Posted by
Sripad Ram on
URL: http://confocal-microscopy-list.275.s1.nabble.com/discussion-is-open-continues-about-localization-and-super-resolution-tp7581644p7581646.html
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy*****
Hi Chris,
The z-localization precision for astigmatism based 3D localization was worked out by holtzer et al., applied physics letters, 2007, 90, 053902.
On a related note, the precise calculations for the X/Y localization precision that includes background and pixilation was published in 2004 by Raimund Ober group (Ober et al; Biophysical Journal, 2004, 86:1185-1200). You can download the software to calculate the localization precision from here (
http://www.wardoberlab.com/software/fandplimittool/).
Hope this helps.
Regards,
Sripad
-----Original Message-----
From: Confocal Microscopy List [mailto:
[hidden email]] On Behalf Of Christophe Leterrier
Sent: Tuesday, March 04, 2014 3:16 PM
To:
[hidden email]
Subject: Re: discussion is open...continues ... about localization and super resolution
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy*****
This is a great introduction to a question a wanted to ask to the SMLM specialists (I did not find the answer in the above reference). I also posted it on the Nanoscopy LinkedIn group, so I'm sorry for people that will get it two times!
There are several formulas to estimate the lateral (XY) precision of individual localization events in PALM/STORM microscopy. This goes from the crude sigma(PSF)/√Nphotons to more elaborate formulas that includes pixelation and background (such as Thomson & Webb Biophys J 2002, corrected in Mortensen Nat Methods 2010). What I'm looking for is an equation that allows to estimate the precision for the Z localization when using astigmatism-based 3D PALM/STORM. The reference I have found where this precision is studied theoretically (such as Badierostami Applied Phys Lett 2010, Shaevitz Int J Optics 2009) do not use a directly calculable formula.
I assume the precision depends on the photon number and the point on the Z calibration curve with the respective X and Y PSF width. Is anyone aware of such a formula?
Best Regards,
Christophe
2014-03-04 21:52 GMT+01:00 Alberto Diaspro <
[hidden email]>:
> *****
> To join, leave or search the confocal microscopy listserv, go to:
>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> *****
>
>
http://www.nature.com/nmeth/journal/v11/n3/full/nmeth.2843.html>