Imagining the future of optical microscopy: everything, everywhere, all at once

Communications Biology, Год журнала: 2023, Номер 6(1)

Опубликована: Окт. 28, 2023

The optical microscope has revolutionized biology since at least the 17

Язык: Английский

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Chemical Control of Fluorescence Lifetime towards Multiplexing Imaging

Angewandte Chemie International Edition, Год журнала: 2024, Номер 63(25)

Опубликована: Апрель 20, 2024

Abstract Fluorescence lifetime imaging has been a powerful tool for biomedical research. Recently, fluorescence lifetime‐based multiplexing expanded channels by using probes that harbor the same spectral and distinct excited state lifetime. While it is desirable to control of any given fluorescent probes, rational lifetimes remains challenge. Herein, we chose boron dipyrromethene (BODIPY) as model system provided chemical strategies regulate its derivatives with varying features. We find electronegativity structural substituents at 8′ 5′ positions important green‐emitting red‐emitting BODIPY scaffolds. Mechanistically, such influences are exerted via photo‐induced electron transfer intramolecular charge processes BODIPY, respectively. Based on these principles, have generated group enable experiments separate multiple targets signal. In addition envision modulation could serve feasible strategy achieve variety small molecule fluorophores.

Язык: Английский

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A guide to single-particle tracking

Nature Reviews Methods Primers, Год журнала: 2024, Номер 4(1)

Опубликована: Сен. 12, 2024

Язык: Английский

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Harnessing the Power of Plasmonics for in Vitro and in Vivo Biosensing

ACS Photonics, Год журнала: 2025, Номер unknown

Опубликована: Фев. 17, 2025

Plasmonic nanostructures exhibit localized surface plasmon resonances due to collective oscillation of conducting electrons that can be tuned by modulating the nanostructure size, shape, material composition, and local dielectric environment. The strong field confinement enhancement provided plasmonic have been exploited over years enhance sensitivity for analyte detection down single-molecule level, rendering these devices as potentially outstanding biosensors. Here, we summarize methods detect biological analytes in vitro living cells, with a focus on plasmon-enhanced fluorescence, Raman scattering, infrared absorption, circular dichroism, refractive index sensing. Given tremendous advances field, concentrate few recent examples toward biosensing under highly challenging conditions, including clinically relevant biomarkers body fluids nascent applications cells vivo. These emerging platforms serve inspiration exploring future directions nanoplasmonics further harnessed advance real-world applications.

Язык: Английский

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Fluorogenic Cyclopropenones for Multicomponent, Real-Time Imaging

Journal of the American Chemical Society, Год журнала: 2022, Номер 144(17), С. 7871 - 7880

Опубликована: Апрель 20, 2022

Fluorogenic bioorthogonal reactions enable biomolecule visualization in real time. These comprise reporters that "light up" upon reaction with complementary partners. While the spectrum of fluorogenic chemistries is expanding, few transformations are compatible live cells due to cross-reactivities or insufficient signal turn-on. To address need for more suitable cellular imaging, we developed a featuring cyclopropenone and phosphines. The transformation involves regioselective activation cyclization cyclopropenones form coumarin products. With optimal probes, provides >1600-fold turn-on, one highest fluorescence enhancements reported date. motifs were evaluated vitro cells. was also found be other common transformations, enabling multicomponent, real-time imaging. Collectively, these data suggest cyclopropenone–phosphine will bolster efforts track targets their native settings.

Язык: Английский

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Imaging plant cells and organs with light-sheet and super-resolution microscopy

PLANT PHYSIOLOGY, Год журнала: 2021, Номер 188(2), С. 683 - 702

Опубликована: Июль 21, 2021

Abstract The documentation of plant growth and development requires integrative scalable approaches to investigate spatiotemporally resolve various dynamic processes at different levels body organization. present update deals with vigorous developments in mesoscopy, microscopy nanoscopy methods that have been translated imaging subcellular compartments, cells, tissues organs over the past 3 years aim report recent applications reasonable expectations from current light-sheet fluorescence (LSFM) super-resolution (SRM) modalities. Moreover, shortcomings limitations existing LSFM SRM are discussed, particularly for their ability accommodate samples regarding potential considering spherical aberrations or temporal restrictions prohibiting recording fast cellular three dimensions. For a more comprehensive description, advances living fixed sample preparation also included, supported by an overview labeling strategies successfully applied plants. These practically documented employing model Arabidopsis thaliana (L.) Heynh., but robust crop species such as Medicago sativa L. Hordeum vulgare Over few years, trend towards designing microscopic modalities has become apparent it is expected near future will be bridged achieve broader multiscale single platform.

Язык: Английский

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Genetically encodable fluorescent protein markers in advanced optical imaging

Methods and Applications in Fluorescence, Год журнала: 2022, Номер 10(4), С. 042002 - 042002

Опубликована: Июнь 29, 2022

Abstract Optical fluorescence microscopy plays a pivotal role in the exploration of biological structure and dynamics, especially on live specimens. Progress field relies, one hand, technical advances imaging data processing and, other progress fluorescent marker technologies. Among these, genetically encodable proteins (FPs) are invaluable tools, as they allow facile labeling cells, tissues or organisms, these produce FP markers all by themselves after introduction suitable gene. Here we cover from GFP family well tetrapyrrole-binding proteins, which further complement toolbox important ways. A broad range variants have been endowed, using protein engineering, with photophysical properties that essential for specific techniques, notably those offering nanoscale image resolution. We briefly introduce various advanced methods show how utilize distinct exciting applications, aim to guide researchers toward design powerful experiments optimally suited address their questions.

Язык: Английский

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Labeling of heterochronic ribosomes reveals C1ORF109 and SPATA5 control a late step in human ribosome assembly

Cell Reports, Год журнала: 2022, Номер 38(13), С. 110597 - 110597

Опубликована: Март 1, 2022

Although features of ribosome assembly are shared between species, our understanding the diversity, complexity, dynamics, and regulation production in multicellular organisms remains incomplete. To gain insights into biogenesis human cells, we perform a genome-wide loss-of-function screen combined with differential labeling pre-existing newly assembled ribosomes. These efforts identify two functionally uncharacterized genes, C1orf109 SPATA5. We provide evidence that these factors, together CINP SPATA5L1, control late step pre-60S maturation cytoplasm. Loss either or SPATA5 impairs global protein synthesis. results link neurodevelopmental disorders associated recessive mutations. Based on findings, propose expanded repertoire factors likely enables to coordinate multiple steps response different developmental environmental stimuli.

Язык: Английский

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Design of a palette of SNAP-tag mimics of fluorescent proteins and their use as cell reporters

Cell Discovery, Год журнала: 2023, Номер 9(1)

Опубликована: Июнь 13, 2023

Naturally occurring fluorescent proteins (FPs) are the most widely used tools for tracking cellular and sensing events. Here, we chemically evolved self-labeling SNAP-tag into a palette of mimics (SmFPs) that possess bright, rapidly inducible fluorescence ranging from cyan to infrared. SmFPs integral chemical-genetic entities based on same fluorogenic principle as FPs, i.e., induction non-emitting molecular rotors by conformational locking. We demonstrate usefulness these in real-time protein expression, degradation, binding interactions, trafficking, assembly, show optimally designed outperform FPs like GFP many important ways. further circularly permuted is sensitive changes their fusion partners, partners can be development single SmFP-based genetically encoded calcium sensors live cell imaging.

Язык: Английский

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Genetic Targeting of Solvatochromic Dyes for Probing Nanoscale Environments of Proteins in Organelles

Analytical Chemistry, Год журнала: 2023, Номер 95(22), С. 8512 - 8521

Опубликована: Май 25, 2023

A variety of protein tags are available for genetically encoded labeling, which allow their precise localization and tracking inside the cells. new dimension in imaging can be offered by combining with polarity-sensitive fluorescent probes, provide information about local nanoscale environments target proteins within subcellular compartments (organelles). Here, we designed three probes based on solvatochromic nile red dye, conjugated to a HaloTag reactive targeting group through polyethylene glycol linkers varying lengths. The probe medium linker length, NR12-Halo, was found label specifically large localized defined cell compartments, such as plasma membranes (outer inner leaflets), endoplasmic reticulum, Golgi apparatus, cytosol, microtubules, actin, chromatin. Owing its fluorophore, clearly distinguished apolar lipid from other proteins. Moreover, it revealed dramatic changes environment during life cycle biosynthesis expected and, finally, recycling lysosomes. Heterogeneity polarity some membrane also suggested formation low-polar aggregates, example, cell–cell contacts. approach showed that mechanical stress (cell shrinking osmotic shock) induced general decrease proteins, probably due condensation biomolecules. Finally, nanoenvironment affected polyunsaturated fatty acid diet, provided bridge between organization lipids developed constitutes promising tool probing interactions structures.

Язык: Английский

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