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 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 Mapping the dynamics and nanoscale organization of synaptic adhesion proteins using monomeric streptavidin

2016 ◽  
Vol 7 (1) ◽  
Author(s):  
Ingrid Chamma ◽  
Mathieu Letellier ◽  
Corey Butler ◽  
Béatrice Tessier ◽  
Kok-Hong Lim ◽  
...  
Keyword(s):  
High Resolution ◽  
Membrane Proteins ◽  
Steric Hindrance ◽  
Transmembrane Protein ◽  
Super Resolution ◽  
Leucine Rich Repeat ◽  
Versatile Tool ◽  
Resolution Imaging ◽  
Dual Colour ◽  
Specific Labelling

Abstract The advent of super-resolution imaging (SRI) has created a need for optimized labelling strategies. We present a new method relying on fluorophore-conjugated monomeric streptavidin (mSA) to label membrane proteins carrying a short, enzymatically biotinylated tag, compatible with SRI techniques including uPAINT, STED and dSTORM. We demonstrate efficient and specific labelling of target proteins in confined intercellular and organotypic tissues, with reduced steric hindrance and no crosslinking compared with multivalent probes. We use mSA to decipher the dynamics and nanoscale organization of the synaptic adhesion molecules neurexin-1β, neuroligin-1 (Nlg1) and leucine-rich-repeat transmembrane protein 2 (LRRTM2) in a dual-colour configuration with GFP nanobody, and show that these proteins are diffusionally trapped at synapses where they form apposed trans-synaptic adhesive structures. Furthermore, Nlg1 is dynamic, disperse and sensitive to synaptic stimulation, whereas LRRTM2 is organized in compact and stable nanodomains. Thus, mSA is a versatile tool to image membrane proteins at high resolution in complex live environments, providing novel information about the nano-organization of biological structures.

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