Posted by
Arne Seitz on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Leica-TCS-SP2-LCS-and-Win7-tp7582853p7582929.html
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/softwareThe 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?
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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
--
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Steffen Dietzel, PD Dr. rer. nat
Ludwig-Maximilians-Universität München
Walter-Brendel-Zentrum für experimentelle Medizin (WBex) Head of light microscopy
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