Re: recommended distortion target for multiphoton?

Posted by Michael Giacomelli on
URL: http://confocal-microscopy-list.275.s1.nabble.com/recommended-distortion-target-for-multiphoton-tp7582387p7582663.html

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Just to follow up, I contacted MicronLaser.com and got several 1 cm^2
grid target etched into borosilicate glass.  They are ~20 um deep and
so I can flow different fluorophores through.  The cost was very
reasonable, but they were machined with a CO2 laser, and so have some
thermal microcracking that makes them unsuitable as resolution
targets.  Still, they are virtually indestructable (can park the beam
forever at one point and you'll never damage it), and the very large
size makes them quite handy for testing translation stages and
mosaicing software.

I'd still like to find some who can do a nanosecond pulsed laser etch
and see about getting custom resolution targets cut, but overall, the
micronlaser parts were quite reasonable for what I needed.

Mike

On Wed, Jul 23, 2014 at 2:00 PM, Craig Brideau <[hidden email]> wrote:

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>
> I've been bugging various companies for years to get a good 2P and confocal
> fluoresence target developed, but thus far nobody's come up with anything
> that really works that costs less than $1000.
>
> Craig Brideau
>
>
> On Wed, Jul 23, 2014 at 11:26 AM, Michael Giacomelli <[hidden email]> wrote:
>
>> *****
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>>
>> Unfortunately, it is extremely difficult to record the reflection mode
>> image in our microscope, as the NIR light will not pass through the
>> dichroic, and even if we changed out the dichroic for a beam splitter,
>> out PMTs are not sensitive to NIR light.
>>
>> Translating a bead though is a very interesting idea.  We use a high
>> precision thorlabs MLS stage.  I will double check the repeatability,
>> but in theory I could program it to perform an extremely high
>> resolution distortion measurement.  Of course, getting a single bead
>> onto a coverslip (or else programming software to automatically
>> identify the same bead across hundreds of frames) will be somewhat
>> tricky.
>>
>> Its a shame that getting targets for multiphoton is so difficult.
>> Ideal I suppose would be something etched in glass so that I could
>> flow my own fluorophore through it.  I will have to look around and
>> see if I can get a lithography facility to make something like that.
>>
>> Thanks,
>> Mike
>>
>> On Wed, Jul 23, 2014 at 3:19 AM, Mark Cannell
>> <[hidden email]> wrote:
>> > *****
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>> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
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>> posting.
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>> >
>> > Thinking about this a bit more, I realised that your existing grid is
>> perfectly suitable as you don’t need 2P excitation to measure field
>> distortion at the wavelength you are using, Just turn the power down a lot
>> and record a reflection image,
>> >
>> > Cheers Mark
>> >
>> >>>
>> >>> Hi Mark,
>> >>>
>> >>> I'm assuming you mean beads deposited in a grid or something similar?
>> >>> Could you point me to where you found this?
>> >>>
>> >>> Thanks,
>> >>> Mike
>> >>>
>> >>> On Tue, Jul 22, 2014 at 3:58 AM, Mark Cannell
>> >>> <[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 Michael
>> >>>>
>> >>>> I’ve always used beads to test the microscope. They are relatively
>> cheap
>> >>> and if you blow one up it don’t matter. As you say, evaporated metal
>> >>> targets are too easily damaged (as Jim Pauley and I discovered when
>> testing
>> >>> my microscope back in ’95!)
>> >>>>
>> >>>> HTH
>> >>>>
>> >>>> Mark
>> >>>>
>> >>>> On 22/07/2014, at 4:30 am, Michael Giacomelli <[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.
>> >>>>> *****
>> >>>>>
>> >>>>> Could someone recommend a good multiphoton distortion target?
>> >>>>> Preferably one that is relatively robust against damage.
>> >>>>>
>> >>>>> Currently I am using a thorlabs wire grid target with fluorophore
>> >>>>> behind it.  The beam is attenuated by the metal, resulting in an
>> image
>> >>>>> of the grid.  However, passing through the entire 1 mm slide greatly
>> >>>>> reduces my resolution, and unless I am extremely careful, the grid is
>> >>>>> ablated by the beam.
>> >>>>>
>> >>>>> Thanks,
>> >>>>> Michael
>> >>>>
>> >>>>
>> >>>> Mark  B. Cannell Ph.D. FRSNZ
>> >>>> Professor of Cardiac Cell Biology
>> >>>> School of Physiology &  Pharmacology
>> >>>> Medical Sciences Building
>> >>>> University of Bristol
>> >>>> Bristol
>> >>>> BS8 1TD UK
>> >>>>
>> >>>> [hidden email]
>> >>>
>> >
>> >
>> >
>> > Mark  B. Cannell Ph.D. FRSNZ
>> > Professor of Cardiac Cell Biology
>> > School of Physiology &  Pharmacology
>> > Medical Sciences Building
>> > University of Bristol
>> > Bristol
>> > BS8 1TD UK
>> >
>> > [hidden email]
>>