Rational Design of Polymethine Dyes with NIR‐II Emission and High Photothermal Conversion Efficiency for Multimodal‐Imaging‐Guided Photo‐Immunotherapy

Advanced Materials, Journal Year: 2023, Volume and Issue: 35(12)

Published: Jan. 11, 2023

Phototheranostics have emerged and flourished as a promising pattern for cancer theranostics owing to their precise photoinduced diagnosis therapeutic meet the demands of precision medicine. The information effect are directly determined by fluorescence imaging ability photothermal conversion efficiency (PCE) phototheranostic agents. Hence, how balance competitive radiative nonradiative processes agents is key factor evaluate effect. Herein, molecules named ICRs with high photostaibility rationally designed, exhibiting emission in second near-infrared window (NIR-II, 1000-1700 nm) PCE, which related strong donor-acceptor (D-A) interaction reorganization energy Noteworthily, ICR-Qu stronger D-A large-sized conjugated unit encapsulated nanoparticles exhibits PCE (81.1%). In addition, ICR-QuNPs used (FLI), photoacoustic (PAI), (PTI) guide deep-tissue photonic hyperthermia, achieving removal inhibition breast cancer. Furthermore, combined α-PD-1, show huge potential be facile efficient tool photo-immunotherapy. More importantly, this study not only reports an "all-in-one" polymethine-based agent, but also sheds light on exploration versatile organic future practical applications.

Language: Английский

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Minimal genetically encoded tags for fluorescent protein labeling in living neurons

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

Published: Jan. 14, 2022

Abstract Modern light microscopy, including super-resolution techniques, has brought about a demand for small labeling tags that bring the fluorophore closer to target. This challenge can be addressed by unnatural amino acids (UAAs) with bioorthogonal click chemistry. The minimal size of UAA and possibility couple fluorophores directly protein interest single-residue precision in living cells make unique. Here, we establish primary neurons use it fixed-cell, live-cell, dual-color pulse–chase, microscopy neurofilament chain (NFL). We also show combined CRISPR/Cas9 genome engineering tagging endogenous NFL. Due its versatile nature compatibility advanced multicolor anticipate will contribute novel discoveries neurobiology field.

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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Highly catalytic and durable nanocellulose fibers-based nanoporous membrane film for efficient organic pollutant degradation

Journal of Water Process Engineering, Journal Year: 2023, Volume and Issue: 53, P. 103620 - 103620

Published: March 15, 2023

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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Ultra-photostable small-molecule dyes facilitate near-infrared biophotonics

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: March 22, 2024

Abstract Long-wavelength, near-infrared small-molecule dyes are attractive in biophotonics. Conventionally, they rely on expanded aromatic structures for redshift, which comes at the cost of application performance such as photostability, cell permeability, and functionality. Here, we report a ground-state antiaromatic strategy showcase concise synthesis 14 cationic aminofluorene with mini (molecular weights: 299–504 Da) distinct spectra covering 700–1600 nm. Aminofluorene cell-permeable achieve rapid renal clearance via simple 44 Da carboxylation. This accelerates optical diagnostics injury by 50 min compared to existing macromolecular approaches. We develop compact molecular sensing platform vivo intracellular sensing, demonstrate versatile applications these multispectral fluorescence optoacoustic imaging. find that aromaticity reversal upon electronic excitation, indicated magnetic descriptors, not only reduces energy bandgap but also induces strong vibronic coupling, resulting ultrafast excited-state dynamics unparalleled photostability. These results support argument antiaromaticity useful design rule dye development, enabling performances essential modern

Language: Английский

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Advancing cell biology with nanoscale fluorescence imaging: essential practical considerations

Trends in Cell Biology, Journal Year: 2024, Volume and Issue: 34(8), P. 671 - 684

Published: Jan. 5, 2024

Here, we provide an overview of the key factors to consider when using fluorescence nanoscopy (FN), including biological questions that can be addressed and aspects might improve reliability effectiveness FN experiments.We cover main related sample preparation, selection appropriate fixation, affinity-based labels, fluorescent dyes.We discuss current limitations possible future developments in field would facilitate a broader application FN.We multiplexing possibilities (allowing simultaneous detection multiple targets single experiment), live cell imaging for study cellular molecular dynamic processes, quantitative workflows. Recently, biologists have gained access several far-field (FN) technologies allow observation components with ~20 nm resolution. is revolutionizing biology by enabling visualization previously inaccessible subcellular details. While technological advances microscopy are critical field, optimal preparation labeling equally important often overlooked experiments. In this review, methodological experimental must considered performing FN. We present concepts dyes, multiplexing, approaches, microscopy. Consideration these greatly enhances FN, making it exquisite tool numerous applications. Conventional microscopy, widefield confocal has been essential studying morphology composition various organelles localization molecules. The resolution (see Glossary) techniques, usually above ~200 nm, limit macromolecular arrangements nanoscale structures. processes (<20 nm) major step forward because bridges world macromolecules. This effort facilitated recent decades use electron (EM), which provides morphological information at level [1.Winey M. et al.Conventional transmission microscopy.Mol. Biol. Cell. 2014; 25: 319-323Crossref PubMed Google Scholar] expenses limited identifications lack applicability. These overcome achieved resolving capabilities progressively reaching those attained EM. Several approaches based on enable researchers address <20 were difficult answer classical Paradigmatic examples applications include characterization periodicity actin rings synaptic sites neurons, structure nuclear pore complexes, maturation viral particles, organization mitochondrial cristae, mechanisms apoptosis, functioning signaling pathways [2.Fornasiero E.F. Opazo F. Super-resolution biologists.BioEssays. 2015; 37: 436-451Crossref Scholar]. Recent studies reached extremely high resolutions [3.Reinhardt S.C.M. al.Ångström-resolution microscopy.Nature. 2023; 617: 711-716Crossref Scopus (21) even enabled follow events such as stepping behavior kinesin vitro cells [4.Wolff J.O. al.MINFLUX dissects unimpeded walking kinesin-1.Science. 379: 1004-1010Crossref (20) Scholar,5.Deguchi T. al.Direct motor protein living MINFLUX.Science. 1010-1015Crossref (22) render individual molecules, GABA receptors, detail almost comparable cryo-EM [6.Shaib A.H. al.Visualizing proteins expansion microscopy.bioRxiv. (Published online March 10, 2023. https://doi.org/10.1101/2022.08.03.502284)Google compendium summarizes practical keep mind harnessing power nanoscopic biology. By bridging theory practice, roadmap researchers, equipping them know-how successfully navigate intricacies implementing, executing, deriving meaningful data from design experiments starts most suitable technique, each its own specific strengths (Figure 1A ; reviews different see [7.Jacquemet G. al.The biologist's guide super-resolution microscopy.J. Cell Sci. 2020; 133jcs240713Crossref (32) Scholar,8.Bond C. al.Technological processes.Mol. 2022; 82: 315-332Abstract Full Text PDF (25) Scholar]). Two strategies currently able reliably <20-nm samples: camera-based single-molecule (SMLM) [9.Rust M.J. al.Sub-diffraction-limit stochastic optical reconstruction (STORM).Nat. Methods. 2006; 3: 793-796Crossref (6072) minimal photon fluxes (MINFLUX) [10.Balzarotti al.Nanometer tracking molecules fluxes.Science. 2017; 355: 606-612Crossref (657) Based utilized perform on–off switching fluorophores required obtain super-resolved image, SMLM names; example, (STORM), photoactivated (PALM), DNA-points accumulation topography (DNA-PAINT) [11.Jungmann R. al.Single-molecule kinetics transient binding DNA origami.Nano Lett. 2010; 10: 4756-4761Crossref (576) addition two strategies, depending exact settings, stimulated emission depletion (STED) [12.Hell S.W. Wichmann J. Breaking diffraction emission: stimulated-emission-depletion microscopy.Opt. 1994; 19: 780Crossref achieve ~50 below, although laser not compatible conventional samples. technologies, approach aimed enlarging sample, then imaged either diffraction-limited or [13.Chen al.Optical imaging. Expansion microscopy.Science. 347: 543-548Crossref (858) With exception microcopy, specimens, opening avenue understanding dynamics native conditions (Box 1). Overall, four potential uncover as-yet unexplored detail. At same time, their capability requires protocols introduction artifacts tools linkage errors.Box 1Nanometer-scale cells: there yet?FN visualizes entire structures over time 'whole', allowing changes, recommended endoplasmic reticulum, chemical fixation introduces visible [19.Hoffman D.P. al.Correlative three-dimensional block-face whole vitreously frozen cells.Science. 367eaaz5357Crossref (33) our opinion, least connected implementation detailed herein.PhototoxicityPhototoxicity arises absorbed light generates free radicals reactive oxygen species, ultimately causing genomic damage, stress, degradation Figure 1D text). Light both endogenous additionally increase local temperature sample. To some effects, combinations image adaptive illumination [75.Heine al.Adaptive-illumination STED nanoscopy.Proc. Natl. Acad. U. S. A. 114: 9797-9802Crossref (0) Scholar], event-triggered emerging [76.Alvelid al.Event-triggered imaging.Nat. 1268-1275Crossref (15) Furthermore, novel fluorophore classes self-blinking dyes red photoswitchable exist, do require blue excited lower doses [77.Pennacchietti al.Fast reversibly photoswitching live-cell RESOLFT nanoscopy.Nat. 2018; 15: 601-604Crossref (58) Scholar].Low acquisition frequencyLow frequency resolution, but compromises interpretation fast processes. SMLM, limiting factor sufficient number while, scanning-based steps brightness size view. Reducing view spent pixel (dwell time) speed up imaging, decreased contextual SNR. will strongly benefit parallelization deep learning temporal performances.Availability impact cellsThe availability cannot ignored. Some probes drugs bind target affinity, interfering physiology targeted molecule (e.g., phalloidin). An alternative genetically encoded tags or, better, combined genome-editing [78.Bottanelli al.A physiological role ARF1 formation bidirectional tubules Golgi.Mol. 28: 1676-1687Crossref Table 1 text).Imaging depth large viewImaging many samples monolayers. Although feasibility specialized demonstrated [79.Kim al.Oblique-plane tissues small intact animals.Nat. 2019; 16: 853-857Crossref (54) deeper than 10–50 μm methods challenging. engineered illumination, optics, restoration algorithms, multiphoton excitation.Ultimately, best technology selected setup precise question being addressed, controls phototoxic cause damage considered. should done settings less perturbation troubleshooting conditions. For far-red commonly used compared 405-laser irradiance. herein. Phototoxicity Low performances. Availability Imaging excitation. Ultimately, short 'no', all problems solved. Careful consideration given whether necessary questions. determine interest (POI) localized lysosomes mitochondria, POI outer membrane inner matrix, apart. general, strength increased precision biomolecules 1B,C). A simple rule thumb decide needed understand spatial order tens nanometers allows formulation fundamentally hypotheses process under investigation. serendipitous observations made and, cases, revealed new observable [14.Xu K. al.Actin, spectrin, associated form periodic cytoskeletal axons.Science. 2013; 339: 452-456Crossref (888) reason, exploratory Very high-resolution informative, could ignored become challenges Live ultimate goal studies, still due phototoxicity early stage foreseeable relevant near future. fixed already changing field; therefore, concentrate here specimens 1D–F). workflow 'non-live' first optimized avoid artifacts. After antibodies, reveal identities position reporter proximity. relative absolute biologically numbers images (quantitative FN), further measures considerations taken. Box 2, review issues explain why density, stoichiometry, error context. Finally, 3, conditions.Box 2Quantitative FN: versus observed moleculesQuantitative detected matches (or, more precisely, correlates) real Therefore, prerequisites controlled efficiency ability detect fluorophores.A monovalent affinity carrying (or of) reporting 2A Moreover, utilize covalently linked ensure stably labeled preferred. multivalent polyclonal reagents stochastically secondary antibodies approximately one six fluorophores) avoided since correlation between reporters inconsistent.Even ideal case decorated label, possibility exists only decorating label functional. Indeed, inactivated, damaged, during procedure 1E Detecting densely challenging, problem known crowding. When distances single-digit nanometer scale, photophysical interactions occur, resulting undesired alterations properties. happens structure, Förster resonance energy transfer (FRET) H-dimer if separated molecular-scale [80.Ogawa al.H-Type dimer fluorophores: mechanism activatable, vivo imaging.ACS Chem. 2009; 4: 535-546Crossref (153) Notably, reported antibody [81.Helmerich D.A. al.Multiple-labeled behave like emitters buffer.ACS Nano. 14: 12629-12641Crossref (13) serving 'super emitter' while others remain dark state. crowding reversed, physically creating distance Other DNA-PAINT, deal differently modulating concentration probe light-controllable [82.Raymo F.M. Photoactivatable synthetic nanoscale.J. Phys. 2012; 2379-2385Crossref (60) 2D text).While quantification easily achieved, molecule-counting proposed comprehensive recently published challenge [83.Hugelier al.Quantitative microscopy.Annu. Rev. Biophys. 52: 139-160Crossref (1) Importantly, need calibration benchmarked against markers biochemically western blot, liquid chromatography, mass spectrometry).Box 3Fluorophore no one-size-fits-all solutionsA variety developed fulfill requirement technique (Table I). Factors selecting charge, quantum yield, photostability 2C photostable fluorophores, MINFLUX rely switch non-emitting emitting states. mechanisms, refer reader text.The scaffolds cyanines rhodamines. Among cyanines, Alexa Fluor 647 gold standard blinks presence reducing agents UV [84.Berlier J.E. comparison long-wavelength Cy Dyes: bioconjugates.J. Histochem. Cytochem. 2003; 51: 1699-1712Crossref (229) Rhodamines relatively modified tune spectral properties [85.Grimm J.B. general method optimize functionalize red-shifted rhodamine dyes.Nat. 17: 815-821Crossref (85) permeability [86.Lukinavičius near-infrared proteins.Nat. 5: 132-139Crossref (648) equilibrium open closed non-fluorescent forms. regulation latter induces cell-compatible spontaneous blinking Scholar].Other frequently coumarin, oxazine, BODIPY 2E Coumarins among smallest generate variants Stokes shift, advantageous low background [87.Nizamov al.Phosphorylated 3-heteroarylcoumarins nanoscopy.Chem. Eur. 18: 16339-16348Crossref (45) Oxazines [88.Wombacher al.Live-cell trimethoprim conjugates.Nat. 7: 717-719Crossref (282) absorbance, extinction coefficient, 'blink' buffers containing oxidizing agents. valued sharp absorption spectra very yield coefficient [89.Kowada al.BODIPY-based cells.Chem. Soc. 44: 4953-4972Crossref highly hydrophobic nature poor off-switching properties, light-dose photoactivatable make attractive [90.Wijesooriya C.S. localization-based imaging.Angew. Int. Ed. 57 (12685–1268)Crossref (74) Scholar].In after identified, always considering specifications available instrument lasers detectors). choice also driven needs, multicolor autofluorescence reduce background, counting naked may themselves affinities lipophilic stain membranes). evaluated designing experiments, (reporters) without targeting moiety whenever 2B recommend inexperienced users consult expert select dye application.Table IProperties dyesFluorophore classCoumarinsRhodaminesCyaninesBODIPYsOxazinesCommercial examplesAlexa 350, Pacific BlueAlexa 488, silicon-rhodamine, TMRAlexa 647, 555, Cy5BODIPY FL, TMRAtto 655, Atto 680Spectral range (nm)360–700500–750500–1000500–700600–750Extinction (cm–1M–1)15 000–60 00080 000–150 000130 000–250 00060 000–100 000110 000–130 000Quantum yield0.4–0.90.1–0.90.1–0.60.8–0.90.1–0.6Photostability+++++++++++Compatibility methodsSTED, SMLMSTED, MINFLUX, microscopySTED, SMLMSMLM Open table tab Quantitative fluorophores. inconsistent. Even sin

Language: Английский

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Fluorescent chemosensors facilitate the visualization of plant health and their living environment in sustainable agriculture

Chemical Society Reviews, Journal Year: 2024, Volume and Issue: 53(13), P. 6992 - 7090

Published: Jan. 1, 2024

Globally, 91% of plant production encounters diverse environmental stresses. Fluorescent chemosensors are effective for monitoring health and environment that promotes the development sustainable agriculture.

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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