Abstract
Single-Molecule Tracking (SMT) is a powerful method to quantify protein dynamics in live cells. Recently, we have established a data analysis pipeline for estimating various biophysical parameters (mean squared displacement, diffusion coefficient, bound fraction, residence time, jump distances, jump angles, and track statistics) from the single-molecule time-lapse movies acquired from yeast Saccharomyces cerevisiae. We acquired the time-lapse movies using different time intervals (i.e. 15 ms, 200 ms, and 1000 ms) to extract the diffusion parameters (from 15 ms time interval movies) and residence time (from 200 ms and 1000 ms time interval movies). We tracked the single molecules from these movies using three MATLAB-based software packages (MatlabTrack, TrackIT, DiaTrack (Sojourner, and Spot-On)) to quantify various biophysical parameters. In this article, we have quantified the biophysical parameters of chromatin-bound histone H3 (Hht1), labeled using JF646 HaloTag Ligand (HTL), and shared a few raw time-lapse SMT movies for the same. Histone H3 is a chromatin-bound protein and it serves as a benchmark for the stably bound molecules for the SMT experiments. Hence, this dataset can be used by various researchers to quantify the biophysical parameters of chromatin-bound molecules (Histone H3). Any newly developed tracking software can use this dataset to validate the accuracy of its tracking algorithms.