Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM).
TLDR
A high-resolution fluorescence microscopy method based on high-accuracy localization of photoswitchable fluorophores that can, in principle, reach molecular-scale resolution is developed.Abstract:
We have developed a high-resolution fluorescence microscopy method based on high-accuracy localization of photoswitchable fluorophores. In each imaging cycle, only a fraction of the fluorophores were turned on, allowing their positions to be determined with nanometer accuracy. The fluorophore positions obtained from a series of imaging cycles were used to reconstruct the overall image. We demonstrated an imaging resolution of 20 nm. This technique can, in principle, reach molecular-scale resolution.read more
Citations
More filters
Journal ArticleDOI
Mediator and RNA polymerase II clusters associate in transcription-dependent condensates
Won-Ki Cho,Jan-Hendrik Spille,Micca Hecht,Choongman Lee,Charles H. Li,Valentin Grube,Valentin Grube,Ibrahim I Cisse +7 more
TL;DR: This work used live-cell superresolution and light-sheet imaging to study the organization and dynamics of the Mediator coactivator and RNA polymerase II (Pol II) directly and suggests that large clusters of Mediator, recruited by transcription factors at large or clustered enhancer elements, interact with large Pol II clusters in transcriptional condensates in vivo.
Journal ArticleDOI
Direct stochastic optical reconstruction microscopy with standard fluorescent probes
Sebastian van de Linde,Anna Löschberger,Teresa Klein,Meike Heidbreder,Steve Wolter,Mike Heilemann,Markus Sauer +6 more
TL;DR: A step-by-step protocol for dSTORM imaging in fixed and living cells on a wide-field fluorescence microscope, with standard fluorescent probes focusing especially on the photoinduced fine adjustment of the ratio of fluorophores residing in the ON and OFF states is presented.
Journal ArticleDOI
Interferometric fluorescent super-resolution microscopy resolves 3D cellular ultrastructure
Gleb Shtengel,James A. Galbraith,Catherine G. Galbraith,Jennifer Lippincott-Schwartz,Jennifer M. Gillette,Suliana Manley,Rachid Sougrat,Clare M. Waterman,Pakorn Kanchanawong,Michael W. Davidson,Richard D. Fetter,Harald F. Hess +11 more
TL;DR: iPALM closes the gap between electron tomography and light microscopy, enabling both molecular specification and resolution of cellular nanoarchitecture.
Journal ArticleDOI
Multiplexed 3D cellular super-resolution imaging with DNA-PAINT and Exchange-PAINT
TL;DR: The transient binding of short fluorescently labeled oligonucleotides is used for simple and easy-to-implement multiplexed super-resolution imaging that achieves sub-10-nm spatial resolution in vitro on synthetic DNA structures and a multiplexing approach (Exchange-PAINT) that allows sequential imaging of multiple targets using only a single dye and a single laser source.
Journal ArticleDOI
Transcriptome-scale super-resolved imaging in tissues by RNA seqFISH
Chee-Huat Linus Eng,Michael J. Lawson,Qian Zhu,Ruben Dries,Noushin Koulena,Yodai Takei,Jina Yun,Christopher J. Cronin,Christoph D. Karp,Guo-Cheng Yuan,Long Cai +10 more
TL;DR: It is shown that seqFISH+ can image mRNAs for 10,000 genes in single cells—with high accuracy and sub-diffraction-limit resolution—in the cortex, subventricular zone and olfactory bulb of mouse brain, using a standard confocal microscope.
References
More filters
Journal ArticleDOI
Nonlinear magic: multiphoton microscopy in the biosciences
TL;DR: Multiphoton microscopy has found a niche in the world of biological imaging as the best noninvasive means of fluorescence microscopy in tissue explants and living animals and its use is now increasing exponentially.
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.
Journal ArticleDOI
Toward fluorescence nanoscopy
TL;DR: A family of concepts has emerged that overcomes the diffraction barrier altogether and, relying on saturated optical transitions, these concepts are limited only by the attainable saturation level.
Related Papers (5)
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