Phasor-FSTM: a new paradigm for multicolor super-resolution imaging of living cells based on fluorescence modulation and lifetime multiplexing DOI Creative Commons
Luwei Wang, Yue Chen,

Jiaqing Guo

et al.

Light Science & Applications, Journal Year: 2025, Volume and Issue: 14(1)

Published: Jan. 3, 2025

Abstract Multicolor microscopy and super-resolution optical are two widely used techniques that greatly enhance the ability to distinguish resolve structures in cellular imaging. These methods have individually transformed imaging by allowing detailed visualization of subcellular structures, as well organelle interactions. However, integrating multicolor into a single method remains challenging due issues like spectral overlap, crosstalk, photobleaching, phototoxicity, technical complexity. challenges arise from conflicting requirements using different fluorophores for labeling with specific properties We propose novel called phasor-based fluorescence spatiotemporal modulation (Phasor-FSTM). This uses time-resolved detection acquire data encoded photons, employs phasor analysis simultaneously separate multiple components, applies create images. Phasor-FSTM enables identification structural components greater spatial accuracy on an enhanced laser scanning confocal microscope single-wavelength laser. To demonstrate capabilities Phasor-FSTM, we performed two-color four-color at resolution ~λ/5 observed interactions organelles live cells during continuous duration over 20 min. Our stands out its simplicity adaptability, seamlessly fitting existing microscopes without requiring lines excitation, which also provides new avenue other technologies based principles build multi-color systems requirement lower budget.

Language: Английский

Primary cilia as dynamic and diverse signalling hubs in development and disease DOI
Pleasantine Mill, Søren T. Christensen, Lotte B. Pedersen

et al.

Nature Reviews Genetics, Journal Year: 2023, Volume and Issue: 24(7), P. 421 - 441

Published: April 18, 2023

Language: Английский

Citations

194
Ångström-resolution fluorescence microscopy DOI Creative Commons
S. Reinhardt, Luciano A. Masullo, Isabelle Baudrexel

et al.

Nature, Journal Year: 2023, Volume and Issue: 617(7962), P. 711 - 716

Published: May 24, 2023

Fluorescence microscopy, with its molecular specificity, is one of the major characterization methods used in life sciences to understand complex biological systems. Super-resolution approaches

Language: Английский

Citations

177
Super-Resolution Microscopy for Structural Cell Biology DOI
Sheng Liu, Philipp Hoess, Jonas Ries

et al.

Annual Review of Biophysics, Journal Year: 2022, Volume and Issue: 51(1), P. 301 - 326

Published: Feb. 4, 2022

Super-resolution microscopy techniques, and specifically single-molecule localization (SMLM), are approaching nanometer resolution inside cells thus have great potential to complement structural biology techniques such as electron for cell biology. In this review, we introduce the different flavors of super-resolution microscopy, with a special emphasis on SMLM MINFLUX (minimal photon flux). We summarize recent technical developments that pushed these localization-based scales review experimental conditions key obtaining data highest quality. Furthermore, give an overview analysis methods highlight studies used gain insights into biologically relevant molecular machines. Ultimately, our perspective what is needed push even further apply them investigating dynamic rearrangements in living cells.

Language: Английский

Citations

149
Endosomal escape: A bottleneck for LNP-mediated therapeutics DOI Creative Commons
Sushmita Chatterjee, Edo Kon, Preeti Sharma

et al.

Proceedings of the National Academy of Sciences, Journal Year: 2024, Volume and Issue: 121(11)

Published: March 4, 2024

Lipid nanoparticles (LNPs) have recently emerged as a powerful and versatile clinically approved platform for nucleic acid delivery, specifically mRNA vaccines. A major bottleneck in the field is release of mRNA-LNPs from endosomal pathways into cytosol cells where they can execute their encoded functions. The data regarding mechanism these escape processes are limited contradicting. Despite extensive research, there no consensus compartment escape, cause inefficient currently lacking robust method to detect escape. Here, we review known mechanisms available methods study this process. We critically discuss limitations challenges possibilities overcome challenges. propose that development robust, quantitative high-throughput techniques timely essential. better understanding process will enable RNA-LNP designs with improved efficiency unlock new therapeutic modalities.

Language: Английский

Citations

139
Stimuli-Responsive Polymer-Based Nanosystems for Cancer Theranostics DOI
Dengshuai Wei, Yong Sun,

Hu Zhu

et al.

ACS Nano, Journal Year: 2023, Volume and Issue: 17(23), P. 23223 - 23261

Published: Dec. 2, 2023

Stimuli-responsive polymers can respond to internal stimuli, such as reactive oxygen species (ROS), glutathione (GSH), and pH, biological enzymes, external lasers ultrasound, etc., by changing their hydrophobicity/hydrophilicity, degradability, ionizability, thus have been widely used in biomedical applications. Due the characteristics of tumor microenvironment (TME), stimuli-responsive that cater specifically TME extensively prepare smart nanovehicles for targeted delivery therapeutic diagnostic agents tissues. Compared conventional drug nanosystems, TME-responsive nanosystems many advantages, high sensitivity, broad applicability among different tumors, functional versatility, improved biosafety. In recent years, a great deal research has devoted engineering efficient polymeric significant improvement made both cancer diagnosis therapy. this review, we summarize some advances involving use polymer nanocarriers delivery, imaging, therapy, theranostics. Various chemical stimuli will be described context nanosystems. Accordingly, groups responsible responsiveness strategies incorporate these into discussed detail. With on topic expending at fast pace, innovative concepts, sequential cascade release, NIR-II multifunctional formulations, emerged popular enhanced performance, which also included here with up-to-date illustrations. We hope review offer valuable insights selection optimization help accelerate future applications treatment.

Language: Английский

Citations

127
Small Size, Big Impact: Recent Progress in Bottom‐Up Synthesized Nanographenes for Optoelectronic and Energy Applications DOI
Zhaoyang Liu, Shuai Fu, Xiaomin Liu

et al.

Advanced Science, Journal Year: 2022, Volume and Issue: 9(19)

Published: Feb. 26, 2022

Bottom-up synthesized graphene nanostructures, including 0D quantum dots and 1D nanoribbons, have recently emerged as promising candidates for efficient, green optoelectronic, energy storage applications. The versatility in their molecular structures offers a large novel library of nanographenes with excellent adjustable optical, electronic, catalytic properties. In this minireview, recent progress on the fundamental understanding properties different state-of-the-art applications optoelectronics are summarized. pristine nanographenes, high emissivity intriguing blinking effect dots, superior charge transport edge-specific electrochemistry various highlighted. Furthermore, it is shown that emerging nanographene-2D material-based van der Waals heterostructures provide an exciting opportunity efficient tunable characteristics. Finally, challenges opportunities field highlighted by offering guidelines future combined efforts synthesis, assembly, spectroscopic, electrical studies well (nano)fabrication to boost toward advanced device

Language: Английский

Citations

112
Real-time denoising enables high-sensitivity fluorescence time-lapse imaging beyond the shot-noise limit DOI Creative Commons
Xinyang Li, Yixin Li, Yiliang Zhou

et al.

Nature Biotechnology, Journal Year: 2022, Volume and Issue: 41(2), P. 282 - 292

Published: Sept. 26, 2022

Abstract A fundamental challenge in fluorescence microscopy is the photon shot noise arising from inevitable stochasticity of detection. Noise increases measurement uncertainty and limits imaging resolution, speed sensitivity. To achieve high-sensitivity beyond shot-noise limit, we present DeepCAD-RT, a self-supervised deep learning method for real-time suppression. Based on our previous framework DeepCAD, reduced number network parameters by 94%, memory consumption 27-fold processing time factor 20, allowing two-photon microscope. high signal-to-noise ratio can be acquired with tenfold fewer photons than standard approaches. We demonstrate utility DeepCAD-RT series photon-limited experiments, including vivo calcium mice, zebrafish larva fruit flies, recording three-dimensional (3D) migration neutrophils after acute brain injury 3D dynamics cortical ATP release. will facilitate morphological functional interrogation biological minimal budget.

Language: Английский

Citations

110
The New Era of High-Throughput Nanoelectrochemistry DOI Creative Commons
Xiangdong Xu, Dimitrios Valavanis,

Paolo Ciocci

et al.

Analytical Chemistry, Journal Year: 2023, Volume and Issue: 95(1), P. 319 - 356

Published: Jan. 10, 2023

ADVERTISEMENT RETURN TO ISSUEPREVReviewNEXTThe New Era of High-Throughput NanoelectrochemistryXiangdong XuXiangdong XuDepartment Chemistry, University Warwick, Coventry CV4 7AL, U.K.More by Xiangdong XuView Biographyhttps://orcid.org/0000-0002-1120-4661, Dimitrios ValavanisDimitrios ValavanisDepartment ValavanisView Biographyhttps://orcid.org/0000-0002-8777-664X, Paolo CiocciPaolo CiocciUniversité Paris Cité, ITODYS, CNRS, F-75013 Paris, FranceMore CiocciView Biography, Samuel ConfederatSamuel ConfederatSchool Electronic and Electrical Engineering Pollard Institute, Leeds, Leeds LS2 9JT, U.K.Bragg Centre for Materials Research, ConfederatView Fabio MarcuccioFabio MarcuccioSchool U.K.Faculty Medicine, Imperial College London, London SW7 2AZ, United KingdomMore MarcuccioView Biographyhttps://orcid.org/0000-0003-4816-2896, Jean-François LemineurJean-François LemineurUniversité LemineurView Actis*Paolo ActisSchool U.K.*[email protected]More ActisView Biographyhttps://orcid.org/0000-0002-7146-1854, Frédéric Kanoufi*Frédéric KanoufiUniversité France*[email KanoufiView Biographyhttps://orcid.org/0000-0002-9784-2380, Patrick R. Unwin*Patrick UnwinDepartment UnwinView Biographyhttps://orcid.org/0000-0003-3106-2178Cite this: Anal. Chem. 2023, 95, 1, 319–356Publication Date (Web):January 10, 2023Publication History Received15 November 2022Published online10 January 2023Published inissue 10 2023https://doi.org/10.1021/acs.analchem.2c05105Copyright © 2023 The Authors. Published American Chemical SocietyRIGHTS & PERMISSIONSACS AuthorChoiceCC: Creative CommonsBY: Credit must be given to the creatorArticle Views1582Altmetric-Citations1LEARN ABOUT THESE METRICSArticle Views are COUNTER-compliant sum full text article downloads since 2008 (both PDF HTML) across all institutions individuals. These metrics regularly updated reflect usage leading up last few days.Citations number other articles citing this article, calculated Crossref daily. Find more information about citation counts.The Altmetric Attention Score is a quantitative measure attention that research has received online. Clicking on donut icon will load page at altmetric.com with additional details score social media presence article. how calculated. Share Add toView InAdd Full Text ReferenceAdd Description ExportRISCitationCitation abstractCitation referencesMore Options onFacebookTwitterWechatLinked InReddit (5 MB) Get e-AlertsSUBJECTS:Ions,Nanoparticles,Nanopores,Probes,Surface charge e-Alerts

Language: Английский

Citations

94
Technological advances in super-resolution microscopy to study cellular processes DOI Creative Commons
Charles Bond, Adriana N. Santiago-Ruiz, Qing Tang

et al.

Molecular Cell, Journal Year: 2022, Volume and Issue: 82(2), P. 315 - 332

Published: Jan. 1, 2022

Language: Английский

Citations

82
4polar-STORM polarized super-resolution imaging of actin filament organization in cells DOI Creative Commons
Caio Vaz Rimoli, César Augusto Valades-Cruz,

Valentina Curcio

et al.

Nature Communications, Journal Year: 2022, Volume and Issue: 13(1)

Published: Jan. 13, 2022

Abstract Single-molecule localization microscopy provides insights into the nanometer-scale spatial organization of proteins in cells, however it does not provide information on their conformation and orientation, which are key functional signatures. Detecting single molecules’ orientation addition to cells is still a challenging task, particular dense cell samples. Here, we present polarization-splitting scheme combines Stochastic Optical Reconstruction Microscopy (STORM) with molecule 2D wobbling measurements, without requiring strong deformation imaged point spread function. This method called 4polar-STORM allows, thanks control its detection numerical aperture, determine both infer 3D orientation. compatible relatively high densities diffraction-limited spots an image, thus ideally placed for investigation protein assemblies cells. We demonstrate potential this actin filament organizations driving adhesion motility.

Language: Английский

Citations

73