Re: Numbers for Airy pattern graphs?

Posted by Arne Seitz on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Leica-TCS-SP2-LCS-and-Win7-tp7582853p7582934.html

Dear Steffen,

the PSF lab is "only" calculating the excitation point spread function.
To simulate the effect of the pinhole you needed to modify the emission PSF.

Best
Arne
 

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Steffen Dietzel
Sent: lundi 17 novembre 2014 14:40
To: [hidden email]
Subject: Re: Numbers for Airy pattern graphs?

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Dear Arne,

that does look indeed like an interesting piece of software, e.g. for demonstrating the need to avoid refraction index mismatches. However, although it says it is for confocal microscopy, I do not see an option to play around with pinhole size, so I am afraid it will not work for my specific purpose. The "getting started"-pdf does not contain the words 'pinhole' or 'Airy'. Did I overlook something?

Steffen

Am 17.11.2014 13:29, schrieb Seitz Arne:

> Dear Steffen,
>
> PSFLab is a freely available software with which you can create the excitation point spread function of a confocal microscope.
> http://onemolecule.chem.uwm.edu/index.php/software
>
> The PSF generator can be used in order to calculate a WF point spread function (which can be considered to be similar with the emission PSF of a confocal microscope).
>
> http://bigwww.epfl.ch/algorithms/psfgenerator/
>
> If you are interested in a "simple" airy pattern in the focal plane I can provide you with an ImageJ plugin.
>
> Regards
> Arne
>  
>
> -----Original Message-----
> From: Confocal Microscopy List
> [mailto:[hidden email]] On Behalf Of Steffen Dietzel
> Sent: lundi 17 novembre 2014 11:40
> To: [hidden email]
> Subject: Numbers for Airy pattern graphs?
>
> *****
> 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.
> *****
>
> Dear Confocalists,
>
> for teaching purposes, I would like to have a graph with the following:
>
> 1) Intensity profile through the center of the Airy pattern (x,y) of a fluorescent point for a wavelength of 500 nm (as, say, mean between 488 exc. and 512 em) in widefield, with the distance from the maximum on the x-axis in nano- or micrometers and the normalized intensity on the y-axis, say for a NA 1.4 Objective.
>
> 2) Same for confocal, closed pinhole.
>
> Bonus:
> 3) Same for confocal/two photon with 900 nm excitation
> 4) All of the above along the optical axis instead of the x,y plane.
>
>
> One purpose of this graph (more precisely: comparison of 1 and 2) would be to visualize with typical parameters why in the confocal/two-photon case with the steeper fall in intensity two Airy patterns can be closer together and still have a 26% dip in intensity between them (called extended Rayleigh-criterion, I believe?) although the ring with the first minimum has the same diameter.
>
> I am sure I could find that or a similar version in some book or article, however I would like to include this in material published on the web. And to avoid copyright issues the graphs have to be in the public domain or with a Creative Commons Attribution License (CC-by).
> Which probably means that they have to be made new. I would be perfectly happy to make such graphs myself if I had the respective data (i.e.
> columns with numbers for the x- and Y-values) to generate them in Excel, Calc or Graphpad. However, as a mere biologist I don't know how to generate such columns.
>
> Therefore I would like to ask if someone would be willing to help out and generate such columns with numbers. I assume that it is not that much work for somebody with good insight in Airy pattern formulas and the right program at hands - and hope I am not wrong with that assumption. In particular data for graphs 1 and 2 above would be very helpful.
>
> I am thinking that covering the distance from -5 to +5 µm in 10 nm steps would probably be more than sufficient.
>
> Cheers
>
> Steffen
>
>
> --
> ------------------------------------------------------------
> 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
>

--
------------------------------------------------------------
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