... | ... | @@ -30,7 +30,7 @@ Here we explain how to install and use Fluorescence Correlation Spectroscopy (FC |
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#### Procedure
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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.
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> Remember to switch on background indicator in correction steps and then fill offset calculated in step 1
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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.
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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.
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**Important:** do not forget to save fluctuation traces with ACF fitting curves (Export all traces button) as .cof, .itr files.
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3. Prepare the following [structure of files](structure of files).
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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 |
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2. **calibration.csv**
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Here you can find the concentrations of POI calculated using calibration parameters.
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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)
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3. **calibration_plot.png**
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... | ... | @@ -86,3 +86,7 @@ The image of the final calibration plot. |
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4. **calibration_plot.csv**
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The table with data points in calibration plot (can be used to plot the calibration plot with other software)
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5. **map folder**
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The folder contains processed concentration maps in case the concentration maps metanode was used in analysis. |