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  • 1
    Publication Date: 1992-07-10
    Description: The near-field optical interaction between a sharp probe and a sample of interest can be exploited to image, spectroscopically probe, or modify surfaces at a resolution (down to approximately 12 nm) inaccessible by traditional far-field techniques. Many of the attractive features of conventional optics are retained, including noninvasiveness, reliability, and low cost. In addition, most optical contrast mechanisms can be extended to the near-field regime, resulting in a technique of considerable versatility. This versatility is demonstrated by several examples, such as the imaging of nanometric-scale features in mammalian tissue sections and the creation of ultrasmall, magneto-optic domains having implications for highdensity data storage. Although the technique may find uses in many diverse fields, two of the most exciting possibilities are localized optical spectroscopy of semiconductors and the fluorescence imaging of living cells.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Betzig, E -- Trautman, J K -- New York, N.Y. -- Science. 1992 Jul 10;257(5067):189-95.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/17794749" target="_blank"〉PubMed〈/a〉
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 2
    Publication Date: 1991-03-22
    Description: In near-field scanning optical microscopy, a light source or detector with dimensions less than the wavelength (lambda) is placed in close proximity (lambda/50) to a sample to generate images with resolution better than the diffraction limit. A near-field probe has been developed that yields a resolution of approximately 12 nm ( approximately lambda/43) and signals approximately 10(4)- to 10(6)-fold larger than those reported previously. In addition, image contrast is demonstrated to be highly polarization dependent. With these probes, near-field microscopy appears poised to fulfill its promise by combining the power of optical characterization methods with nanometric spatial resolution.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Betzig, E -- Trautman, J K -- Harris, T D -- Weiner, J S -- Kostelak, R L -- New York, N.Y. -- Science. 1991 Mar 22;251(5000):1468-70.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/17779440" target="_blank"〉PubMed〈/a〉
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    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 3
    Publication Date: 2006-08-12
    Description: We introduce a method for optically imaging intracellular proteins at nanometer spatial resolution. Numerous sparse subsets of photoactivatable fluorescent protein molecules were activated, localized (to approximately 2 to 25 nanometers), and then bleached. The aggregate position information from all subsets was then assembled into a superresolution image. We used this method--termed photoactivated localization microscopy--to image specific target proteins in thin sections of lysosomes and mitochondria; in fixed whole cells, we imaged vinculin at focal adhesions, actin within a lamellipodium, and the distribution of the retroviral protein Gag at the plasma membrane.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Betzig, Eric -- Patterson, George H -- Sougrat, Rachid -- Lindwasser, O Wolf -- Olenych, Scott -- Bonifacino, Juan S -- Davidson, Michael W -- Lippincott-Schwartz, Jennifer -- Hess, Harald F -- New York, N.Y. -- Science. 2006 Sep 15;313(5793):1642-5. Epub 2006 Aug 10.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, VA 20147, USA. betzige@janelia.hhmi.org〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/16902090" target="_blank"〉PubMed〈/a〉
    Keywords: Actins/analysis ; Algorithms ; Animals ; COS Cells ; Cell Line ; Cell Membrane/*chemistry ; Cercopithecus aethiops ; Fluorescence ; Focal Adhesions/chemistry ; Gene Products, gag/analysis ; Hiv-1 ; Light ; Luminescent Proteins/*analysis ; Lysosomes/chemistry ; Microscopy/*methods ; Mitochondria/chemistry ; *Nanotechnology ; Organelles/*chemistry ; Photobleaching ; Proteins/*analysis ; Pseudopodia/chemistry ; Recombinant Fusion Proteins/*analysis ; Vinculin/analysis
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    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 4
    Publication Date: 1995-10-27
    Description: Near-field scanning optical microscopy of phospholipid monolayers doped with fluorescent lipid analogs reveals previously undescribed features in various phases, including a concentration gradient at the liquid-expanded/liquid-condensed domain boundary and weblike structures in the solid-condensed phase. Presumably, the web structures are grain boundaries between crystalline solid lipid. These structures are strongly modulated by the addition of low concentrations of cholesterol and ganglioside GM1 in the monolayer.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Hwang, J -- Tamm, L K -- Bohm -- Ramalingam, T S -- Betzig, E -- Edidin, M -- AI14584/AI/NIAID NIH HHS/ -- AI30557/AI/NIAID NIH HHS/ -- DK44375/DK/NIDDK NIH HHS/ -- R37 AI030557/AI/NIAID NIH HHS/ -- New York, N.Y. -- Science. 1995 Oct 27;270(5236):610-4.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Department of Biology, Johns Hopkins University, Baltimore, MD 21218, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/7570018" target="_blank"〉PubMed〈/a〉
    Keywords: 1,2-Dipalmitoylphosphatidylcholine/*chemistry ; Boron Compounds ; Cholesterol/*chemistry ; Fluorescent Dyes ; G(M1) Ganglioside/*chemistry ; Microscopy/*methods ; Microscopy, Fluorescence ; Phosphatidylcholines ; Phospholipids/*chemistry
    Print ISSN: 0036-8075
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  • 5
    Publication Date: 1993-11-26
    Description: Individual carbocyanine dye molecules in a sub-monolayer spread have been imaged with near-field scanning optical microscopy. Molecules can be repeatedly detected and spatially localized (to approximately lambda/50 where lambda is the wavelength of light) with a sensitivity of at least 0.005 molecules/(Hz)(1/2) and the orientation of each molecular dipole can be determined. This information is exploited to map the electric field distribution in the near-field aperture with molecular spatial resolution.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Betzig, E -- Chichester, R J -- New York, N.Y. -- Science. 1993 Nov 26;262(5138):1422-5.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/17736823" target="_blank"〉PubMed〈/a〉
    Print ISSN: 0036-8075
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    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 6
    Publication Date: 1994-06-17
    Description: Luminescent centers with sharp (〈0.07 millielectron volt), spectrally distinct emission lines were imaged in a GaAs/AIGaAs quantum well by means of low-temperature near-field scanning optical microscopy. Temperature, magnetic field, and linewidth measurements establish that these centers arise from excitons laterally localized at interface fluctuations. For sufficiently narrow wells, virtually all emission originates from such centers. Near-field microscopy/spectroscopy provides a means to access energies and homogeneous line widths for the individual eigenstates of these centers, and thus opens a rich area of physics involving quantum resolved systems.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Hess, H F -- Betzig, E -- Harris, T D -- Pfeiffer, L N -- West, K W -- New York, N.Y. -- Science. 1994 Jun 17;264(5166):1740-5.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/17839907" target="_blank"〉PubMed〈/a〉
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  • 7
    Publication Date: 2012-02-11
    Description: Apical constriction changes cell shapes, driving critical morphogenetic events, including gastrulation in diverse organisms and neural tube closure in vertebrates. Apical constriction is thought to be triggered by contraction of apical actomyosin networks. We found that apical actomyosin contractions began before cell shape changes in both Caenorhabitis elegans and Drosophila. In C. elegans, actomyosin networks were initially dynamic, contracting and generating cortical tension without substantial shrinking of apical surfaces. Apical cell-cell contact zones and actomyosin only later moved increasingly in concert, with no detectable change in actomyosin dynamics or cortical tension. Thus, apical constriction appears to be triggered not by a change in cortical tension, but by dynamic linking of apical cell-cell contact zones to an already contractile apical cortex.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3298882/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉   〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3298882/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Roh-Johnson, Minna -- Shemer, Gidi -- Higgins, Christopher D -- McClellan, Joseph H -- Werts, Adam D -- Tulu, U Serdar -- Gao, Liang -- Betzig, Eric -- Kiehart, Daniel P -- Goldstein, Bob -- R01 GM033830/GM/NIGMS NIH HHS/ -- R01 GM083071/GM/NIGMS NIH HHS/ -- R01 GM083071-01A1/GM/NIGMS NIH HHS/ -- R01 GM083071-02/GM/NIGMS NIH HHS/ -- R01 GM083071-02S1/GM/NIGMS NIH HHS/ -- R01 GM083071-03/GM/NIGMS NIH HHS/ -- R01 GM083071-04/GM/NIGMS NIH HHS/ -- R01 GM33830/GM/NIGMS NIH HHS/ -- Howard Hughes Medical Institute/ -- New York, N.Y. -- Science. 2012 Mar 9;335(6073):1232-5. doi: 10.1126/science.1217869. Epub 2012 Feb 9.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/22323741" target="_blank"〉PubMed〈/a〉
    Keywords: Actomyosin/chemistry/*physiology ; Animals ; Caenorhabditis elegans/*cytology/*embryology ; Cell Membrane/physiology/ultrastructure ; *Cell Shape ; Computer Simulation ; Cytoskeleton/physiology/ultrastructure ; Drosophila melanogaster/*cytology/*embryology ; Embryo, Nonmammalian/cytology/physiology ; Fluorescence Recovery After Photobleaching ; *Gastrulation ; Intercellular Junctions/physiology/ultrastructure ; Mechanical Phenomena ; Models, Biological ; Morphogenesis ; Myosins/chemistry/physiology
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  • 8
    Publication Date: 2013-03-23
    Description: Developmental signals such as Wnts are often presented to cells in an oriented manner. To examine the consequences of local Wnt signaling, we immobilized Wnt proteins on beads and introduced them to embryonic stem cells in culture. At the single-cell level, the Wnt-bead induced asymmetric distribution of Wnt-beta-catenin signaling components, oriented the plane of mitotic division, and directed asymmetric inheritance of centrosomes. Before cytokinesis was completed, the Wnt-proximal daughter cell expressed high levels of nuclear beta-catenin and pluripotency genes, whereas the distal daughter cell acquired hallmarks of differentiation. We suggest that a spatially restricted Wnt signal induces an oriented cell division that generates distinct cell fates at predictable positions relative to the Wnt source.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3966430/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉   〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3966430/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Habib, Shukry J -- Chen, Bi-Chang -- Tsai, Feng-Chiao -- Anastassiadis, Konstantinos -- Meyer, Tobias -- Betzig, Eric -- Nusse, Roel -- 102513/Wellcome Trust/United Kingdom -- GM063702/GM/NIGMS NIH HHS/ -- NS069375/NS/NINDS NIH HHS/ -- R01 GM030179/GM/NIGMS NIH HHS/ -- R01 GM063702/GM/NIGMS NIH HHS/ -- Howard Hughes Medical Institute/ -- New York, N.Y. -- Science. 2013 Mar 22;339(6126):1445-8. doi: 10.1126/science.1231077.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Department of Developmental Biology, Howard Hughes Medical Institute, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University, 265 Campus Drive, Stanford, CA 94305, USA. shabib@stanford.edu〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/23520113" target="_blank"〉PubMed〈/a〉
    Keywords: Animals ; *Asymmetric Cell Division ; *Cell Differentiation ; Cells, Cultured ; Centrosome/physiology ; Cytokinesis ; Embryonic Stem Cells/*cytology/*metabolism ; Gene Expression ; Homeodomain Proteins/genetics/metabolism ; Mice ; Mitosis ; Octamer Transcription Factor-3/genetics/metabolism ; Pluripotent Stem Cells/physiology ; Recombinant Proteins/metabolism ; Single-Cell Analysis ; Transcription Factors/genetics/metabolism ; Wnt Proteins/metabolism ; *Wnt Signaling Pathway ; Wnt3A Protein/*metabolism ; beta Catenin/metabolism
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    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 9
    Publication Date: 2014-10-25
    Description: Although fluorescence microscopy provides a crucial window into the physiology of living specimens, many biological processes are too fragile, are too small, or occur too rapidly to see clearly with existing tools. We crafted ultrathin light sheets from two-dimensional optical lattices that allowed us to image three-dimensional (3D) dynamics for hundreds of volumes, often at subsecond intervals, at the diffraction limit and beyond. We applied this to systems spanning four orders of magnitude in space and time, including the diffusion of single transcription factor molecules in stem cell spheroids, the dynamic instability of mitotic microtubules, the immunological synapse, neutrophil motility in a 3D matrix, and embryogenesis in Caenorhabditis elegans and Drosophila melanogaster. The results provide a visceral reminder of the beauty and the complexity of living systems.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4336192/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉   〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4336192/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Chen, Bi-Chang -- Legant, Wesley R -- Wang, Kai -- Shao, Lin -- Milkie, Daniel E -- Davidson, Michael W -- Janetopoulos, Chris -- Wu, Xufeng S -- Hammer, John A 3rd -- Liu, Zhe -- English, Brian P -- Mimori-Kiyosue, Yuko -- Romero, Daniel P -- Ritter, Alex T -- Lippincott-Schwartz, Jennifer -- Fritz-Laylin, Lillian -- Mullins, R Dyche -- Mitchell, Diana M -- Bembenek, Joshua N -- Reymann, Anne-Cecile -- Bohme, Ralph -- Grill, Stephan W -- Wang, Jennifer T -- Seydoux, Geraldine -- Tulu, U Serdar -- Kiehart, Daniel P -- Betzig, Eric -- GM33830/GM/NIGMS NIH HHS/ -- R01 GM033830/GM/NIGMS NIH HHS/ -- R01GM080370/GM/NIGMS NIH HHS/ -- R01HD37047/HD/NICHD NIH HHS/ -- RM01-GM61010/GM/NIGMS NIH HHS/ -- T32 GM007445/GM/NIGMS NIH HHS/ -- Howard Hughes Medical Institute/ -- Intramural NIH HHS/ -- New York, N.Y. -- Science. 2014 Oct 24;346(6208):1257998. doi: 10.1126/science.1257998. Epub 2014 Oct 23.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA. ; Coleman Technologies, Incorporated, Newtown Square, PA 19073, USA. ; National High Magnetic Field Laboratory and Department of Biological Science, Florida State University, Tallahassee, FL 32310, USA. ; Department of Biological Sciences, University of the Sciences, Philadelphia, PA 19104, USA. ; Laboratory of Cell Biology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA. ; Optical Image Analysis Unit, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan. ; Department of Pharmacology, University of Minnesota, Minneapolis, MN 55455, USA. ; Cell Biology and Metabolism Program, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA. Cambridge Institute for Medical Research, Addenbrooke's Hospital, Cambridge CB2 0XY, England, UK. ; Cell Biology and Metabolism Program, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA. ; Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA. ; Department of Biochemistry and Cellular and Molecular Biology, University of Tennessee, Knoxville, TN 37996, USA. ; Max Planck Institute for Molecular Cell Biology and Genetics, 01307 Dresden, Germany. Max Planck Institute for the Physics of Complex Systems, 01307 Dresden, Germany. ; Department of Molecular Biology and Genetics, Howard Hughes Medical Institute, Center for Cell Dynamics, Johns Hopkins School of Medicine, Baltimore, MD 21205, USA. ; Department of Biology, Duke University, Durham, NC 27708, USA. ; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA. betzige@janelia.hhmi.org.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/25342811" target="_blank"〉PubMed〈/a〉
    Keywords: Animals ; Caenorhabditis elegans/*embryology ; Cell Communication ; Drosophila melanogaster/*embryology ; Embryo, Nonmammalian/*ultrastructure ; Embryonic Stem Cells/ultrastructure ; Imaging, Three-Dimensional/*methods ; Mice ; Microscopy/*methods ; Molecular Imaging/*methods ; Spheroids, Cellular/ultrastructure
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  • 10
    Publication Date: 2015-09-01
    Description: Super-resolution fluorescence microscopy is distinct among nanoscale imaging tools in its ability to image protein dynamics in living cells. Structured illumination microscopy (SIM) stands out in this regard because of its high speed and low illumination intensities, but typically offers only a twofold resolution gain. We extended the resolution of live-cell SIM through two approaches: ultrahigh numerical aperture SIM at 84-nanometer lateral resolution for more than 100 multicolor frames, and nonlinear SIM with patterned activation at 45- to 62-nanometer resolution for approximately 20 to 40 frames. We applied these approaches to image dynamics near the plasma membrane of spatially resolved assemblies of clathrin and caveolin, Rab5a in early endosomes, and alpha-actinin, often in relationship to cortical actin. In addition, we examined mitochondria, actin, and the Golgi apparatus dynamics in three dimensions.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4659358/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉   〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4659358/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Li, Dong -- Shao, Lin -- Chen, Bi-Chang -- Zhang, Xi -- Zhang, Mingshu -- Moses, Brian -- Milkie, Daniel E -- Beach, Jordan R -- Hammer, John A 3rd -- Pasham, Mithun -- Kirchhausen, Tomas -- Baird, Michelle A -- Davidson, Michael W -- Xu, Pingyong -- Betzig, Eric -- GM-075252/GM/NIGMS NIH HHS/ -- R01 GM075252/GM/NIGMS NIH HHS/ -- Howard Hughes Medical Institute/ -- New York, N.Y. -- Science. 2015 Aug 28;349(6251):aab3500. doi: 10.1126/science.aab3500.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA. ; Key Laboratory of RNA Biology and Beijing Key Laboratory of Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China. College of Life Sciences, Central China Normal University, Wuhan 430079, Hubei, China. ; Key Laboratory of RNA Biology and Beijing Key Laboratory of Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China. ; Coleman Technologies, 5131 West Chester Pike, Newtown Square, PA 19073, USA. ; Cell Biology and Physiology Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA. ; Department of Cell Biology and Pediatrics, Harvard Medical School and Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115, USA. ; Cell Biology and Physiology Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA. National High Magnetic Field Laboratory and Department of Biological Science, Florida State University, Tallahassee, FL 32310, USA. ; National High Magnetic Field Laboratory and Department of Biological Science, Florida State University, Tallahassee, FL 32310, USA. ; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA. betzige@janelia.hhmi.org.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/26315442" target="_blank"〉PubMed〈/a〉
    Keywords: Actinin/analysis ; Actins/analysis ; Animals ; Cell Line ; Clathrin/analysis ; Clathrin-Coated Vesicles/chemistry/ultrastructure ; Coated Pits, Cell-Membrane/chemistry/ultrastructure ; Cytoskeleton/chemistry/metabolism/*ultrastructure ; *Endocytosis ; Endosomes/chemistry/ultrastructure ; Golgi Apparatus/ultrastructure ; Image Processing, Computer-Assisted ; Imaging, Three-Dimensional/instrumentation/*methods ; Microscopy, Fluorescence/instrumentation/*methods ; Mitochondria/chemistry/ultrastructure ; Organelles/chemistry/metabolism/*ultrastructure ; rab5 GTP-Binding Proteins/analysis
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    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
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