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#### 2. Perform 2 separate FA analysis sessions for POI and FP data:
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- Perform [correlations calculation and correction](https://git.embl.de/grp-ellenberg/FCSAnalyze/-/wikis/Fa_Load_and_Correlate)
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- Do [fitting of correlations with ACF](https://git.embl.de/grp-ellenberg/FCSAnalyze/-/wikis/Fa_fit_fcs)
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- Perform [fitting of correlations with ACF](https://git.embl.de/grp-ellenberg/FCSAnalyze/-/wikis/Fa_fit_fcs)
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<br><be>
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- Save the processed data: during save, export and report step, click FA format to save corresponding res files (**mFP.res** and **POI.res**) and save (Export all traces button) fluctuation traces with ACF fitting curves for POI and FP.
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- Go to save, export and report step, click FA format to save corresponding res files (**mFP.res** and **POI.res**) and save (Export all traces button) fluctuation traces with ACF fitting curves for POI and FP.
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#### 3. Calculate the effective confocal volume:
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- Follow the same steps described in bullet point 2 to obtain **dye.res**
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<be><br>
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**or**
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- If the effective confocal was measured without FA*, the user can specify this volume in fl in the main user input. This volume will be then used to calculate concentrations for the calibration plot. The ways to obtain the effective confocal volume outside the FCSpipelineEMBL_KNIME can be found in technical details.
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* in case there is no dye.res file in your analysis
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#### Important notes regarding FA fitting step:
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- There are **different fitting models** used for dye and FP&POI.
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