Imaging intracellular fluorescent proteins at nanometer resolution.
Eric Betzig,George H. Patterson,Rachid Sougrat,O. Wolf Lindwasser,Scott G. Olenych,Juan S. Bonifacino,Michael W. Davidson,Jennifer Lippincott-Schwartz,Harald F. Hess +8 more
Reads0
Chats0
TLDR
This work introduced a method for optically imaging intracellular proteins at nanometer spatial resolution and used this method to image specific target proteins in thin sections of lysosomes and mitochondria and in fixed whole cells to image retroviral protein Gag at the plasma membrane.Abstract:
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.read more
Citations
More filters
Journal ArticleDOI
Super-resolution fluorescence imaging of organelles in live cells with photoswitchable membrane probes.
Sang Hee Shim,Chenglong Xia,Guisheng Zhong,Guisheng Zhong,Hazen P. Babcock,Joshua C. Vaughan,Joshua C. Vaughan,Bo Huang,Xun Wang,Cheng Xu,Guo-Qiang Bi,Xiaowei Zhuang +11 more
TL;DR: This work identified photoswitchable membrane probes and obtained super-resolution fluorescence images of cellular membranes, and demonstrated the photoswitching capabilities of eight commonly used membrane probes, each specific to the plasma membrane, mitochondria, the endoplasmic recticulum (ER) or lysosomes.
Journal ArticleDOI
Photochromism into nanosystems: towards lighting up the future nanoworld.
Ling Wang,Quan Li +1 more
TL;DR: This Review provides an account of the recent advancements in reversible photocontrol of the structures and functions of photochromic nanosystems and their applications and outlines the challenges that need to be addressed and the opportunities that can be tapped into.
Journal ArticleDOI
Compact, light-weight and cost-effective microscope based on lensless incoherent holography for telemedicine applications
Onur Mudanyali,Derek Tseng,Chulwoo Oh,Serhan O. Isikman,Ikbal Sencan,Waheb Bishara,Cetin Oztoprak,Sungkyu Seo,Bahar Khademhosseini,Aydogan Ozcan +9 more
TL;DR: This lensless incoherent holographic microscope has orders-of-magnitude improved light collection efficiency and is very robust to mechanical misalignments it may offer a cost-effective tool especially for telemedicine applications involving various global health problems in resource limited settings.
Journal ArticleDOI
Stimulated emission depletion (STED) nanoscopy of a fluorescent protein-labeled organelle inside a living cell
TL;DR: Imaging individual structural elements of the ER revealed a focal plane resolution of <50 nm inside the living cell, corresponding to a 4-fold improvement over that of a confocal microscope and a 16-fold reduction in the focal-spot cross-sectional area.
Journal ArticleDOI
Lensfree on-chip microscopy over a wide field-of-view using pixel super-resolution
TL;DR: A sub-pixel shifting based super-resolution algorithm is implemented to effectively recover much higher resolution digital holograms of the objects, permitting sub-micron spatial resolution to be achieved across the entire sensor chip active area.
References
More filters
疟原虫var基因转换速率变化导致抗原变异[英]/Paul H, Robert P, Christodoulou Z, et al//Proc Natl Acad Sci U S A
TL;DR: PfPMP1)与感染红细胞、树突状组胞以及胎盘的单个或多个受体作用,在黏附及免疫逃避中起关键的作�ly.
Journal ArticleDOI
Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy.
TL;DR: Lateral resolution that exceeds the classical diffraction limit by a factor of two is achieved by using spatially structured illumination in a wide‐field fluorescence microscope with strikingly increased clarity compared to both conventional and confocal microscopes.
Journal ArticleDOI
Precise nanometer localization analysis for individual fluorescent probes
TL;DR: A localization algorithm motivated from least-squares fitting theory is constructed and tested both on image stacks of 30-nm fluorescent beads and on computer-generated images (Monte Carlo simulations), and results show good agreement with the derived precision equation.
Journal ArticleDOI
Nonlinear structured-illumination microscopy: Wide-field fluorescence imaging with theoretically unlimited resolution
TL;DR: Experimental results show that a 2D point resolution of <50 nm is possible on sufficiently bright and photostable samples, and a recently proposed method in which the nonlinearity arises from saturation of the excited state is experimentally demonstrated.
Journal ArticleDOI
Myosin V Walks Hand-Over-Hand: Single Fluorophore Imaging with 1.5-nm Localization
Ahmet Yildiz,Joseph N. Forkey,Sean A. McKinney,Taekjip Ha,Taekjip Ha,Yale E. Goldman,Paul R. Selvin,Paul R. Selvin +7 more
TL;DR: The results strongly support a hand-over-hand model of motility, not an inchworm model, which moves processively on actin.
Related Papers (5)
Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM).
Ultra-High Resolution Imaging by Fluorescence Photoactivation Localization Microscopy
Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy
Stefan W. Hell,Jan Wichmann +1 more