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
From single molecules to life: microscopy at the nanoscale.
TL;DR: A theoretical overview of the techniques and underlying physics are presented, followed by a practical guide to all of the facets involved in designing a super-resolution experiment, including an approachable explanation of the photochemistry involved, labeling methods available, and sample preparation procedures.
Journal ArticleDOI
MINFLUX nanometer-scale 3D imaging and microsecond-range tracking on a common fluorescence microscope.
Roman Schmidt,Tobias Weihs,Christian A. Wurm,Isabelle Jansen,Jasmin Rehman,Steffen J. Sahl,Stefan W. Hell +6 more
TL;DR: In this paper, the authors show that MINFLUX with a standard microscope stand can achieve 3D resolution of 2.2 and 2.4 degrees of freedom (DOF) in the focal plane and 1.9 degrees along the optic axis.
Journal ArticleDOI
Stochastic optical reconstruction microscopy (STORM) in comparison with stimulated emission depletion (STED) and other imaging methods
Johnny Tam,David Merino +1 more
TL;DR: The choice between STORM and STED will depend not only on the specific application, but also on the users' ability to understand and optimize the various parameters ranging from sample preparation to image acquisition, which determine the quality of the final image.
Journal ArticleDOI
Reporting from the Field: Genetically Encoded Fluorescent Reporters Uncover Signaling Dynamics in Living Biological Systems
Sohum Mehta,Jin Zhang +1 more
TL;DR: This review of genetically encoded fluorescent reporters discusses some of the unique insights that they continue to provide regarding the spatial organization and dynamic regulation of intracellular signaling networks, and explores the more recent push to expand the scope of biological phenomena that can be monitored using these reporters.
Journal ArticleDOI
Fast high-resolution 3D total internal reflection fluorescence microscopy by incidence angle scanning and azimuthal averaging
Jérôme Boulanger,Charles Gueudry,Daniel Münch,Bertrand Cinquin,Perrine Paul-Gilloteaux,Sabine Bardin,Christophe Guérin,Fabrice Senger,Laurent Blanchoin,Jean Salamero +9 more
TL;DR: A method to recover 3D volumes from images obtained using several total internal reflection fluorescence (TIRF) incidence angles at dense regime of acquisition and achieves unprecedented 50-nm axial resolution over a range of 800 nm above the coverslip.
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