scholarly journals Visualization and Manipulation of Actin Cytoskeleton with Small-Molecular Probes

2020 ◽  
Author(s):  
Takeru Takagi ◽  
Tasuku Ueno ◽  
Keisuke Ikawa ◽  
Daisuke Asanuma ◽  
Yusuke Nomura ◽  
...  
Keyword(s):  
Actin Cytoskeleton ◽  
Green Light ◽  
Dynamic Network ◽  
Super Resolution ◽  
Molecular Probes ◽  
Actin Binding ◽  
Mechanical Forces ◽  
Cellular Functions ◽  
New Class ◽  
Resolution Imaging

Actin is a ubiquitous cytoskeletal protein, forming a dynamic network that generates mechanical forces in the cell. Here, in order to dissect the complex mechanisms of actin-related cellular functions, we introduce two powerful tools based on a new class of actin-binding small molecule: one enables visualization of the actin cytoskeleton, including super-resolution imaging, and the other enables highly specific green-light-controlled fragmentation of actin filaments, affording unprecedented control of the actin cytoskeleton and its force network in living cells.

scholarly journals Visualization and Manipulation of Actin Cytoskeleton with Small-Molecular Probes

2020 ◽  
Author(s):  
Takeru Takagi ◽  
Tasuku Ueno ◽  
Keisuke Ikawa ◽  
Daisuke Asanuma ◽  
Yusuke Nomura ◽  
...  
Keyword(s):  
Actin Cytoskeleton ◽  
Green Light ◽  
Dynamic Network ◽  
Super Resolution ◽  
Molecular Probes ◽  
Actin Binding ◽  
Mechanical Forces ◽  
Cellular Functions ◽  
New Class ◽  
Resolution Imaging

Actin is a ubiquitous cytoskeletal protein, forming a dynamic network that generates mechanical forces in the cell. Here, in order to dissect the complex mechanisms of actin-related cellular functions, we introduce two powerful tools based on a new class of actin-binding small molecule: one enables visualization of the actin cytoskeleton, including super-resolution imaging, and the other enables highly specific green-light-controlled fragmentation of actin filaments, affording unprecedented control of the actin cytoskeleton and its force network in living cells.

scholarly journals Comparing lifeact and phalloidin for super-resolution imaging of actin in fixed cells

PLoS ONE ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. e0246138
Author(s):  
Hanieh Mazloom-Farsibaf ◽  
Farzin Farzam ◽  
Mohamadreza Fazel ◽  
Michael J. Wester ◽  
Marjolein B. M. Meddens ◽  
...  
Keyword(s):  
Single Molecule ◽  
Cell Biology ◽  
Cell Movement ◽  
Super Resolution ◽  
Actin Binding ◽  
Thin Filaments ◽  
Reversible Binding ◽  
Multiple Regions ◽  
Resolution Imaging ◽  
Division Cell

Visualizing actin filaments in fixed cells is of great interest for a variety of topics in cell biology such as cell division, cell movement, and cell signaling. We investigated the possibility of replacing phalloidin, the standard reagent for super-resolution imaging of F-actin in fixed cells, with the actin binding peptide ‘lifeact’. We compared the labels for use in single molecule based super-resolution microscopy, where AlexaFluor 647 labeled phalloidin was used in a dSTORM modality and Atto 655 labeled lifeact was used in a single molecule imaging, reversible binding modality. We found that imaging with lifeact had a comparable resolution in reconstructed images and provided several advantages over phalloidin including lower costs, the ability to image multiple regions of interest on a coverslip without degradation, simplified sequential super-resolution imaging, and more continuous labeling of thin filaments.

scholarly journals Super-resolution imaging of live sperm reveals dynamic changes of the actin cytoskeleton during acrosomal exocytosis

2018 ◽  
Vol 131 (21) ◽  
pp. jcs218958 ◽  
Author(s):  
Ana Romarowski ◽  
Ángel G. Velasco Félix ◽  
Paulina Torres Rodríguez ◽  
María G. Gervasi ◽  
Xinran Xu ◽  
...  
Keyword(s):  
Actin Cytoskeleton ◽  
Super Resolution ◽  
Dynamic Changes ◽  
Acrosomal Exocytosis ◽  
Resolution Imaging ◽  
Live Sperm

scholarly journals Comparing Lifeact and Phalloidin for Super-resolution Imaging of Actin in Fixed Cells

2020 ◽  
Author(s):  
Hanieh Mazloom-Farsibaf ◽  
Farzin Farzam ◽  
Mohamadreza Fazel ◽  
Michael J. Wester ◽  
Marjolein B. M. Meddens ◽  
...  
Keyword(s):  
Single Molecule ◽  
Cell Biology ◽  
Cell Movement ◽  
Super Resolution ◽  
Actin Binding ◽  
Thin Filaments ◽  
Reversible Binding ◽  
Multiple Regions ◽  
Resolution Imaging ◽  
Division Cell

AbstractVisualizing actin filaments in fixed cells is of great interest for a variety of topics in cell biology such as cell division, cell movement, and cell signaling. We investigated the possibility of replacing phalloidin, the standard reagent for super-resolution imaging of F-actin in fixed cells, with the actin binding peptide ‘lifeact’. We compared the labels for use in single molecule based super-resolution microscopy, where AlexaFluor 647 labeled phalloidin was used in a (d)STORM modality and Atto 655 labeled lifeact was used in a single molecule imaging, reversible binding modality. We found that imaging with lifeact had a comparable resolution in reconstructed images and provided several advantages over phalloidin including lower costs, the ability to image multiple regions of interest on a coverslip without degradation, simplified sequential super-resolution imaging, and more continuous labeling of thin filaments.

A green-light-emitting, spontaneously blinking fluorophore based on intramolecular spirocyclization for dual-colour super-resolution imaging

2018 ◽  
Vol 54 (1) ◽  
pp. 102-105 ◽  
Author(s):  
Shin-nosuke Uno ◽  
Mako Kamiya ◽  
Akihiko Morozumi ◽  
Yasuteru Urano
Keyword(s):  
Green Light ◽  
Super Resolution ◽  
Light Emitting ◽  
Resolution Imaging ◽  
Dual Colour

We have developed the first green-light-emitting, spontaneously blinking fluorophore (SBF), HEtetTFER.

scholarly journals Super-Resolution Imaging by Dielectric Superlenses: TiO2 Metamaterial Superlens versus BaTiO3 Superlens

Photonics ◽  
2021 ◽  
Vol 8 (6) ◽  
pp. 222
Author(s):  
Rakesh Dhama ◽  
Bing Yan ◽  
Cristiano Palego ◽  
Zengbo Wang
Keyword(s):  
Super Resolution ◽  
Field Of View ◽  
Nano Particles ◽  
Imaging Systems ◽  
Label Free ◽  
Theoretical Simulation ◽  
New Class ◽  
Physical Shape ◽  
Resolution Imaging ◽  
First Time

All-dielectric superlens made from micro and nano particles has emerged as a simple yet effective solution to label-free, super-resolution imaging. High-index BaTiO3 Glass (BTG) microspheres are among the most widely used dielectric superlenses today but could potentially be replaced by a new class of TiO2 metamaterial (meta-TiO2) superlens made of TiO2 nanoparticles. In this work, we designed and fabricated TiO2 metamaterial superlens in full-sphere shape for the first time, which resembles BTG microsphere in terms of the physical shape, size, and effective refractive index. Super-resolution imaging performances were compared using the same sample, lighting, and imaging settings. The results show that TiO2 meta-superlens performs consistently better over BTG superlens in terms of imaging contrast, clarity, field of view, and resolution, which was further supported by theoretical simulation. This opens new possibilities in developing more powerful, robust, and reliable super-resolution lens and imaging systems.

scholarly journals Molecular organization and mechanics of single vimentin filaments revealed by super-resolution imaging

2021 ◽  
Author(s):  
Filipe Nunes Vicente ◽  
Mickael Lelek ◽  
Jean-Yves Tinevez ◽  
Quang D. Tran ◽  
Gerard Pehau-Arnaudet ◽  
...  
Keyword(s):  
Unit Length ◽  
Super Resolution ◽  
Molecular Organization ◽  
Cellular Functions ◽  
Filament Assembly ◽  
Resolution Imaging ◽  
Partial Overlap ◽  
Vimentin Filaments ◽  
In Cells

Intermediate filaments (IF) are involved in key cellular functions including polarization, migration, and protection against large deformations. These functions are related to their remarkable ability to extend without breaking, a capacity that should be determined by the molecular organization of subunits within filaments. However, this structure-mechanics relationship remains poorly understood at the molecular level. Here, using super-resolution microscopy (SRM), we show that vimentin filaments exhibit a ~49 nm axial repeat both in cells and in vitro. As unit-length-filaments (ULFs) precursors were measured at ~59 nm, this demonstrates a partial overlap of ULFs during filament assembly. Using an SRM-compatible stretching device, we also provide evidence that the extensibility of vimentin is due to the unfolding of its subunits and not to their sliding, thus establishing a direct link between the structural organization and its mechanical properties. Overall, our results pave the way for future studies of IF assembly, mechanical and structural properties in cells.

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