http://confocal-microscopy-list.275.s1.nabble.com/Light-Lab-by-Carl-Zeiss-Microscopy-for-iOS-devices-tp7184469p7195785.html
Huygens Professional with a (test)license. You also load your image and
the green button. Your PSF will be visualized in the main window and you
graph appear in the other window with the intensities. You can enlarge
refractive mismatch. Huygens takes that fully into account by measuring
the different PSF's also much deeper in the sample. For multi-photon
data that depth is up to hundreds of microns. So the 10 micron limit in
servers. Feel free to try.
> *****
> To join, leave or search the confocal microscopy listserv, go to:
>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> *****
>
> Cameron,
>
> thank you for sharing this link, a nice tool indeed.
>
> Maybe I didn't have enough coffee yet, but it appears that although
> the PSF is visualized and Nyquist is calculatet, the FWHM (or
> 'resolution') is not given.
>
> A pitty, in particular since their Nyquist values multiplied by 2.3
> and what I have calculated don't really match (in multi-photon examples).
>
> But I don't have the time right now to check which formulas they have
> applied.
>
> Steffen
>
>
> On 16.01.2012 22:57, Cameron Nowell wrote:
>> 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
>> Office: +61 3 9341 3158
>> Mobile: +61 422882700
>> Fax: +61 3 9341 3104
>> Facility Website
>> Linked In Profile
>>
>>
>>
>> -----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
>>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy>> *****
>>
>> 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 +61 2 9351 3176 Fax +61 2 9351 7682
>>> Mobile 0413 281 861
>>> ______________________________________________
>>>
http://www.guycox.net>>>
>>
>>
>> --
>> ------------------------------------------------------------
>> Steffen Dietzel, PD Dr. rer. nat
>> Ludwig-Maximilians-Universität München
>> Walter-Brendel-Zentrum für experimentelle Medizin (WBex) Head of
>> light microscopy
>>
>> Mail room:
>> Marchioninistr. 15, D-81377 München
>>
>> Building location:
>> Marchioninistr. 27, München-Großhadern
>
>