Data Repository: Fast Single-Particle Tracking of Membrane Proteins Combined with Super-Resolution Imaging of Actin Nanodomains

This repository contains a collection of correlated 2D super-resolution images obtained through single molecule localization microscopy and 3D time series of single particle tracking data of membrane proteins.  By utilizing high-speed fluorescent microscopy, we tracked a transmembrane protein – the high-affinity IgE receptor – and an outer-leaflet protein – GPI-anchored protein – at the frame rate of 490 Hz. Subsequently, actin structures of the same cells were captured using a super-resolution microscopy (dSTORM technique). Additionally, this dataset includes Technical Validation to support the transition from live-cell imaging to fixed-cell imaging when adding fixation buffers.  

The data was classified as “Class I” and “Class II” describing two categories of RBL-2H3 cells.  In "Class I", the cells were untransfected.  In “Class II”, the cells were transfected to express GFP-GPI-anchored fusion protein. The zip folder names that include “IgE Untreated” include image time series of fluorescently labeled IgE receptors in untreated RBL-2H3 cells and image series of corresponding super-resolution imaging of fluorescently labeled actin filaments of the same cell in “SRImage” folder.

The data with folder names including with “IgE Treated” are image time series of fluorescently labeled IgE receptors in either phalloidin- or PMA-treated RBL-2H3 cells and corresponding image series of super-resolution imaging of fluorescently labeled actin filaments in the same cell.

Similarly, the data with folder names starting with “GPI Untreated” are image time series of fluorescently labeled GPI-anchored proteins in untreated RBL-2H3 cells and corresponding image series of super-resolution imaging of fluorescently labeled actin filaments in the same cell.

The data with folder names starting with “GPI Treated” are image time series of fluorescently labeled GPI-anchored proteins in phalloidin treated RBL-2H3 cells and corresponding image series of super-resolution imaging of fluorescently labeled actin filaments in the same cell.

The number within each folder name represents an individual experiment and the corresponding data collected under the same conditions.

For all experiments an IR movie was added to monitor the cell morphology during live-cell image and adding initial fixation buffer and it was saved in the tracking file. 

The HDF5 files of all data are available in the second version of this repository. For each sample, there are two files: "Tracking.h5" and "SuperResolution_actin.h5". The "Tracking" files include two groups: single-particle tracking data and IR images during tracking. The "SuperResolution_actin" files contain two groups: super-resolution images of actin filaments and IR reference images for image registeration and drift correction during data collection.