Re: the use of adaptive optics in microscopy

[Jerry (Gerald) Sedgewick]

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I was at the American Society for Cell Biology earlier this month and
came across a company using adaptive optics with deformable mirrors for
microscopy.  Their setup included a way in which the sample could be
monitored for changes in the index of refraction as the focal plane goes
deeper into the tissue, and then the beam shape is changed via the
deformable mirror.  The net result is greater penetration into the
sample, and I would suspect that this method would be an improvement
over pulse compression.  In my experience, the diameter of the beam at
the back aperture of the lens, particularly when it underfills, vastly
improves penetration into the sample.  This may sacrifice the 2 photon
effect, but it's a good solution when depth is more important than
z-resolution.

The adaptive optic technology is coincidentally very promising in the
world of ophthalmic imaging because the refractive index of the eye
changes from one person to another.

I would be very interested in getting responses from this community
about this technology.  I don't have a commercial interest at all in the
company that is providing adaptive optics for microscopy.

Cheers,

Jerry Sedgewick
--

Jerry Sedgewick
Sedgewick Initiatives
[hidden email]
http://www.imagingandanalysis.com

Author of: "Scientific Imaging with Photoshop: Methods, Measurement, and
Output"

[Craig Brideau]

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Higher NA is more vulnerable to spherical aberration.  As you under fill the
lens, you reduce the NA.  This increases your laser spot size, costing you
some resolution, but in turn the lower NA beam is less vulnerable to
spherical aberration so you get a bit better penetration.

Craig


On Wed, Dec 22, 2010 at 3:44 PM, Jerry (Gerald) Sedgewick <
[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> I was at the American Society for Cell Biology earlier this month and came
> across a company using adaptive optics with deformable mirrors for
> microscopy.  Their setup included a way in which the sample could be
> monitored for changes in the index of refraction as the focal plane goes
> deeper into the tissue, and then the beam shape is changed via the
> deformable mirror.  The net result is greater penetration into the sample,
> and I would suspect that this method would be an improvement over pulse
> compression.  In my experience, the diameter of the beam at the back
> aperture of the lens, particularly when it underfills, vastly improves
> penetration into the sample.  This may sacrifice the 2 photon effect, but
> it's a good solution when depth is more important than z-resolution.
>
> The adaptive optic technology is coincidentally very promising in the world
> of ophthalmic imaging because the refractive index of the eye changes from
> one person to another.
>
> I would be very interested in getting responses from this community about
> this technology.  I don't have a commercial interest at all in the company
> that is providing adaptive optics for microscopy.
>
> Cheers,
>
> Jerry Sedgewick
> --
>
> Jerry Sedgewick
> Sedgewick Initiatives
> [hidden email]
> http://www.imagingandanalysis.com
>
> Author of: "Scientific Imaging with Photoshop: Methods, Measurement, and
> Output"
>

[Paul Herzmark]

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*Austin* Roorda at UC Berkeley does adaptive optics in eyes. Maybe he has an
opinion on it.

I assume part of the problem with Adaptive Optics is that you have to image
to find out what the wavefront characteristics are, then correct them and
image again. So it is slow and bleachy.

Paul Herzmark
Specialist
[hidden email]

Department of Molecular and Cell Biology
479 Life Science Addition
University of California, Berkeley
Berkeley, CA  94720-3200
[phone]
[phone] fax


On Wed, Dec 22, 2010 at 2:44 PM, Jerry (Gerald) Sedgewick <
[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> I was at the American Society for Cell Biology earlier this month and came
> across a company using adaptive optics with deformable mirrors for
> microscopy.  Their setup included a way in which the sample could be
> monitored for changes in the index of refraction as the focal plane goes
> deeper into the tissue, and then the beam shape is changed via the
> deformable mirror.  The net result is greater penetration into the sample,
> and I would suspect that this method would be an improvement over pulse
> compression.  In my experience, the diameter of the beam at the back
> aperture of the lens, particularly when it underfills, vastly improves
> penetration into the sample.  This may sacrifice the 2 photon effect, but
> it's a good solution when depth is more important than z-resolution.
>
> The adaptive optic technology is coincidentally very promising in the world
> of ophthalmic imaging because the refractive index of the eye changes from
> one person to another.
>
> I would be very interested in getting responses from this community about
> this technology.  I don't have a commercial interest at all in the company
> that is providing adaptive optics for microscopy.
>
> Cheers,
>
> Jerry Sedgewick
> --
>
> Jerry Sedgewick
> Sedgewick Initiatives
> [hidden email]
> http://www.imagingandanalysis.com
>
> Author of: "Scientific Imaging with Photoshop: Methods, Measurement, and
> Output"
>

[Philippe clemenceau]

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About adaptive optics :

My company was the one exhibiting at ASCB and this year ( 2011) , we are
coming out with a brand new plug and play box which will be adaptable to
commercial microscopes.

To answer to some of the comments I saw :

-If you are using a deformable mirror "alone" to improve the image, it might
take a lot of images ( 100) to get to the optimal "corrected position".

-However if you use a wavefront sensor and work with beads or quantum dots
placed in your sample, you can correct the wavefront immediately, with 1
iteration.

Now, in 3D PALM and STORM techniques, we have demonstrated huge improvements
with adaptive optics, but not just for aberration correction. In regular 3D
palm or STORM, a cylindrical lens is used to do the correspondence between z
depth and the shape of the PSF of the fluorophores. By using the deformable
mirror instead of the cylindrical lens, adaptive optics provides an ideal
programmable lens which helps correcting aberrations, adjust the amount of
astigmatism, therefore adjusting the depth of field and control the z
sensitivity of the 3 D analysis  performed. My company acquired very fresh
results on this in the last few weeks. PALM and STORM users have found this
very useful for their analysis.
People who are interested in this subject should let me know, we are
preparing an application note that will be communicated in the next few weeks.

Philippe Clemenceau
Imagine Optic/Axiom Optics
1 Broadway, 14th floor
Cambridge, MA 02140
[hidden email]