confusion...) there was not much know about the photo-physical background
> > of phototoxicity. There wasn't even any paper describing dose-effect of
> > light in microscopy.
> >
> > Simple questions like:
> > - How much light kills a cell?
> > - How much light stops the cell cycle without killing the cell?
> > - What is the role of the fluorophore?
> > - Are some fluorophores more phototoxic than others?
> > - Are oxygen radicals directly involved or singlet oxygen?
> > - Has dose-rate (at constant dose) an influence on cell happiness?
> > - A thousand other questions, but the main simple question is: How much
> > light kills a cell???
> >
> > Who has an answer to these questions or knows the title of a paper that
> > goed into this kind of detail. I am out of this subject for some years
> now,
> > but I am very interested to get an update and see if any progress has
> been
> > made since the end 90s.
> >
> > Kind regards, Erik Manders
> >
>

[Anna Ashton]

*****
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 Erik,

There's also a recent paper from Claire Brown's lab which looked at this, including the benefit of using lower intensity power with a longer exposure time to reduce phototoxicity.

https://jcs.biologists.org/content/133/4/jcs242834.long

Best wishes
Anna

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Erik Manders
Sent: 10 June 2020 15:15
To: [hidden email]
Subject: Phototoxicity

*****
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 all,

As you may know It has been a while ago that I have worked on methods to reduce the toxic effects of excitation light in fluorescence microscopy. When we worked on Controlled Light Exposure Microscopy (nowadays called Spatially Controlled Illumination Microscopy in order to reduce confusion...) there was not much know about the photo-physical background of phototoxicity. There wasn't even any paper describing dose-effect of light in microscopy.

Simple questions like:
- How much light kills a cell?
- How much light stops the cell cycle without killing the cell?
- What is the role of the fluorophore?
- Are some fluorophores more phototoxic than others?
- Are oxygen radicals directly involved or singlet oxygen?
- Has dose-rate (at constant dose) an influence on cell happiness?
- A thousand other questions, but the main simple question is: How much light kills a cell???

Who has an answer to these questions or knows the title of a paper that goed into this kind of detail. I am out of this subject for some years now, but I am very interested to get an update and see if any progress has been made since the end 90s.

Kind regards, Erik Manders

[Erik Manders-3]

*****
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.
*****

Thank you all for your reactions on this item. I got quite some tips and references to papers. Thanks for that! It seems that in the last 10 years, quite some decent research has been done on phototoxicity in fluorescence microscopy. At the moment we are exploring how the RCM is performing in life cell imaging (see www.confocal.nl/timelapse). It seems that a more sensitive detector (camera instead of PM tubes), the concentration of light (due to super resolution proporties of RCM) and possibility to open the pinhole a bit more (since you do not loose lateral resolution by doing that in RCM imaging) helps to increase sensitivity a lot and therefore to reduce light dose and therefore to reduce phototoxicity. I'm curious what you think of this and if you think this makes sense.

Kind regards, Erik

[yjf]

*****
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 EriK

   Nikon A1 confocal still have The CLEM function, but it seems very few sold .

    Best wishes
 
       Jiangfeng


           
From: [hidden email] [mailto:[hidden email]] 代表 Erik Manders
Sent: 2020年6月10日 22:15
To: [hidden email]
Subject: Phototoxicity

*****
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 all,

As you may know It has been a while ago that I have worked on methods to reduce the toxic effects of excitation light in fluorescence microscopy. When we worked on Controlled Light Exposure Microscopy (nowadays called Spatially Controlled Illumination Microscopy in order to reduce confusion...) there was not much know about the photo-physical background of phototoxicity. There wasn't even any paper describing dose-effect of light in microscopy.

Simple questions like:
- How much light kills a cell?
- How much light stops the cell cycle without killing the cell?
- What is the role of the fluorophore?
- Are some fluorophores more phototoxic than others?
- Are oxygen radicals directly involved or singlet oxygen?
- Has dose-rate (at constant dose) an influence on cell happiness?
- A thousand other questions, but the main simple question is: How much light kills a cell???

Who has an answer to these questions or knows the title of a paper that goed into this kind of detail. I am out of this subject for some years now, but I am very interested to get an update and see if any progress has been made since the end 90s.

Kind regards, Erik Manders