scholarly journals Combined Use of Unnatural Amino Acids Enables Dual-Color Super-Resolution Imaging of Proteins via Click Chemistry

ACS Nano ◽  
2018 ◽  
Vol 12 (12) ◽  
pp. 12247-12254 ◽  
Author(s):  
Kim-A. Saal ◽  
Frank Richter ◽  
Peter Rehling ◽  
Silvio O. Rizzoli
Keyword(s):  
Amino Acids ◽  
Click Chemistry ◽  
Unnatural Amino Acids ◽  
Super Resolution ◽  
Dual Color ◽  
Combined Use ◽  
Resolution Imaging

Dual-color super-resolution imaging of quantum dot clusters

2020 ◽  
Author(s):  
Duncan Ryan ◽  
Megan K. Dunlap ◽  
Somak Majumder ◽  
Chris J. Sheehan ◽  
James H. Werner ◽  
...  
Keyword(s):  
Quantum Dot ◽  
Super Resolution ◽  
Dual Color ◽  
Resolution Imaging

scholarly journals Bioorthogonal labeling of transmembrane proteins with non-canonical amino acids allows access to masked epitopes in live neurons

2021 ◽  
Author(s):  
Diogo Bessa-Neto ◽  
Alexander Kuhlemann ◽  
Gerti Beliu ◽  
Valeria Pecoraro ◽  
Sören Doose ◽  
...  
Keyword(s):  
Amino Acids ◽  
Brain Slices ◽  
Transmembrane Proteins ◽  
Super Resolution ◽  
Biological Imaging ◽  
Bioorthogonal Labeling ◽  
Resolution Imaging ◽  
Genetic Code Expansion ◽  
Super Resolution Microscopy ◽  
Organotypic Brain Slices

ABSTRACTProgress in biological imaging is intrinsically linked to advances in labeling methods. The explosion in the development of high-resolution and super-resolution imaging calls for new approaches to label targets with small probes. These should allow to faithfully report the localization of the target within the imaging resolution – typically nowadays a few nanometers - and allow access to any epitope of the target, in the native cellular and tissue environment. We report here the development of a complete labeling and imaging pipeline using genetic code expansion and non-canonical amino acids in primary neurons that allows to fluorescently label masked epitopes in target transmembrane proteins in live neurons, both in dissociated culture and organotypic brain slices. This allowed us to image the differential localization of two glutamate receptor auxiliary proteins in complex with their partner with a variety of methods including widefield, confocal, and dSTORM super-resolution microscopy.

Bioorthogonal Click Chemistry Enables Site-specific Fluorescence Labeling of Functional NMDA Receptors for Super-Resolution Imaging

2018 ◽  
Vol 130 (50) ◽  
pp. 16602-16607 ◽  
Author(s):  
Franziska Neubert ◽  
Gerti Beliu ◽  
Ulrich Terpitz ◽  
Christian Werner ◽  
Christian Geis ◽  
...  
Keyword(s):  
Nmda Receptors ◽  
Click Chemistry ◽  
Super Resolution ◽  
Fluorescence Labeling ◽  
Site Specific ◽  
Specific Fluorescence ◽  
Resolution Imaging

scholarly journals Dual-color super-resolution imaging by fluorescence emission depletion (STED) microscopy

2017 ◽  
Vol 844 ◽  
pp. 012033
Author(s):  
Wen-sheng Wang ◽  
Cui-fang Kuang ◽  
Shao-cong Liu ◽  
Shi-yi Sun ◽  
Xu Liu
Keyword(s):  
Fluorescence Emission ◽  
Super Resolution ◽  
Sted Microscopy ◽  
Dual Color ◽  
Resolution Imaging

Synthesis of Threefold Glycosylated Proteins using Click Chemistry and Genetically Encoded Unnatural Amino Acids

ChemBioChem ◽  
2009 ◽  
Vol 10 (18) ◽  
pp. 2858-2861 ◽  
Author(s):  
Emine Kaya ◽  
Katrin Gutsmiedl ◽  
Milan Vrabel ◽  
Markus Müller ◽  
Peter Thumbs ◽  
...  
Keyword(s):  
Amino Acids ◽  
Click Chemistry ◽  
Unnatural Amino Acids ◽  
Glycosylated Proteins

scholarly journals Full-field dual-color 100-nm super-resolution imaging reveals organization and dynamics of mitochondrial and ER networks

2013 ◽  
Vol 21 (22) ◽  
pp. 26162 ◽  
Author(s):  
Maia Brunstein ◽  
Kai Wicker ◽  
Karine Hérault ◽  
Rainer Heintzmann ◽  
Martin Oheim
Keyword(s):  
Super Resolution ◽  
Full Field ◽  
Dual Color ◽  
Resolution Imaging

scholarly journals Minimal genetically encoded tags for fluorescent protein labeling in living neurons

2021 ◽  
Author(s):  
Aleksandra Arsić ◽  
Cathleen Hagemann ◽  
Nevena Stajković ◽  
Timm Schubert ◽  
Ivana Nikić-Spiegel
Keyword(s):  
Click Chemistry ◽  
Unnatural Amino Acids ◽  
Genome Engineering ◽  
Fluorescent Protein ◽  
Super Resolution ◽  
Live Cell ◽  
Cell Labeling ◽  
Neurofilament Light Chain ◽  
Neurofilament Light ◽  
Fixed Cell

AbstractModern light microscopy, including super-resolution techniques, brought about a demand for small labeling tags that bring the fluorophore closer to the target. This challenge can be addressed by labeling unnatural amino acids (UAAs) with click chemistry. UAAs are site-specifically incorporated into a protein of interest by genetic code expansion. If the UAA carries a strained alkene or alkyne moiety it can be conjugated to a tetrazine-bearing fluorophore via a strain-promoted inverse-electron-demand Diels–Alder cycloaddition (SPIEDAC), a variant of bioorthogonal click chemistry. The minimal size of the incorporated tag and the possibility to couple the fluorophores directly to the protein of interest with single-residue precision make SPIEDAC live-cell labeling unique. However, until now, this type of labeling has not been used in complex, non-dividing cells, such as neurons. Using neurofilament light chain as a target protein, we established SPIEDAC labeling in living primary neurons and applied it for fixed-cell, live-cell, dual-color pulse—chase and super-resolution microscopy. We also show that SPIEDAC labeling can be combined with CRISPR/Cas9 genome engineering for tagging endogenous NFL. Due to its versatile nature and compatibility with advanced microscopy techniques, we anticipate that SPIEDAC labeling will contribute to novel discoveries in neurobiology.

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