Filter
Associated Lab
- Aso Lab (1) Apply Aso Lab filter
- Remove Betzig Lab filter Betzig Lab
- Bock Lab (1) Apply Bock Lab filter
- Clapham Lab (1) Apply Clapham Lab filter
- Fetter Lab (2) Apply Fetter Lab filter
- Harris Lab (7) Apply Harris Lab filter
- Hess Lab (8) Apply Hess Lab filter
- Ji Lab (11) Apply Ji Lab filter
- Lavis Lab (8) Apply Lavis Lab filter
- Lippincott-Schwartz Lab (6) Apply Lippincott-Schwartz Lab filter
- Liu (Zhe) Lab (6) Apply Liu (Zhe) Lab filter
- Magee Lab (2) Apply Magee Lab filter
- Rubin Lab (1) Apply Rubin Lab filter
- Saalfeld Lab (2) Apply Saalfeld Lab filter
- Schreiter Lab (1) Apply Schreiter Lab filter
- Shroff Lab (9) Apply Shroff Lab filter
- Singer Lab (1) Apply Singer Lab filter
- Svoboda Lab (2) Apply Svoboda Lab filter
- Tjian Lab (4) Apply Tjian Lab filter
- Turner Lab (1) Apply Turner Lab filter
Associated Project Team
Publication Date
- 2024 (1) Apply 2024 filter
- 2023 (4) Apply 2023 filter
- 2022 (3) Apply 2022 filter
- 2021 (2) Apply 2021 filter
- 2020 (4) Apply 2020 filter
- 2019 (7) Apply 2019 filter
- 2018 (6) Apply 2018 filter
- 2017 (8) Apply 2017 filter
- 2016 (12) Apply 2016 filter
- 2015 (11) Apply 2015 filter
- 2014 (8) Apply 2014 filter
- 2013 (4) Apply 2013 filter
- 2012 (5) Apply 2012 filter
- 2011 (7) Apply 2011 filter
- 2010 (3) Apply 2010 filter
- 2009 (2) Apply 2009 filter
- 2008 (8) Apply 2008 filter
- 2007 (2) Apply 2007 filter
- 2006 (1) Apply 2006 filter
- 2005 (1) Apply 2005 filter
- 1995 (1) Apply 1995 filter
- 1994 (2) Apply 1994 filter
- 1993 (2) Apply 1993 filter
- 1992 (4) Apply 1992 filter
- 1991 (2) Apply 1991 filter
Type of Publication
110 Publications
Showing 21-30 of 110 resultsA microscope has a light source for generating a light beam having a wavelength, λ, and beam-forming optics configured for receiving the light beam and generating a Bessel-like beam that is directed into a sample. The beam-forming optics include an excitation objective having an axis oriented in a first direction. Imaging optics are configured for receiving light from a position within the sample that is illuminated by the Bessel-like beam and for imaging the received light on a detector. The imaging optics include a detection objective having an axis oriented in a second direction that is non-parallel to the first direction. A detector is configured for detecting signal light received by the imaging optics, and an aperture mask is positioned.
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.
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.
Commentary: Introduced the adiabatically tapered single mode fiber probe to near-field scanning optical microscopy which, together with shear force feedback, made the technique a practical reality. Although earlier claims of superresolution via near-field microscopy existed for nearly a decade, this paper was the first to convincingly break Abbe’s limit with visible light, as demonstrated by reproducibly resolving known, complex nanoscale patterns having features separated by much less than the wavelength. Whereas our fiber probe and shear force technologies were soon widely adopted and key to many novel applications (see above), the earlier methods proved to be technological dead ends, never achieving the results of their original claims. This experience taught me the most valuable lesson of my career: while it’s bad to bullshit others, it’s even worse to bullshit yourself. It’s a lesson sadly unheeded by many current practitioners of superresolution microscopy.
Fluorogenic molecules are important tools for advanced biochemical and biological experiments. The extant collection of fluorogenic probes is incomplete, however, leaving regions of the electromagnetic spectrum unutilized. Here, we synthesize green-excited fluorescent and fluorogenic analogues of the classic fluorescein and rhodamine 110 fluorophores by replacement of the xanthene oxygen with a quaternary carbon. These anthracenyl "carbofluorescein" and "carborhodamine 110" fluorophores exhibit excellent fluorescent properties and can be masked with enzyme- and photolabile groups to prepare high-contrast fluorogenic molecules useful for live cell imaging experiments and super-resolution microscopy. Our divergent approach to these red-shifted dye scaffolds will enable the preparation of numerous novel fluorogenic probes with high biological utility.
Neural circuit assembly features simultaneous targeting of numerous neuronal processes from constituent neuron types, yet the dynamics is poorly understood. Here, we use the Drosophila olfactory circuit to investigate dynamic cellular processes by which olfactory receptor neurons (ORNs) target axons precisely to specific glomeruli in the ipsi- and contralateral antennal lobes. Time-lapse imaging of individual axons from 30 ORN types revealed a rich diversity in extension speed, innervation timing, and ipsilateral branch locations and identified that ipsilateral targeting occurs via stabilization of transient interstitial branches. Fast imaging using adaptive optics-corrected lattice light-sheet microscopy showed that upon approaching target, many ORN types exhibiting "exploring branches" consisted of parallel microtubule-based terminal branches emanating from an F-actin-rich hub. Antennal nerve ablations uncovered essential roles for bilateral axons in contralateral target selection and for ORN axons to facilitate dendritic refinement of postsynaptic partner neurons. Altogether, these observations provide cellular bases for wiring specificity establishment.
The signal and resolution during in vivo imaging of the mouse brain is limited by sample-induced optical aberrations. We find that, although the optical aberrations can vary across the sample and increase in magnitude with depth, they remain stable for hours. As a result, two-photon adaptive optics can recover diffraction-limited performance to depths of 450 μm and improve imaging quality over fields of view of hundreds of microns. Adaptive optical correction yielded fivefold signal enhancement for small neuronal structures and a threefold increase in axial resolution. The corrections allowed us to detect smaller neuronal structures at greater contrast and also improve the signal-to-noise ratio during functional Ca(2+) imaging in single neurons.
Lattice light sheet microscopy excels at the non-invasive imaging of three-dimensional (3D) dynamic processes at high spatiotemporal resolution within cells and developing embryos. Recently, several papers have called into question the performance of lattice light sheets relative to the Gaussian sheets most common in light sheet microscopy. Here we undertake a comprehensive theoretical and experimental analysis of various forms of light sheet microscopy which both demonstrates and explains why lattice light sheets provide significant improvements in resolution and photobleaching reduction. The analysis provides a procedure to select the correct light sheet for a desired experiment and specifies the processing that maximizes the use of all fluorescence generated within the light sheet excitation envelope for optimal resolution while minimizing image artifacts and photodamage. Development of a new type of “harmonic balanced” lattice light sheet is shown to improve performance at all spatial frequencies within its 3D resolution limits and maintains this performance over lengthened propagation distances allowing for expanded fields of view.
A distance regulation method has been developed to enhance the reliability, versatility, and ease of use of near-field scanning optical microscopy (NSOM). The method relies on the detection of shear forces between the end of a near-field probe and the sample of interest. The system can be used solely for distance regulation in NSOM, for simultaneous shear force and near-field imaging, or for shear force microscopy alone. In the latter case, uncoated optical fiber probes are found to yield images with consistently high resolution.
Commentary: To exploit the evanescent field that is the source of high resolution in near-field microscopy, the probe must be exceptionally close to the sample: 10 nm away for 30-50 nm resolution. Here we introduced a distance regulation mechanism based on transverse shear forces between the end of a dithered near-field probe and the sample, which permitted even samples of modest topography to be imaged. Simple, reliable, noninvasive, and applicable to a wide range of samples from whole fixed cells to semiconductor devices, shear force microscopy was a key enabling technology for near-field optics, and soon widely implemented.
Mechano-transduction is an emerging but still poorly understood component of T cell activation. Here we investigated the ligand-dependent contribution made by contractile actomyosin arcs populating the peripheral supramolecular activation cluster (pSMAC) region of the immunological synapse (IS) to T cell receptor (TCR) microcluster transport and proximal signaling in primary mouse T cells. Using super resolution microscopy, OT1-CD8+ mouse T cells, and two ovalbumin (OVA) peptides with different affinities for the TCR, we show that the generation of organized actomyosin arcs depends on ligand potency and the ability of myosin 2 to contract actin filaments. While weak ligands induce disorganized actomyosin arcs, strong ligands result in organized actomyosin arcs that correlate well with tension-sensitive CasL phosphorylation and the accumulation of ligands at the IS center. Blocking myosin 2 contractility greatly reduces the difference in the extent of Src and LAT phosphorylation observed between the strong and the weak ligand, arguing that myosin 2-dependent force generation within actin arcs contributes to ligand discrimination. Together, our data are consistent with the idea that actomyosin arcs in the pSMAC region of the IS promote a mechano-chemical feedback mechanism that amplifies the accumulation of critical signaling molecules at the IS.
Within cells, the spatial compartmentalization of thousands of distinct proteins serves a multitude of diverse biochemical needs. Correlative super-resolution (SR) fluorescence and electron microscopy (EM) can elucidate protein spatial relationships to global ultrastructure, but has suffered from tradeoffs of structure preservation, fluorescence retention, resolution, and field of view. We developed a platform for three-dimensional cryogenic SR and focused ion beam-milled block-face EM across entire vitreously frozen cells. The approach preserves ultrastructure while enabling independent SR and EM workflow optimization. We discovered unexpected protein-ultrastructure relationships in mammalian cells including intranuclear vesicles containing endoplasmic reticulum-associated proteins, web-like adhesions between cultured neurons, and chromatin domains subclassified on the basis of transcriptional activity. Our findings illustrate the value of a comprehensive multimodal view of ultrastructural variability across whole cells.