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

<p>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.&nbsp; By utilizing high-speed fluorescent microscopy, we tracked a transmembrane protein &ndash; the high-affinity IgE receptor &ndash; and an outer-leaflet protein &ndash; GPI-anchored protein &ndash; 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.&nbsp;&nbsp;</p> <p>The data was classified as &ldquo;Class I&rdquo; and &ldquo;Class II&rdquo; describing two categories of RBL-2H3 cells.&nbsp; In "Class I", the cells were untransfected.&nbsp; In &ldquo;Class II&rdquo;, the cells were transfected to express GFP-GPI-anchored fusion protein. The zip folder names that include &ldquo;IgE Untreated&rdquo; 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 &ldquo;SRImage&rdquo; folder.</p> <p>The data with folder names including with &ldquo;IgE Treated&rdquo; 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.</p> <p>Similarly, the data with folder names starting with &ldquo;GPI Untreated&rdquo; 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.</p> <p>The data with folder names starting with &ldquo;GPI Treated&rdquo; 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.</p> <p>The number within each folder name represents an individual experiment and the corresponding data collected under the same conditions.</p> <p>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.&nbsp;</p> <p>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.&nbsp;</p> <p>&nbsp;</p> <p>&nbsp;</p> <p>&nbsp;</p>