Re: Light Lab by Carl Zeiss Microscopy for iOS devices

Posted by Cameron Nowell on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Light-Lab-by-Carl-Zeiss-Microscopy-for-iOS-devices-tp7184469p7194364.html

Ok for some reason it didn’t let me post my first response so here it is again

Hi Steffen,

The SVI website has a Nyquist calculator that does what you want

http://www.svi.nl/NyquistCalculator


It calculates optimal Nyquist for various settings. It can also generate a PSF of a refractive mismatch to show you what goes wrong if you want. It seems to be limited to <10um into your sample though for some reason.

The calculations for resolution at different depths is easy enough to code up for a webpage, but the brightness shift is rather tricky. There are so many parameters that will effect the brightness of your fluorophore as you go deeper into your sample (even in a perfectly matched situation) that I don't think it would be possible to calculate it. You could get a theoretical value but I doubt it would reflect reality in any way in an actual tissue or clump of cells.

If anyone knows of some formulas to do these sort of calculations send em through and I can put a calculator up on my facilities webpage.


Cheers

Cam


Cameron J. Nowell
Microscopy Manager
Centre for Advanced Microscopy
Ludwig Institute for Cancer Research Melbourne - Parkville Branch PO Box 2008 Royal Melbourne Hospital Victoria, 3050 AUSTRALIA
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Fax: [phone]
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-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Cameron Nowell
Sent: Tuesday, 17 January 2012 8:26 AM
To: [hidden email]
Subject: Re: Light Lab by Carl Zeiss Microscopy for iOS devices

One other thing, the calculator asks for the back projected pinhole size. You can calculate that here http://www.svi.nl/BackprojectedPinholeCalculator


Cheers

Cam



-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Steffen Dietzel
Sent: Tuesday, 17 January 2012 12:06 AM
To: [hidden email]
Subject: Re: Light Lab by Carl Zeiss Microscopy for iOS devices

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Well, for my part I wouldn't mind an (Android) app for

- z-resolution, prefereably one that includes 2- and 3-photon excitation and

- resolution and brightness at various depths under refraction index mismatch conditions (e.g. sample embedded in glycerol instead of immersion oil or underneath a coverslip with a dipping objective).

Life isn't always as simple as 0.61 lambda/NA.


Even better though than a bunch of apps for various systems would a calculator running in a web browser based on common standards so that it would be accessible on every system.


Steffen


On 16.01.2012 02:08, Guy Cox wrote:

> There isn't any difference between widefield and confocal resolution in any normal fluorescence mode.   Do you really need an app to work out 0.61 lambda/NA?  And then to divide by 2.3 to get Nyquist?  If you are in multiphoton it's a little more complicated since root 2 (1.414) comes in, but you can just use the formula 0.43 lambda / NA.
>
> Guy
>
> Optical Imaging Techniques in Cell Biology
> by Guy Cox    CRC Press / Taylor&  Francis
>       http://www.guycox.com/optical.htm 
> ______________________________________________
> Guy Cox, MA, DPhil(Oxon), Honorary Associate, Australian Centre for
> Microscopy&  Microanalysis, Madsen Building F09, University of Sydney,
> NSW 2006
>
> Phone [phone]     Fax [phone]
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