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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
<[log in to unmask]> wrote:
> *****
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> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> 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
>>> <[log in to unmask]> 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 <[log in to unmask]> 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
>>>>
>>>> [log in to unmask]
>>>
>
>
>
> 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
>
> [log in to unmask]