2P laser coupled to a DMD?

[Gary Laevsky]

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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.

[Craig Brideau]

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You have two approaches. Find the DMD device with the greatest power
handling that is in your budget, then back-calculate if that is sufficient
for the 'pixel' size you need on your sample for successful activation.
Alternatively, calculate the energy density per unit area per pixel you
need for activation and shop based on that number.

Craig

On Tue, Nov 19, 2019 at 1:34 PM Gary Laevsky <[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 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.
>

[Zdenek Svindrych-2]

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Hi Gary,

it's certainly not easy :-). If you're thinking about turning just few
pixels on that would define the locations of photoactivation (so the DMD is
a mask that is conjugate to the image plane), then, no, this won't work. At
least not over any reasonable field of view. The throughput would be too
low and you won't get enough peak power for two-photon absorption to
manifest.

If you're thinking to encode a diffraction pattern on the DMD that would
steer your beam (single spot in the image plane at a time), this would
probably work, but I don't know any commercial product like this, as
galvo-based steering is more efficient, simpler, cheaper; only quite a bit
slower. There are vis-laser solutions available commercially, but those use
LCOS for better efficiency.

From the power-handling point of view, you won't really damage a DMD or
LCOS SLM with a 1 W average power spread over the full chip. The peak power
won't matter as it's not focused to a spot on the DMD; the 80-or-so MHz
pulses just might disturb the electronics behind the pixels in some very
unfavorable cases.

Best, zdenek

Zdenek Svindrych, Dartmouth College


On Tue, Nov 19, 2019 at 3:34 PM Gary Laevsky <[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 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.
>

--
--
Zdenek Svindrych, Ph.D.
Research Associate - Imaging Specialist
Department of Biochemistry and Cell Biology
Geisel School of Medicine at Dartmouth

[Ian Dobbie]

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Hi Gary

Do beware DMD’s have a very nasty gotcha with coherent illumination. They work brilliantly with an incoherent source but once your source is coherent you have a rather crap blazed grating as each pixel mirror is offset by almost the same amount which produces nasty blaze effects. The wide bandwidth of a pulsed laser is likely to produce nasty angle dependent variability due to the blaze.

Ian

Sent from my iPhone

> On 19 Nov 2019, at 20:34, Gary Laevsky <[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 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.

[Benjamin Smith]

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*****

Are you wanting temporal or spatial non-linearity?  For temporal, it would
be much easier to use a Pockels cell, and you could even drive it using the
analog output on your electrophysiology equipment to get perfect temporal
correlation.  For spatial (as in holographic), as others have said, you
would need an SLM on the focal plane, and likely a second spectral SLM for
temporal encoding to move the probe in the axial domain.  DMDs are actually
pretty terrible at high speed intensity coding because they encode 8-bit
gray scales by sequentially flashing eight binary masks, each representing
one of the eight bits to be encoded.

On Tue, Nov 19, 2019 at 1:52 PM Zdenek Svindrych <[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 Gary,
>
> it's certainly not easy :-). If you're thinking about turning just few
> pixels on that would define the locations of photoactivation (so the DMD is
> a mask that is conjugate to the image plane), then, no, this won't work. At
> least not over any reasonable field of view. The throughput would be too
> low and you won't get enough peak power for two-photon absorption to
> manifest.
>
> If you're thinking to encode a diffraction pattern on the DMD that would
> steer your beam (single spot in the image plane at a time), this would
> probably work, but I don't know any commercial product like this, as
> galvo-based steering is more efficient, simpler, cheaper; only quite a bit
> slower. There are vis-laser solutions available commercially, but those use
> LCOS for better efficiency.
>
> From the power-handling point of view, you won't really damage a DMD or
> LCOS SLM with a 1 W average power spread over the full chip. The peak power
> won't matter as it's not focused to a spot on the DMD; the 80-or-so MHz
> pulses just might disturb the electronics behind the pixels in some very
> unfavorable cases.
>
> Best, zdenek
>
> Zdenek Svindrych, Dartmouth College
>
>
> On Tue, Nov 19, 2019 at 3:34 PM Gary Laevsky <[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 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.
> >
>
>
> --
> --
> Zdenek Svindrych, Ph.D.
> Research Associate - Imaging Specialist
> Department of Biochemistry and Cell Biology
> Geisel School of Medicine at Dartmouth
>

--
Benjamin E. Smith, Ph. D.
Imaging Specialist, Vision Science
University of California, Berkeley
195 Life Sciences Addition
Berkeley, CA  94720-3200
Tel  [phone]
Fax [phone]
e-mail: [hidden email]
https://vision.berkeley.edu/faculty/core-grants-nei/core-grant-microscopic-imaging/

[samuel connell]

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*****

Commercial Reply:

Good Afternoon Gary et al,

3i has produced a Digital Holography system called Phasor for executing
patterned stimulation for optogenetics with 2P lasers for about 8 years now:

https://www.intelligent-imaging.com/optogenetics

As you guessed it might be, 3i's Phasor is SLM based and re-focuses the
available light into the ROI's of interest. Uniquely, these ROI's can
actually be in multiple/different Z planes while one is imaging distal from
the photostimulation. Phasor is informed by a long-time 3i collaboration
with Valentina Emiliani, currently at the Vision Institut in Paris.

Please do let me know if you have any other questions either here or
off-list.

Best Regards,
-
Sam

Samuel Connell
Director of Sales

Intelligent Imaging Innovations (3i)
3509 Ringsby Court
Denver, CO  80216  USA
1-720-437-6926
www.intelligent-imaging.com



On Tue, Nov 19, 2019 at 12:34 PM Gary Laevsky <[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 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.
>

[Christian Wilms]

*****
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*****

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] / [phone]
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.

[Gary Laevsky]

*****
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] / [phone]
>> 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.

[Christian Wilms]

*****
To join, leave or search the confocal microscopy listserv, go to:
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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] / [phone]
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] / [phone]
> >> 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.

[Gary Laevsky]

*****
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.
*****

Thanks all for the advice.  Much appreciated!

On Fri, Nov 22, 2019 at 5:49 AM Christian Wilms <
[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 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] / [phone]
> 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] / [phone]
> > >> 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.
>

--
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 Annual Meeting at Princeton University,
Nov. 1, 2019.