High-speed Live-cell Super-resolution Microscopy with Stochastically Switching Fluorophores

CLEO: 2013 ◽  
2013 ◽  
Author(s):  
Fang Huang ◽  
Tobias M. P. Hartwich ◽  
Felix E. Rivera-Molina ◽  
Yu Lin ◽  
Jordan R. Myers ◽  
...  
Keyword(s):  
High Speed ◽  
Super Resolution ◽  
Live Cell ◽  
Super Resolution Microscopy

scholarly journals Advances in High-Speed Structured Illumination Microscopy

2021 ◽  
Vol 9 ◽  
Author(s):  
Tianyu Zhao ◽  
Zhaojun Wang ◽  
Tongsheng Chen ◽  
Ming Lei ◽  
Baoli Yao ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
High Speed ◽  
Cell Imaging ◽  
Super Resolution ◽  
Live Cell ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Light Power ◽  
Recent Developments ◽  
Super Resolution Microscopy

Super-resolution microscopy surpasses the diffraction limit to enable the observation of the fine details in sub-cellular structures and their dynamics in diverse biological processes within living cells. Structured illumination microscopy (SIM) uses a relatively low illumination light power compared with other super-resolution microscopies and has great potential to meet the demands of live-cell imaging. However, the imaging acquisition and reconstruction speeds limit its further applications. In this article, recent developments all targeted at improving the overall speed of SIM are reviewed. These comprise both hardware and software improvements, which include a reduction in the number of raw images, GPU acceleration, deep learning and the spatial domain reconstruction. We also discuss the application of these developments in live-cell imaging.

scholarly journals Reticulon and CLIMP-63 control nanodomain organization of peripheral ER tubules

2019 ◽  
Author(s):  
Guang Gao ◽  
Chengjia Zhu ◽  
Emma Liu ◽  
Ivan R. Nabi
Keyword(s):  
Endoplasmic Reticulum ◽  
Confocal Microscopy ◽  
High Speed ◽  
Super Resolution ◽  
Live Cell ◽  
Stimulated Emission ◽  
Preferential Segregation ◽  
Stimulated Emission Depletion ◽  
Super Resolution Microscopy ◽  
Local Accumulation

AbstractThe endoplasmic reticulum (ER) is an expansive, membrane-enclosed organelle composed of smooth peripheral tubules and rough, ribosome-studded central ER sheets whose morphology is determined, in part, by the ER-shaping proteins, reticulon and CLIMP-63, respectively. Here, STimulated Emission Depletion (STED) super-resolution microscopy shows that reticulon and CLIMP-63 also control the organization and dynamics of peripheral ER tubule nanodomains. STED imaging shows that lumenal ERmoxGFP, membrane Sec61βGFP, knock-in calreticulin-GFP and antibody-labeled ER resident proteins calnexin and derlin-1 are all localized to periodic puncta along the length of peripheral ER tubules that are not readily observable by diffraction limited confocal microscopy. Reticulon segregates away from and restricts lumenal blob length while CLIMP-63 associates with and increases lumenal blob length. Reticulon and CLIMP-63 also regulate the nanodomain distribution of ER resident proteins, being required for the preferential segregation of calnexin and derlin-1 puncta away from lumenal ERmoxGFP blobs. High-speed (40 ms/frame) live cell STED imaging shows that reticulon and CLIMP-63 control nanoscale compartmentalization of lumenal flow in peripheral ER tubules. Reticulon enhances and CLIMP-63 disrupts the local accumulation of lumenal ERmoxGFP at spatially defined sites along ER tubules. The ER shaping proteins reticulon and CLIMP-63 therefore control lumenal ER nanodomain dynamics, heterogeneity and interaction with ER resident proteins in peripheral ER tubules.

scholarly journals Bio-orthogonal Red and Far-Red Fluorogenic Probes for Wash-Free Live-Cell and Super-resolution Microscopy

2021 ◽  
Author(s):  
Philipp Werther ◽  
Klaus Yserentant ◽  
Felix Braun ◽  
Kristin Grußmayer ◽  
Vytautas Navikas ◽  
...  
Keyword(s):  
Super Resolution ◽  
Live Cell ◽  
Fluorogenic Probes ◽  
Super Resolution Microscopy

scholarly journals Ultrasmall, Bright, and Photostable Fluorescent Core–Shell Aluminosilicate Nanoparticles for Live‐Cell Optical Super‐Resolution Microscopy

2021 ◽  
Vol 33 (8) ◽  
pp. 2006829
Author(s):  
Jacob A. Erstling ◽  
Joshua A. Hinckley ◽  
Nirmalya Bag ◽  
Jessica Hersh ◽  
Grant B. Feuer ◽  
...  
Keyword(s):  
Super Resolution ◽  
Live Cell ◽  
Core Shell ◽  
Super Resolution Microscopy

scholarly journals Live cell single molecule-guided Bayesian localization super resolution microscopy

Cell Research ◽  
2016 ◽  
Vol 27 (5) ◽  
pp. 713-716 ◽  
Author(s):  
Fan Xu ◽  
Mingshu Zhang ◽  
Wenting He ◽  
Renmin Han ◽  
Fudong Xue ◽  
...  
Keyword(s):  
Single Molecule ◽  
Super Resolution ◽  
Live Cell ◽  
Super Resolution Microscopy

Small-Molecule Fluorescent Probes for Live-Cell Super-Resolution Microscopy

2018 ◽  
Vol 141 (7) ◽  
pp. 2770-2781 ◽  
Author(s):  
Lu Wang ◽  
Michelle S. Frei ◽  
Aleksandar Salim ◽  
Kai Johnsson
Keyword(s):  
Small Molecule ◽  
Fluorescent Probes ◽  
Super Resolution ◽  
Live Cell ◽  
Super Resolution Microscopy
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