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Unruh, Jay on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Zeiss-FCS-Analysis-tp7582564p7582565.html
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Hi Eric,
The .fcs output file should contain information at whatever resolution you calculate the correlation at. There is probably an internal setting for that.
I have free software for fcs analysis in ImageJ here:
http://research.stowers.org/imagejplugins/fcs_plugins.html That software requires you to save the .raw files. There should be a setting in the zeiss software for that as well. It recalculates the autocorrelation functions using a slightly different method than the Zeiss software, so you can select the appropriate temporal sampling frequency for your analysis.
Jay
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From: Confocal Microscopy List [mailto:
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Sent: Wednesday, August 27, 2014 10:43 AM
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Subject: Zeiss FCS Analysis
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Hi All,
We are using a Zeiss 710 with a GaAsP detector to perform FCS. We are using a dilute (500nM) solution of Rhodamine B and the free on-board FCS software. We were hoping to use the known diffusion rate and concentration of this molecule to calculate the structural parameter of our FCS volume. When we look at our sample, we can see a reasonable count rate, a correlation of around 1, and a CPM value greater than 1. When we run the experiment, we see a nice autocorrelation curve. However, when we output the data, the best time resolution we can obtain is 1ms. There is clearly higher frequency data that is being used to generate the autocorrelation curves, but we cannot get it. Does anyone have any experience with using the data from a 710 to generate your own autocorrelation curves and do your own analysis? Additionally, if anyone could recommend any free software (Matlab or R) for performing FCS analysis, we would greatly appreciate it. Thanks for your help.
Best regards,
Eric
Centre for Gene Regulation and Expression College of Life Sciences University of Dundee MSI/WTB/JBC Complex Dow Street Dundee DD1 5EH United KIngdom
+44 (0)1382 385118
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