Single-Molecule Tracking dataset for histone H3 (Hht1) in yeast Saccharomyces cerevisiae
This dataset contains raw time-lapse movies acquired for tracking histone H3 (Hht1) in Saccharomyces cerevisiae. The histone H3 (HHT1) was C-terminally tagged with -HaloTag and sparsely labeled with HaloTag ligand (JF646-HTL). To identify the nuclear boundaries, GFP-LacI-NLS (Nuclear Localization Sequence) was expressed. The cells were grown to log phase in SC media, labeled with JF646-HTL, and imaged under the Highly Inclined and Laminated Optical Sheet (HILO) microscope with a pixel size of 117 nm. Time-lapse movies were acquired (with a 647 nm laser) at three different time intervals (15 ms, 200 ms, and 1000 ms). After each time-lapse movie, a single image was captured with a 488 nm laser to generate images for nuclear boundaries. These movies were tracked using three different MATLAB-based software packages (MatlabTrack, TrackIt, and DiaTrack (Sojourner and Spot-On)) to extract various biophysical parameters of histone H3 dynamics, such as mean squared displacement plot, diffusion coefficient, bound fraction, residence time, jump distances and jump angles. This dataset also contains tracked files from all three software packages, so users can use those files as examples to learn how to process the data and what to expect from the tracked files. The histone H3 is generally used as a gold standard for chromatin-bound molecules in live-cell single-molecule imaging. The raw timelapse movies provided can be used with any other tracking software to compare the performance of various tracking algorithms for single-molecule tracking.
Department of Biotechnology, Ministry of Science and Technology, India