@@ -30,7 +30,7 @@ Here we explain how to install and use Fluorescence Correlation Spectroscopy (FC
#### Procedure
1. Process a WT data with FA to obtain WT.res. A detailed explanation of FA analysis procedure can be found [here](https://git.embl.de/grp-ellenberg/FCSAnalyze/-/wikis/home). Only import, correction, and export steps of FA session are needed. Fill a WT user input and execute Python Source node. Use returned offset value for correction steps in further FA sessions for FP and POI.
> Remember to switch on background indicator in correction steps and then fill offset calculated in step 1
2. Perform three separate sessions of FA analysis: for FP, for POI, and for dye. Do all intermediate FA steps (correlations calculation, correction, fitting of correlations with ACF) to obtain mFP.res, POI.res, and dye.res.
2. Perform three separate sessions of FA analysis: for FP, for POI, and for dye as described [here](https://git.embl.de/grp-ellenberg/FCSAnalyze/-/wikis/home). Do **all** intermediate FA steps (correlations calculation, correction, fitting of correlations with ACF) to obtain mFP.res, POI.res, and dye.res.
**Important:** do not forget to save fluctuation traces with ACF fitting curves (Export all traces button) as .cof, .itr files.
3. Prepare the following [structure of files](structure of files).
4. Specify parameters in the main user input
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@@ -77,7 +77,7 @@ info.csv is generated inside the main user input. This is the main output with a
2.**calibration.csv**
Here you can find the concentrations of POI calculated using calibration parameters.
Here you can find the concentrations of POI in corresponding FCS positions. You can find the explanations of how concentrations are calculated on [technical details](technical details)
3.**calibration_plot.png**
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@@ -86,3 +86,7 @@ The image of the final calibration plot.
4.**calibration_plot.csv**
The table with data points in calibration plot (can be used to plot the calibration plot with other software)
5.**map folder**
The folder contains processed concentration maps in case the concentration maps metanode was used in analysis.
