Original paper https://nph.onlinelibrary.wiley.com/doi/10.1111/nph.17792.
The program is written to facilitate repetitive calculations of fluorescence recovery after photobleaching (FRAP). For correct work, the program requires python 3.6 and above - https://www.python.org/downloads/.
The program works with max 3 ROIs at once (plus one background and one reference ROIs). The number of samples is unlimited. Names of columns with each ROI data are fixed as follows: col_1(or another number), long_1, and lat_1. Columns with background, reference and time, should be named by the same model: back_1, ref_1, and time_s_1, respectively. Input format should look like following exaple:
|time_s_1|back_1|ref_1 |cp_1 |long_1 |lat_1|time_s_2|back_2| |0 |24.332|161.397|398.839| 173.005|NaN | 0 |23.092| etc....
If less than 3 ROIs per sample are needed, the user can leave the column with the respective name empty.
Output for graphic plot undergoes time conversion. It's necessary because different samples often have a slightly different pace, so mean and SEM cannot be calculated easily. Each measurement is assigned to a 1-sec interval. If for one interval there is more than one measurement - mean of these values will be taken. The conversion allows free comparison and averaging of measurements with any pace below or equal to 1 s. This approximation doesn't influence the curve's shape but can slightly affect separate time points(as the average is calculated). If you don't want to make any time conversion - use an automatically generated data frame for sigmaplot as a result.
Program gets excel tab as an input, extracts columns of interest, normalizes them, normalizes bleached ROIs to non-bleached reference (optional, recommended if undesired bleaching is too strong), subtracts background (optional), transforms timescale, calculates mean, standard error of the mean, and groups output to the new excel tab. Created by Kashkan Ivan. 2019.02.12