A general design of caging-group-free photoactivatable fluorophores for live-cell nanoscopy

Nature Chemistry, Journal Year: 2022, Volume and Issue: 14(9), P. 1013 - 1020

Published: July 21, 2022

Abstract The controlled switching of fluorophores between non-fluorescent and fluorescent states is central to every super-resolution fluorescence microscopy (nanoscopy) technique, the exploration radically new mechanisms remains critical boosting performance established, as well emerging methods. Photoactivatable dyes offer substantial improvements many these techniques, but often rely on photolabile protecting groups that limit their applications. Here we describe a general method transform 3,6-diaminoxanthones into caging-group-free photoactivatable fluorophores. These xanthones (PaX) assemble rapidly cleanly highly fluorescent, photo- chemically stable pyronine upon irradiation with light. strategy extendable carbon- silicon-bridged xanthone analogues, yielding family labels spanning much visible spectrum. Our results demonstrate versatility utility PaX in fixed live-cell labelling for conventional microscopy, coordinate-stochastic deterministic nanoscopies STED, PALM MINFLUX.

Language: Английский

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Sensitivity optimization of a rhodopsin-based fluorescent voltage indicator

Neuron, Journal Year: 2023, Volume and Issue: 111(10), P. 1547 - 1563.e9

Published: April 3, 2023

Language: Английский

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Xanthene, cyanine, oxazine and BODIPY: the four pillars of the fluorophore empire for super-resolution bioimaging

Chemical Society Reviews, Journal Year: 2023, Volume and Issue: 52(20), P. 7197 - 7261

Published: Jan. 1, 2023

In the realm of biological research, invention super-resolution microscopy (SRM) has enabled visualization ultrafine sub-cellular structures and their functions in live cells at nano-scale level, beyond diffraction limit, which opened up a new window for advanced biomedical studies to unravel complex unknown details physiological disorders level with unprecedented resolution clarity. However, most SRM techniques are highly reliant on personalized special photophysical features fluorophores. recent times, there been an surge development robust fluorophore systems various imaging techniques. To date, xanthene, cyanine, oxazine BODIPY cores have authoritatively utilized as basic units small-molecule-based organic fluorescent probe designing strategies owing excellent characteristics easy synthetic acquiescence. Since future next-generation will be decided by availability probes these four building blocks play important role progressive design, is urgent need review advancements fluorophores different methods based dye cores. This article not only includes comprehensive discussion about developments emphasis effective integration into cell bio-imaging applications but also critically evaluates background each highlight merits demerits towards developing newer SRM.

Language: Английский

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Chemical Approaches to Optimize the Properties of Organic Fluorophores for Imaging and Sensing

Angewandte Chemie International Edition, Journal Year: 2023, Volume and Issue: 63(11)

Published: Dec. 12, 2023

Abstract Organic fluorophores are indispensable tools in cells, tissue and vivo imaging, have enabled much progress the wide range of biological biomedical fields. However, many available dyes suffer from insufficient performances, such as short absorption emission wavelength, low brightness, poor stability, small Stokes shift, unsuitable permeability, restricting their application advanced imaging technology complex imaging. Over past two decades, efforts been made to improve these performances fluorophores. Starting with luminescence principle fluorophores, this review clarifies mechanisms performance for traditional a certain extent, systematically summarizes modified approaches optimizing properties, highlights typical applications improved sensing, indicates existing problems challenges area. This not only proves significance improving but also provide theoretical guidance development high‐performance

Language: Английский

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Optimized Red-Absorbing Dyes for Imaging and Sensing

Journal of the American Chemical Society, Journal Year: 2023, Volume and Issue: 145(42), P. 23000 - 23013

Published: Oct. 16, 2023

Rhodamine dyes are excellent scaffolds for developing a broad range of fluorescent probes. A key property rhodamines is their equilibrium between colorless lactone and zwitterion. Tuning the lactone–zwitterion constant (KL–Z) can optimize dye properties specific biological applications. Here, we use known novel organic chemistry to prepare comprehensive collection rhodamine elucidate structure–activity relationships that govern KL–Z. We discovered auxochrome substituent strongly affects equilibrium, providing roadmap rational design improved dyes. Electron-donating auxochromes, such as julolidine, work in tandem with fluorinated pendant phenyl rings yield bright, red-shifted fluorophores live-cell single-particle tracking (SPT) multicolor imaging. The N-aryl combined fluorination yields Förster resonance energy transfer (FRET) quencher useful creating new semisynthetic indicator sense cAMP using fluorescence lifetime imaging microscopy (FLIM). Together, this expands synthetic methods available synthesis, generates reagents advanced experiments, describes will guide future

Language: Английский

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Strategies to convert organic fluorophores into red/near-infrared emitting analogues and their utilization in bioimaging probes

Chemical Society Reviews, Journal Year: 2023, Volume and Issue: 52(18), P. 6344 - 6358

Published: Jan. 1, 2023

This review analyzes strategies to convert key fluorophores into red/NIR emitting derivatives, with typical examples of how such can be used develop molecular probes for biological analytes, along sensing features.

Language: Английский

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Silicon-RosIndolizine fluorophores with shortwave infrared absorption and emission profiles enable in vivo fluorescence imaging

Nature Chemistry, Journal Year: 2024, Volume and Issue: 16(6), P. 970 - 978

Published: March 25, 2024

Language: Английский

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Recording physiological history of cells with chemical labeling

Science, Journal Year: 2024, Volume and Issue: 383(6685), P. 890 - 897

Published: Feb. 22, 2024

Recordings of the physiological history cells provide insights into biological processes, yet obtaining such recordings is a challenge. To address this, we introduce method to record transient cellular events for later analysis. We designed proteins that become labeled in presence both specific activity and fluorescent substrate. The recording period set by substrate, whereas controls degree labeling. use distinguishable substrates enabled successive periods activity. recorded protein-protein interactions, G protein-coupled receptor activation, increases intracellular calcium. elevated calcium levels allowed selections from heterogeneous populations transcriptomic analysis tracking neuronal activities flies zebrafish.

Language: Английский

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Molecular Engineering of Direct Activated NIR-II Chemiluminescence Platform for In Vivo Chemiluminescence-fluorescence Duplex Imaging

Nature Communications, Journal Year: 2025, Volume and Issue: 16(1)

Published: Jan. 2, 2025

Chemiluminescence (CL) is a self-illuminating phenomenon fueled by chemical energy instead of extra excited light, which features superiority in sensitivity, signal-to-background ratios, and imaging depth. Strategies to synthesize CL emission unimolecular skeleton the second near-infrared window (NIR-II) probe with direct duplex NIR-II [CL/fluorescence (FL)] are lacking. Here, we employ modular synthesis routes construct series directly activated probes maximum wavelength up 1060 nm, use them for real-time continuous detection superoxide anion generated acetaminophen induced liver injury female mice model under both FL channels. Thus, this study establishes activatable skeleton, validating scalability CL/FL platform bioactive molecule disease diagnosis. chemiluminescence authors NIRII

Language: Английский

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Advances in Chromatin Imaging at Kilobase-Scale Resolution

Trends in Genetics, Journal Year: 2020, Volume and Issue: 36(4), P. 273 - 287

Published: Jan. 29, 2020

Microscopy now allows the measurement of polymeric structure chromatin in intact, fixed nuclei, with a resolution few kilobases.Multiway contact interactions and physical separation domains can be probed directly.Structural information combined measurements nascent transcription, nuclear position, cellular morphology.Single-cell imaging validates many features previously identified by bulk, proximity-based approaches, challenges existing models structure.In situ super-resolution offers new potential for multiomic single-cell analyses. It is widely appreciated that spatial organization genome nonrandom, its complex 3D folding has important consequences processes. Recent developments multiplexed, microscopy have enabled an unprecedented view – from loose folds whole chromosomes to detailed loops cis-regulatory elements regulate gene expression. Facilitated use robotics, microfluidics, improved approaches super-resolution, thousands hundreds individual cells analyzed experiment. 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Language: Английский

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