http://confocal-microscopy-list.275.s1.nabble.com/2P-laser-coupled-to-a-DMD-tp7590172p7590217.html
Thanks all for the advice. Much appreciated!
> *****
> 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 Gary,
>
> The power requirement is more relevant if you want to hit multiple cells
> at a time. In general, an SLM-based approach is lossy, as you are working
> with the first order, while the majority of light will end up in the
> non-diffracted zero-order. I know users of commercial solutions who are
> firing 20 W of 1030 nm light into the microscope and getting 500 mW out of
> the objective.
>
> Many of the red-shifted opsins will be efficiently excited by 1030 - 1064
> nm. A wavelength range that has a wide selection of affordable lasers is
> available. If you have an Insight, I would try that first, though.
>
> 1P vs 2P: it will depend heavily on your specimen if 100 microns is
> achievable.
>
> Best of luck, Christian
>
> Dr. Christian Wilms / Research & Development Manager
>
[hidden email] / +44 (0)1825 749933
> www.scientifica.uk.com
>
> Take a look at our NeuroWire blog to see our latest news, guides, videos
> and more.
>
> > -----Original Message-----
> > From: Gary Laevsky <
[hidden email]>
> > Sent: 20 November 2019 14:06
> > Subject: Re: 2P laser coupled to a DMD?
> >
> > *****
> > 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!
> > >
> > > From what I understand, this is totally doable. I'll need to use an
> > > SLM as opposed to a DMD.
> > >
> > > Clarification and additional questions;
> > >
> > > This is for spatial non-linearity. I want to light up "a" cell in a
> > > biofilm/embryo/tissue, at "depth," less than 100 um. I'm not going to
> > > be able to do this with 1P, right? So this WILL limit the selection
> > > of opsins. If I get something like an InSight, 1300 should get me to
> > > the red-shifted? I just emailed Darcy:)
> > >
> > > I'm surprised about the power requirement. I would have thought low
> > > power would be sufficient?
> > >
> > > I'm thinking I have two options; put the SLM between a scan head and
> > > the scope, with divergent then collimating optics to the SLM, and park
> > > the beam of the scan head on the SLM? Or, skip the scan head and
> > > couple the laser/SLM directly to stand. The former will give me
> > > additional capability, but I'm not sure if that light path is viable?
> > >
> > > Thank you ALL very much!
> > >
> > >
> > >
> > > On Wed, Nov 20, 2019 at 4:57 AM Christian Wilms <
> > >
[hidden email]> wrote:
> > >
> > >> Hi Gary,
> > >>
> > >> As others have already said, a DMD won't easily get you the power
> > >> levels that you require for 2P excitation of most optogenetic tools.
> > >> LCoS SLMs are the tool of choice for this. Many commercial suppliers
> > >> of multiphoton microscopes have licensed the technology and now sell
> > >> such add-ons (or have them in development), so it might be worth
> > >> asking the manufacturer of your system.
> > >>
> > >> If you are more the DIY type, have a look at the work by Volodymyr
> > >> Nikolenko and Darcy Peterka from Rafael Yuste's lab at Columbia. This
> > >> paper might be a good introduction:
> > >>
https://doi.org/10.3389/neuro.04.005.2008> > >>
> > >> Darcy and his colleagues (among many others) are really helping push
> > >> this technology and are just a couple of hours from you and are the
> > >> holders of the main patent covering this technology.
> > >>
> > >> In addition to the actual SLM, if you want to target more than a few
> > >> spots simultaneously, you should also be looking into suitable
> > >> lasers, as the ones commonly used in multiphoton imaging tend not to
> > >> hit the required parameters for most opsins.
> > >>
> > >> Best, Christian
> > >>
> > >> Dr. Christian Wilms / Research & Development Manager
> > >>
[hidden email] / +44 (0)1825 749933
> > >> www.scientifica.uk.com
> > >>
> > >> Take a look at our NeuroWire blog to see our latest news, guides,
> > >> videos and more.
> > >>
> > >> > -----Original Message-----
> > >> > From: Gary Laevsky <
[hidden email]>
> > >> > Sent: 19 November 2019 20:33
> > >> > Subject: 2P laser coupled to a DMD?
> > >> >
> > >> > *****
> > >> > 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 All,
> > >> >
> > >> > I THINK this can be done? DMD that can handle high power and
> > >> appropriate
> > >> > reflective characteristics for the wavelength. Or could I do this
> > >> > with
> > >> an SLM?
> > >> >
> > >> > We're looking to do non-linear optogenetic switching.
> > >> >
> > >> > If this is simple, please be gentle. I'm a biologist ...:)
> > >> >
> > >> > If it is simple, can you please direct me to the appropriate vendor.
> > >> >
> > >> > Thanks in advance.
> > >> >
> > >> > --
> > >> > Best,
> > >> >
> > >> > Gary Laevsky, Ph.D.
> > >> > Director, Confocal Imaging Facility Nikon Center of Excellence
> > >> > Co-Founder, North Atlantic Microscopy Society (NAMS)
> > >> >
https://namsmicroscopy.com/ Dept. of Molecular Biology Washington
> Rd.
> > >> > Princeton University
> > >> > Princeton, New Jersey, 08544-1014
> > >> > (O) 609 258 5432
> > >> > (C) 508 507 1310
> > >> >
> > >> > North Atlantic Microscopy Society Spring Meeting at UPENN, April
> > >> > 23,
> > >> 2020.
> > >>
> > >
> > >
> > > --
> > > Best,
> > >
> > > Gary Laevsky, Ph.D.
> > > Director, Confocal Imaging Facility
> > > Nikon Center of Excellence
> > > Co-Founder, North Atlantic Microscopy Society (NAMS)
> > >
https://namsmicroscopy.com/ Dept. of Molecular Biology Washington Rd.
> > > Princeton University
> > > Princeton, New Jersey, 08544-1014
> > > (O) 609 258 5432
> > > (C) 508 507 1310
> > >
> > > North Atlantic Microscopy Society Spring Meeting at UPENN, April 23,
> 2020.
> > >
> >
> >
> > --
> > Best,
> >
> > Gary Laevsky, Ph.D.
> > Director, Confocal Imaging Facility
> > Nikon Center of Excellence
> > Co-Founder, North Atlantic Microscopy Society (NAMS)
> >
https://namsmicroscopy.com/ Dept. of Molecular Biology Washington Rd.
> > Princeton University
> > Princeton, New Jersey, 08544-1014
> > (O) 609 258 5432
> > (C) 508 507 1310
> >
> > North Atlantic Microscopy Society Spring Meeting at UPENN, April 23,
> 2020.
>
Dept. of Molecular Biology
Nov. 1, 2019.