Re: Multiphoton microscope - Field of View

Posted by Benjamin Smith on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Multiphoton-microscope-Field-of-View-tp7586063p7586075.html

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We use the following grid micrometer with a layer of fluorescein and a coverslip: http://www.edmundoptics.com/test-targets/image-analysis-test-targets/multi-grid-standard-stage-micrometer/58607/

The fluorescence reflects off of the mirrored grid nicely, and allows for the width and height to be measured simultaneously.  It is also really helpful for aligning a brightfield system to an MP scan field, including any transformations.

Cheers,

   Ben Smith

On Wed, Nov 30, 2016 at 4:16 PM, Tamara Howard <[hidden email]> wrote:

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If you have a spare etched micrometer slide, you could "paint" the etched lines with the fluorescent marker. We have a very old, etched line slide that someone probably modified to be fluorescent at some time. Swipe the marker (or make a dye paint) over the etched area, let it dry, then carefully wipe the surface clean & at least some of the fluor should stay in the etched lines.

Tamara

...................................................
Tamara Howard
Dept. of Cell Biology & Physiology
University of New Mexico
Albuquerque, NM

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From: Confocal Microscopy List <[hidden email]> on behalf of Gary G. Li <[hidden email]>
Sent: Wednesday, November 30, 2016 4:09 PM
To: [hidden email]
Subject: Re: Multiphoton microscope - Field of View

 

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You can also paint the printed grid with a fluorescent marker, to make it a crude fluorescent target.   

On Wed, Nov 30, 2016 at 5:52 PM, Reece, Jeff (NIH/NIDDK) [E] <[hidden email]> wrote:

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Hi Tim,

 

If you are happy with precision of a few percent, you could use Powerpoint or similar software, and a decent printer, to print out your own crude sample on regular paper, and image it in transmitted or reflected light.  The spacing on my sample goes down to 500 microns, but your printer might do much better.

 

Cheers,

Jeff

 

From: Gary G. Li [mailto:[hidden email]]
Sent: Wednesday, November 30, 2016 3:00 PM
To: [hidden email]
Subject: Re: Multiphoton microscope - Field of View

 

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Look at something with a known size, e.g., a fluorescent slide with known grid spacing.  

 

On Wed, Nov 30, 2016 at 2:01 PM, Tim <[hidden email]> wrote:

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Hi all, sorry if this has been discussed before (which I'm sure it has) but I couldn't find any relevant discussions through search. I was wondering how does one go about measuring field of view on a custom designed multiphoton microscope?

Thanks,
Tim

 

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