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howto:calculate_fccs_trace_with_the_grouped_fcs_script [2014/05/23 07:44] – adminhowto:calculate_fccs_trace_with_the_grouped_fcs_script [2015/02/25 14:01] – bornemann
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 {{tag> analysis howto tutorial FCS FCCS Cross-Correlation  SymPhoTime}} {{tag> analysis howto tutorial FCS FCCS Cross-Correlation  SymPhoTime}}
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-====== Calculate FCCS Traces with the Grouped FCS Script ======+====== Calculate Multiple FCCS Traces with the Grouped FCS Script ======
  
 ===== Summary ===== ===== Summary =====
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 This tutorial shows step-by-step, how to calculate a spectral cross-correlation curve and the corresponding autocorrelation curves of a dye-labeled DNA-oligonucleotide using the Grouped FCS script. This tutorial shows step-by-step, how to calculate a spectral cross-correlation curve and the corresponding autocorrelation curves of a dye-labeled DNA-oligonucleotide using the Grouped FCS script.
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 ===== Background Information ===== ===== Background Information =====
  
-**Note:** Fluorescence Cross-Correlation Spectroscopy (FCCS) is a method to determine molecular interactions between two molecules labeled with different dyes. Usually, the dyes attached to the two molecules of interest differ in their absorption and emission properties. They can also vary in their fluorescence lifetime, for more information please check the tutorials for Fluorescence Lifetime Correlation Spectroscopy (FLCS).\\+Fluorescence Cross-Correlation Spectroscopy (FCCS) is a method to determine molecular interactions between two molecules labeled with different dyes. Usually, the dyes attached to the two molecules of interest differ in their absorption and emission properties. They can also vary in their fluorescence lifetime, for more information please check the tutorials for Fluorescence Lifetime Correlation Spectroscopy (FLCS).\\
 To quantify the cross-correlation signal and connect it to a binding fraction, a calibration of the effective confocal volumes for each dye and the overlapping confocal volume must be performed.\\ To quantify the cross-correlation signal and connect it to a binding fraction, a calibration of the effective confocal volumes for each dye and the overlapping confocal volume must be performed.\\
 The calibration can be done using a molecule labeled with both fluorophores.\\ The calibration can be done using a molecule labeled with both fluorophores.\\
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 **Note:** In a sample as measured here by a combination of pulsed and cw excitation, it is possible to remove the false positive cross-correlation, which significantly simplifies the binding analysis. This is explained in the tutorial [[howto:using_the_flcs_script_for_spectral_crosstalk_removal_via_flccs|Using the FLCS script for spectral crosstalk removal via FLCCS]]. **Note:** In a sample as measured here by a combination of pulsed and cw excitation, it is possible to remove the false positive cross-correlation, which significantly simplifies the binding analysis. This is explained in the tutorial [[howto:using_the_flcs_script_for_spectral_crosstalk_removal_via_flccs|Using the FLCS script for spectral crosstalk removal via FLCCS]].
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howto/calculate_fccs_trace_with_the_grouped_fcs_script.txt · Last modified: 2020/04/01 15:05 by buschmann