A5002646367- Advanced Fluorescence Microscopy Techniques
- Advanced Electron Microscopy Techniques and Applications
- Near-Field Optical Microscopy
- Cell Image Analysis Techniques
- Lipid Membrane Structure and Behavior
- Genetics, Aging, and Longevity in Model Organisms
- Photosynthetic Processes and Mechanisms
- Cellular transport and secretion
- Force Microscopy Techniques and Applications
- Pancreatic function and diabetes
- Microtubule and mitosis dynamics
- Photoacoustic and Ultrasonic Imaging
- Advanced X-ray Imaging Techniques
- CAR-T cell therapy research
- Sperm and Testicular Function
- Biotin and Related Studies
- Optical Coherence Tomography Applications
- Erythrocyte Function and Pathophysiology
- Bacteriophages and microbial interactions
- Virus-based gene therapy research
- Digital Holography and Microscopy
- Image Processing Techniques and Applications
- Spectroscopy Techniques in Biomedical and Chemical Research
- Heat shock proteins research
- Reproductive Biology and Fertility
Yale University
2016-2025
Westlake University
2021-2024
University of New Haven
2018-2021
Chinese Academy of Sciences
2012-2019
Institute of Biophysics
2012-2019
Huazhong University of Science and Technology
2010-2013
Fluorescence nanoscopy, or super-resolution microscopy, has become an important tool in cell biological research. However, because of its usually inferior resolution the depth direction (50–80 nm) and rapidly deteriorating thick samples, practical application been effectively limited to two dimensions thin samples. Here, we present development whole-cell 4Pi single-molecule switching nanoscopy (W-4PiSMSN), optical nanoscope that allows imaging three-dimensional (3D) structures at 10- 20-nm...
All living organisms deploy cell-autonomous defenses to combat infection. In plants and animals, large supramolecular complexes often activate immune proteins for protection. this work, we resolved the native structure of a massive host-defense complex that polymerizes 30,000 guanylate-binding (GBPs) over surface gram-negative bacteria inside human cells. Construction giant nanomachine took several minutes remained stable hours, required guanosine triphosphate hydrolysis, recruited four GBPs...
The endoplasmic reticulum (ER) is composed of interconnected membrane sheets and tubules. Superresolution microscopy recently revealed densely packed, rapidly moving ER tubules mistaken for by conventional light microscopy, highlighting the importance revisiting classical views structure with high spatiotemporal resolution in living cells. In this study, we use live-cell stimulated emission depletion (STED) to survey architecture at 50-nm resolution. We determine nanoscale dimensions first...
Reversibly switchable fluorescent proteins (RSFPs) have attracted widespread interest for emerging techniques including repeated tracking of protein behavior and superresolution microscopy. Among the limited number RSFPs available, only Dronpa is widely employed most cell biology applications due to its monomeric other favorable photochemical properties. Here we developed a series green with beneficial optical characteristics such as high photon output per switch, photostability, broad range...
We demonstrate the use of cryogenic super-resolution correlative light and electron microscopy (csCLEM) to precisely determine spatial relationship between proteins their native cellular structures. Several fluorescent (FPs) were found be photoswitchable emitted far more photons under our imaging condition, resulting in higher localization precision which is comparable ambient imaging. Vitrified specimens prepared by high pressure freezing cryo-sectioning maintain a near-native state with...
Arabidopsis hypersensitive-induced reaction (AtHIR) proteins function in plant innate immunity. However, the underlying mechanisms by which AtHIRs participate immunity remain elusive. Here, using VA-TIRFM and FLIM-FRET, we revealed that AtHIR1 is present membrane microdomains co-localizes with microdomain marker REM1.3. Single-particle tracking analysis cytoskeleton, especially microtubules, restrict lateral mobility of at plasma facilitate its oligomerization. Furthermore, protein proximity...
Significance Type III protein secretion systems are essential virulence factors for many bacterial pathogens. Cryo-electron microscopy has provided important details about the architecture and molecular organization of type machine in isolation or fixed samples. However, little information is available its individual substructures live bacteria. Using 2D 3D single-molecule switching superresolution microscopy, we have visualized machines bacteria obtained unique insight into assembly...
Single-molecule switching nanoscopy overcomes the diffraction limit of light by stochastically single fluorescent molecules on and off, then localizing their positions individually. Recent advances in this technique have greatly accelerated data acquisition speed improved temporal resolution super-resolution imaging. However, it has not been quantified whether increase comes at cost compromised image quality. The spatial depends many factors, among which laser intensity camera are two most...
The majority of gels exhibit nanoscale spatial variations in crosslink density. We present the first 3D super-resolution microscopy images dye tagged cross-link distributions microgels and hydrogels. morphology features never imaged previously microgels, are revealed.
Three-dimensional structured illumination microscopy (3D-SIM) provides excellent optical sectioning and doubles the resolution in all dimensions compared with wide-field microscopy. However, its much lower axial results blurred fine details that direction overall image distortion. Here we present 4Pi-SIM, a substantial revamp of I5S synergizes 3D-SIM interferometric to achieve isotropic through interference both detection wavefronts. We evaluate performance 4Pi-SIM by imaging various...
Golgin proteins have long been suspected to be organizers of the Golgi stack. Using three-dimensional super-resolution microscopy, we comprehensively localize human golgin family at rim apparatus 10-20 nm resolution in situ. Unexpectedly, find that golgins are precisely organized into a tetraplex with four discrete layers, each containing specific set golgins. We observe no inside stack between its membrane-bound cisternae. Biochemically characterizing most as isolated proteins, they form...