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There may be a bug of some sort in your Zeiss installation. There may also be a setting under the maintain menu to keep the raw trajectories. Our LSM 780 exports the raw photon mode trajectories just fine as .raw files. You have to parse the binary data, but it is certainly there for detailed analysis.
Jay
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Philipp Steffen
Sent: Wednesday, August 27, 2014 11:06 AM
To: [log in to unmask]
Subject: Re: Zeiss FCS Analysis
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Dear Eric,
As far as I know the Zeiss software does not allow you to export the raw data. My understanding is that the ACF is generated "online" and that the user can only export fluctuation traces which are binned in certain time intervals. This seems to be a limitation of the software if you want to generate your own ACF.
However if you only want to use custom software (e.g. Matlab) to fit the ACF to a certain model you can of course work using the ACF exported from the Zeiss software.
Best regards,
Philipp
Philipp A. Steffen, PhD
Vienna
(Previously at IMBA,
Ringrose lab)
> Am 27.08.2014 um 17:42 schrieb Eric Griffis <[log in to unmask]>:
>
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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
>
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