A5004786407- Advanced Fluorescence Microscopy Techniques
- Cell Image Analysis Techniques
- Cell Adhesion Molecules Research
- Monoclonal and Polyclonal Antibodies Research
- Microtubule and mitosis dynamics
- Lipid Membrane Structure and Behavior
- Genomics and Chromatin Dynamics
- RNA Research and Splicing
- Cellular Mechanics and Interactions
- Receptor Mechanisms and Signaling
- Angiogenesis and VEGF in Cancer
- Nuclear Structure and Function
- Glycosylation and Glycoproteins Research
- Photosynthetic Processes and Mechanisms
- Gene Regulatory Network Analysis
- Erythrocyte Function and Pathophysiology
- Cellular transport and secretion
- Caveolin-1 and cellular processes
- Diffusion and Search Dynamics
- Advanced Electron Microscopy Techniques and Applications
- Protein Structure and Dynamics
- Single-cell and spatial transcriptomics
- Biotin and Related Studies
- Skin and Cellular Biology Research
- Microbial Metabolic Engineering and Bioproduction
The University of Texas Southwestern Medical Center
2015-2024
Southwestern Medical Center
2015-2024
Howard Hughes Medical Institute
2021
Tokyo Institute of Technology
2021
University of Zurich
2021
Janelia Research Campus
2021
The University of Texas at Dallas
2018-2020
Harvard University
2007-2013
Center for Systems Biology
2010-2013
Scripps Research Institute
2003-2011
Total internal reflection fluorescence microscopy (TIR-FM) has become a powerful tool for studying clathrin-mediated endocytosis. However, due to difficulties in tracking and quantifying their heterogeneous dynamic behavior, detailed analyses have been restricted limited number of selected clathrin-coated pits (CCPs). To identify intermediates the formation vesicles factors that regulate progression through these stages, we used particle-tracking software statistical methods establish an...
The nuclear lamina is a key structural element of the metazoan nucleus. However, organization major proteins composing poorly defined. Using three-dimensional structured illumination microscopy and computational image analysis, we characterized supramolecular structures lamin A, C, B1, B2 in mouse embryo fibroblast nuclei. Each isoform forms distinct fiber meshwork, with comparable physical characteristics respect to mesh edge length, face area shape, connectivity form faces. Some...
Binding of ligands by immunoreceptors is thought to be a passive, stochastic process. Contrary this notion, we found that binding IgG-opsonized particles Fcγ receptors was inhibited in macrophages, dendritic and microglial cells agents interfere with actin assembly or disassembly. Changes the lateral mobility receptors—assessed single-particle tracking—or microelasticity membrane—determined atomic-force microscopy—could not account for effects disruption on particle binding. Instead,...
During mitosis in most eukaryotic cells, chromosomes align and form a metaphase plate halfway between the spindle poles, about which they exhibit oscillatory movement. These movements are accompanied by changes distance sister kinetochores, commonly referred to as breathing. We developed live cell imaging assay combined with computational image analysis quantify properties dynamics of kinetochores three dimensions. show that baseline oscillation breathing speeds late prometaphase set...
During T cell activation, biomolecular condensates form at the immunological synapse (IS) through multivalency-driven phase separation of LAT, Grb2, Sos1, SLP-76, Nck, and WASP. These move radially IS, traversing successive radially-oriented concentric actin networks. To understand this movement, we biochemically reconstituted LAT with actomyosin filaments. We found that basic regions Nck N-WASP/WASP promote association co-movement actin, indicating conversion weak individual affinities to...
Nuclear lamin isoforms form fibrous meshworks associated with nuclear pore complexes (NPCs). Using datasets prepared from subpixel and segmentation analyses of 3D–structured illumination microscopy images WT isoform knockout mouse embryo fibroblasts, we determined high precision the spatial association NPCs specific fibers. These relationships are retained in enlarged Lmna−/− Lmnb1−/− fibroblast nuclei. Cryo-ET observations reveal that filaments composing fibers contact nucleoplasmic ring...
Abstract The epidermal growth factor receptor (EGFR) is a central regulator of cell physiology. EGFR activated by ligand binding, triggering dimerization, activation kinase activity, and intracellular signaling. transiently confined within various plasma membrane nanodomains, yet how this may contribute to regulation binding poorly understood. To resolve nanoscale compartmentalization gates we developed single-particle tracking methods track the mobility ligand-bound total EGFR, in...
The actin cortex plays a large role in regulating the dynamic organization of cell surface receptors, which turn regulates their signaling. However, many receptors have short intracellular domains and no known link to cortical actin. In this work, we identified beta1-integrin subunit several tetraspanins, namely CD9, CD81 CD151, as part hitherto unknown molecular between receptor CD36 We found that vascular endothelial cells is recruited into complexes/nanodomains containing these proteins,...
One of the major challenges in multiple particle tracking is capture extremely heterogeneous movements objects crowded scenes. The presence numerous assignment candidates expected range motion makes ambiguous and induces false positives. Lowering ambiguity by reducing search range, on other hand, not an option, as this would increase rate negatives. We propose here a piecewise-stationary model (PMM) for transport along iterative smoother that exploits recursive rounds forward backward...
Nanoclustering is an emerging organizational principle for membrane-associated proteins. The functional consequences of nanoclustering receptor signaling remain largely unknown. Here, we applied quantitative multi-channel high- and super-resolution imaging to analyze the endothelial cell surface CD36, clustering which upon binding multivalent ligands, such as anti-angiogenic factor thrombospondin-1 (TSP-1), thought be crucial signaling. We found that a substantial fraction unligated CD36...
The dynamic nanoscale organization of cell surface receptors plays an important role in signaling. We determine this and its relation to activation VEGF receptor-2 (VEGFR-2), a critical receptor tyrosine kinase endothelial cells (ECs), by combining single-molecule imaging endogenous VEGFR-2 live ECs with multiscale computational analysis. find that can be mobile or exhibit restricted mobility monomeric non-monomeric, complex interplay between the two. This basal heterogeneity results...
ABSTRACT Keratin intermediate filaments (IFs) are the major cytoskeletal component in epithelial cells. The dynamics of keratin IFs have been described to depend mostly on actin cytoskeleton, but rapid transport fully polymerized has not reported. In this work, we used a combination photoconversion experiments and clustered regularly interspaced short palindromic repeats/clustered repeats–associated protein 9 genome editing study role microtubules microtubule motors filament transport. We...
Multivalent complexes of endothelial adhesion receptors (e.g., selectins) engage leukocytes to orchestrate their migration inflamed tissues. Proper anchorage and sufficient density (clustering) are required for efficient leukocyte capture rolling. We demonstrate that a polarized spectrin network dictates the stability cytoskeleton, which is attached apical membrane, at least in part, by abundant transmembrane protein CD44. Single-particle tracking revealed CD44 undergoes prolonged periods...