Fluorescent dyes and probes for super-resolution microscopy of microtubules and tracheoles in living cells and tissues

Chemical Science
Gražvydas LukinavičiusStefan W Hell

Abstract

We introduce fluorogenic tubulin probes based on the recently reported fluorescent dyes (510R, 580CP, GeR and SiR) and chemotherapy agents - taxanes (docetaxel, cabazitaxel and larotaxel). The cytotoxicity of the final probe, its staining performance and specificity strongly depend on both components. We found correlation between the aggregation efficiency (related to the spirolactonization of fluorophore) and cytotoxicity. Probe optimization allowed us to reach 29 ± 11 nm resolution in stimulated emission depletion (STED) microscopy images of the microtubule network in living human fibroblasts. Application to living fruit fly (Drosophila melanogaster) tissues highlighted two distinct structures: microtubules and tracheoles. We identified 6-carboxy isomers of 580CP and SiR dyes as markers for chitin-containing taenidia, a component of tracheoles. STED microscopy revealed correlation between the taenidia periodicity and the diameter of the tracheole. Combined tubulin and taenidia STED imaging showed close interaction between the microtubules and respiratory networks in living tissues of the insect larvae.

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Citations

Aug 8, 2018·Proceedings of the National Academy of Sciences of the United States of America·Jennifer-Magdalena MaschStefan W Hell
Dec 13, 2018·Organic & Biomolecular Chemistry·Eszter Kozma, Péter Kele
May 24, 2019·International Journal of Molecular Sciences·Mohamed H AliMarwan Emara
May 3, 2019·Chembiochem : a European Journal of Chemical Biology·Florian GrimmVladimir N Belov
Sep 12, 2019·Frontiers in Synaptic Neuroscience·Sofiia Reshetniak, Silvio O Rizzoli
Apr 3, 2020·Organic & Biomolecular Chemistry·Rūta GerasimaitėGraŽvydas Lukinavičius
Sep 12, 2020·Proceedings of the National Academy of Sciences of the United States of America·Avik K PatiScott C Blanchard
Oct 15, 2019·Molecular Biology of the Cell·Glenn CarringtonMichelle Peckham
Jan 15, 2020·Nature Communications·Mikhail E MatlashovVladislav V Verkhusha
Mar 3, 2020·Nanoscale Horizons : the Home for Rapid Reports of Exceptional Significance in Nanoscience and Nanotechnolgy·Qiang WuHongda Wang
Mar 19, 2019·Chemical Science·Jonas BucevičiusGražvydas Lukinavičius
Dec 20, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Sascha G KellerYasuteru Urano
Jan 7, 2021·Organic & Biomolecular Chemistry·Veselin NasufovićHans-Dieter Arndt
Jan 27, 2021·Chemistry : a European Journal·Florian GrimmStefan W Hell
Mar 30, 2021·Chemical Science·Jonas BucevičiusGražvydas Lukinavičius
Jul 11, 2021·Bioorganic & Medicinal Chemistry Letters·Shang Jia, Christopher J Chang
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Oct 1, 2021·Chemical Society Reviews·Jana VolarićBen L Feringa
Nov 5, 2021·ACS Chemical Biology·Ru Ta GerasimaitėGražvydas Lukinavičius

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Methods Mentioned

BETA
super-resolution microscopy
light microscopy
confocal microscopy
light scattering
electron microscopy
Fluorescence

Software Mentioned

ChemDraw Professional
AutoDock Tools
Fiji
ImSpector
GraphPad Prism
AutoDock Vina
Vina Autodock
ChemoMetec NucleoView NC - 3000

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