BTCCD is a GUI-based software for the Brightness-gated Two Color Coincidence Detection (BTCCD) analysis of single molecule data from a confocal microscope.

The BTCCD software requires Matlab license (2015b or newer).

1. Download BTCCD software from https://github.com/BTCCD/BTCCD;
2. Unpack the files;
3. Start Matlab and make sure that BTCCD routines are visible for Matlab;
4. Call "TCCD" command in the command window.

Initially TTTR data acquisition format was converted to ASCII files. Input data is file(s) is an ASCII code where each line represents a photon. Columns represent 1st column: Channel number; 2nd column: Macro time (in ns); 3rd column: Micro time (in number of time bins since last laser excitation).

Sample data include single-molecule measurement of double-stranded double-labeled DNA of 48 base pairs, 5'-modified with Alexa488 and 3'-modified with Atto647N. This is a reference sample for BTCCD close to 100% coincidence presented in the publication Höfig H., et al. (2019) Brightness-Gated Two-Color Coincidence Detection Unravels Two Distinct Mechanisms in Bacterial Protein Translation Initiation (manuscript submitted for publication).

Run time of the sample data can take up to several minutes. Coincidence curve calculation time depends on the discretization step that can be changed in the software

For detailed description of the BTCCD method refer to the publication Höfig H., et al. (2019) Brightness-Gated Two-Color Coincidence Detection Unravels Two Distinct Mechanisms in Bacterial Protein Translation Initiation (manuscript submitted for publication).

To report bugs of suggestions use the "Issues" function on GitHub.