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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Mcauliffe, Matthew Fu, Yan Winter, Peter W. Rojas, Raul Wang, Victor Patterson, George H. |
| Description | Author Affiliation: Fu Y ( Section on Biophotonics, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD 20892); Winter PW ( Section on High-Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD 20892); Rojas R ( Section on Biological Chemistry, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892); Wang V ( Biomedical Imaging Research Services Section, Division of Computational Bioscience, Center for Information Technology, National Institutes of Health, Bethesda, MD 20892.); McAuliffe M ( Biomedical Imaging Research Services Section, Division of Computational Bioscience, Center for Information Technology, National Institutes of Health, Bethesda, MD 20892.); Patterson GH ( Section on Biophotonics, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD 20892); |
| Abstract | We report superresolution optical sectioning using a multiangle total internal reflection fluorescence (TIRF) microscope. TIRF images were constructed from several layers within a normal TIRF excitation zone by sequentially imaging and photobleaching the fluorescent molecules. The depth of the evanescent wave at different layers was altered by tuning the excitation light incident angle. The angle was tuned from the highest (the smallest TIRF depth) toward the critical angle (the largest TIRF depth) to preferentially photobleach fluorescence from the lower layers and allow straightforward observation of deeper structures without masking by the brighter signals closer to the coverglass. Reconstruction of the TIRF images enabled 3D imaging of biological samples with 20-nm axial resolution. Two-color imaging of epidermal growth factor (EGF) ligand and clathrin revealed the dynamics of EGF-activated clathrin-mediated endocytosis during internalization. Furthermore, Bayesian analysis of images collected during the photobleaching step of each plane enabled lateral superresolution (<100 nm) within each of the sections. |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 16 |
| Volume Number | 113 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 2016-04-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Clathrin Metabolism Endocytosis Physiology Epidermal Growth Factor Fluorescent Dyes Chemistry Imaging, Three-Dimensional Photobleaching Cell Line Microscopy, Fluorescence Research Support, N.I.H., Intramural Multidisciplinary |
| Content Type | Text |
| Resource Type | Article |
| Subject | Multidisciplinary |
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