Subdiffraction-resolution fluorescence microscopy reveals a domain of the centrosome critical for pericentriolar material organization
Vito Mennella,Bettina Keszthelyi,Kent L. McDonald,Bryant B. Chhun,F Kan,Gregory C. Rogers,Bo Huang,David A. Agard +7 more
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TLDR
It is demonstrated that the pericentriolar material is organized into two main structural domains: a layer juxtaposed to the centriole wall, and proteins extending farther away from the Centrosome organized in a matrix, using SIM and STORM subdiffraction-resolution microscopies.Abstract:
Centrosomes, the microtubule nucleation centre of most cells, consist of two centrioles surrounded by pericentriolar material (PCM). The PCM has been considered as amorphous but, using subdiffraction fluorescence microscopy approaches, Agard and colleagues now reveal the organized structure of the PCM of Drosophila centrosomes.read more
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Super-resolution microscopy demystified
Lothar Schermelleh,Alexia Ferrand,Thomas R Huser,Christian Eggeling,Markus Sauer,Oliver Biehlmaier,Drummen Gpc. +6 more
TL;DR: An overview of current super-resolution microscopy techniques is given and guidance on how best to use them to foster biological discovery is provided.
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The Centrosome Is a Selective Condensate that Nucleates Microtubules by Concentrating Tubulin
Jeffrey B. Woodruff,Beatriz Ferreira Gomes,Per O. Widlund,Julia Mahamid,Alf Honigmann,Anthony A. Hyman +5 more
TL;DR: It is found that macromolecular crowding drives assembly of the key PCM scaffold protein SPD-5 into spherical condensates that morphologically and dynamically resemble in vivo PCM.
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TL;DR: A lucid synthesis of the developments on single-molecule localization precision and accuracy and their practical implications are presented in order to guide the increasing number of researchers using single-particle tracking and super-resolution localization microscopy.
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Centrosome function and assembly in animal cells
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References
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Journal ArticleDOI
Microtubule nucleation by γ-tubulin complexes
TL;DR: The first crystallographic analysis of a non-γ-tubulin γTuRC component has resulted in a new appreciation of the relationships among all γ TuRC proteins, leading to a refined model of their organization and function.
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Three-Dimensional Super-Resolution Imaging by Stochastic Optical Reconstruction Microscopy
TL;DR: 3D stochastic optical reconstruction microscopy (STORM) is demonstrated by using optical astigmatism to determine both axial and lateral positions of individual fluorophores with nanometer accuracy, allowing the 3D morphology of nanoscopic cellular structures to be resolved.
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Super-Resolution Fluorescence Microscopy
TL;DR: It is anticipated that super-resolution fluorescence microscopy will become a widely used tool for cell and tissue imaging to provide previously unobserved details of biological structures and processes.
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Three-Dimensional Resolution Doubling in Wide-Field Fluorescence Microscopy by Structured Illumination
Mats G. L. Gustafsson,Lin Shao,Peter M. Carlton,C. J. Rachel Wang,Inna N. Golubovskaya,W. Zacheus Cande,David A. Agard,David A. Agard,John W. Sedat +8 more
TL;DR: This work describes how spatially structured illumination microscopy can be applied in three dimensions to double the axial as well as the lateral resolution, with true optical sectioning, and has produced the first light microscopy images of the synaptonemal complex in which the lateral elements are clearly resolved.
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Proteomic characterization of the human centrosome by protein correlation profiling
Jens S. Andersen,Christopher J. Wilkinson,Thibault Mayor,Thibault Mayor,Peter Mortensen,Erich A. Nigg,Matthias Mann +6 more
TL;DR: A mass-spectrometry-based proteomic analysis of human centrosomes in the interphase of the cell cycle by quantitatively profiling hundreds of proteins across several centrifugation fractions identified and validated 23 novel components and identified 41 likely candidates as well as the vast majority of the known centrosomal proteins in a large background of nonspecific proteins.