scholarly journals Stochastic optical reconstruction microscopy (STORM) in comparison with stimulated emission depletion (STED) and other imaging methods

2015 ◽  
Vol 135 (4) ◽  
pp. 643-658 ◽  
Author(s):  
Johnny Tam ◽  
David Merino
Keyword(s):  
Stimulated Emission ◽  
Imaging Methods ◽  
Stochastic Optical Reconstruction Microscopy ◽  
Stimulated Emission Depletion ◽  
Optical Reconstruction

scholarly journals Active and inactive β1 integrins segregate into distinct nanoclusters in focal adhesions

2018 ◽  
Vol 217 (6) ◽  
pp. 1929-1940 ◽  
Author(s):  
Matthias Spiess ◽  
Pablo Hernandez-Varas ◽  
Anna Oddone ◽  
Helene Olofsson ◽  
Hans Blom ◽  
...  
Keyword(s):  
Spatial Organization ◽  
Focal Adhesions ◽  
Stimulated Emission ◽  
Superresolution Microscopy ◽  
Cell Matrix ◽  
Stochastic Optical Reconstruction Microscopy ◽  
Β1 Integrins ◽  
Activity State ◽  
Optical Reconstruction ◽  
Microscopy Techniques

Integrins are the core constituents of cell–matrix adhesion complexes such as focal adhesions (FAs) and play key roles in physiology and disease. Integrins fluctuate between active and inactive conformations, yet whether the activity state influences the spatial organization of integrins within FAs has remained unclear. In this study, we address this question and also ask whether integrin activity may be regulated either independently for each integrin molecule or through locally coordinated mechanisms. We used two distinct superresolution microscopy techniques, stochastic optical reconstruction microscopy (STORM) and stimulated emission depletion microscopy (STED), to visualize active versus inactive β1 integrins. We first reveal a spatial hierarchy of integrin organization with integrin molecules arranged in nanoclusters, which align to form linear substructures that in turn build FAs. Remarkably, within FAs, active and inactive β1 integrins segregate into distinct nanoclusters, with active integrin nanoclusters being more organized. This unexpected segregation indicates synchronization of integrin activities within nanoclusters, implying the existence of a coordinate mechanism of integrin activity regulation.

scholarly journals Stimulated Emission Depletion Microscopy with Color Centers in Hexagonal Boron Nitride

ACS Photonics ◽  
2021 ◽  
Author(s):  
Prince Khatri ◽  
Ralph Nicholas Edward Malein ◽  
Andrew J. Ramsay ◽  
Isaac J. Luxmoore
Keyword(s):  
Boron Nitride ◽  
Hexagonal Boron Nitride ◽  
Stimulated Emission ◽  
Color Centers ◽  
Stimulated Emission Depletion

scholarly journals Blinking statistics and molecular counting in direct stochastic reconstruction microscopy (dSTORM)

2021 ◽  
Author(s):  
Lekha Patel ◽  
David Williamson ◽  
Dylan M Owen ◽  
Edward A K Cohen
Keyword(s):  
Probability Distribution ◽  
Single Molecule ◽  
Immunological Synapse ◽  
Training Data ◽  
Supplementary Information ◽  
Cellular Structures ◽  
Exact Probability ◽  
Stochastic Optical Reconstruction Microscopy ◽  
Exact Probability Distribution ◽  
Optical Reconstruction

Abstract Motivation Many recent advancements in single-molecule localization microscopy exploit the stochastic photoswitching of fluorophores to reveal complex cellular structures beyond the classical diffraction limit. However, this same stochasticity makes counting the number of molecules to high precision extremely challenging, preventing key insight into the cellular structures and processes under observation. Results Modelling the photoswitching behaviour of a fluorophore as an unobserved continuous time Markov process transitioning between a single fluorescent and multiple dark states, and fully mitigating for missed blinks and false positives, we present a method for computing the exact probability distribution for the number of observed localizations from a single photoswitching fluorophore. This is then extended to provide the probability distribution for the number of localizations in a direct stochastic optical reconstruction microscopy experiment involving an arbitrary number of molecules. We demonstrate that when training data are available to estimate photoswitching rates, the unknown number of molecules can be accurately recovered from the posterior mode of the number of molecules given the number of localizations. Finally, we demonstrate the method on experimental data by quantifying the number of adapter protein linker for activation of T cells on the cell surface of the T-cell immunological synapse. Availability and implementation Software and data available at https://github.com/lp1611/mol_count_dstorm. Supplementary information Supplementary data are available at Bioinformatics online.

scholarly journals Molecular organization of mammalian meiotic chromosome axis revealed by expansion STORM microscopy

2019 ◽  
Vol 116 (37) ◽  
pp. 18423-18428 ◽  
Author(s):  
Huizhong Xu ◽  
Zhisong Tong ◽  
Qing Ye ◽  
Tengqian Sun ◽  
Zhenmin Hong ◽  
...  
Keyword(s):  
Meiotic Chromosome ◽  
Molecular Organization ◽  
Homologous Chromosomes ◽  
Meiotic Chromosomes ◽  
C Terminus ◽  
Stochastic Optical Reconstruction Microscopy ◽  
Optical Reconstruction ◽  
20 Nm ◽  
Chromosome Axis

During prophase I of meiosis, chromosomes become organized as loop arrays around the proteinaceous chromosome axis. As homologous chromosomes physically pair and recombine, the chromosome axis is integrated into the tripartite synaptonemal complex (SC) as this structure’s lateral elements (LEs). While the components of the mammalian chromosome axis/LE—including meiosis-specific cohesin complexes, the axial element proteins SYCP3 and SYCP2, and the HORMA domain proteins HORMAD1 and HORMAD2—are known, the molecular organization of these components within the axis is poorly understood. Here, using expansion microscopy coupled with 2-color stochastic optical reconstruction microscopy (STORM) imaging (ExSTORM), we address these issues in mouse spermatocytes at a resolution of 10 to 20 nm. Our data show that SYCP3 and the SYCP2 C terminus, which are known to form filaments in vitro, form a compact core around which cohesin complexes, HORMADs, and the N terminus of SYCP2 are arrayed. Overall, our study provides a detailed structural view of the meiotic chromosome axis, a key organizational and regulatory component of meiotic chromosomes.

scholarly journals Mapping Neuronal Connectivity Using Stochastic Optical Reconstruction Microscopy (Storm): The Brainstorm Project

2010 ◽  
Vol 98 (3) ◽  
pp. 214a
Author(s):  
Melike Lakadamyali ◽  
Mark Bates ◽  
Hazen Babcock ◽  
Jeff Lichtman ◽  
Xiaowei Zhuang
Keyword(s):  
Neuronal Connectivity ◽  
Stochastic Optical Reconstruction Microscopy ◽  
Optical Reconstruction
Sign in / Sign up

Export Citation Format

Share Document