Photoactivatable Plasma Membrane Probe Through Self-Triggered Photooxidation Cascade for Live Super-Resolution Microscopy DOI Creative Commons
Sonia Pfister, Valentine Le Berruyer, Kyong Tkhe Fam

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: May 30, 2024

Abstract Super-resolution imaging based on the localization of single emitters requires a spatio-temporal control ON and OFF state. To this end, photoactivatable fluorophores are adapted as they can be turned upon light irradiation. Here we present concept called Self-Triggered Photooxidation Cascade (STPC) photooxidation plasma membrane targeted leuco-rhodamine (LRhod-PM), non-fluorescent reduced form rhodamine probe. Upon visible irradiation small number oxidized rhodamines, Rhod-PM, acts photosensitizer to generate singlet oxygen capable oxidize state LRhod-PM. We showed that phenomenon is kinetically favored by high local concentration propagates quickly when probe embedded in bilayers. In addition, close proximity dyes favors photobleaching. At single-molecule level, concomitant activation/bleaching phenomena allow reaching blinking regime enabling microscopy for super-resolution live cellular membranes.

Language: Английский

Hybrid Small-Molecule/Protein Fluorescent Probes DOI
Masafumi Minoshima, Shahi Imam Reja,

Ryu Hashimoto

et al.

Chemical Reviews, Journal Year: 2024, Volume and Issue: 124(10), P. 6198 - 6270

Published: May 8, 2024

Hybrid small-molecule/protein fluorescent probes are powerful tools for visualizing protein localization and function in living cells. These hybrid constructed by diverse site-specific chemical labeling approaches through reactions to exogenous peptide/small tags, enzymatic post-translational modifications, bioorthogonal genetically incorporated unnatural amino acids, ligand-directed reactions. The employed imaging trafficking, conformational changes, bioanalytes surrounding proteins. In addition, facilitate visualization of dynamics at the single-molecule level defined structure with super-resolution imaging. this review, we discuss development bioimaging applications based on hybrids.

Language: Английский

Citations

24
Photoactivation Inducing Multifunctional Coupling of Fluorophore for Efficient Tumor Therapy In Situ DOI
Weijie Zhang,

Yunxia Lv,

Fangjun Huo

et al.

Advanced Materials, Journal Year: 2024, Volume and Issue: 36(23)

Published: Feb. 15, 2024

Abstract Photoactivatable molecules, with high‐precision spatialtemporal control, have largely promoted bioimaging and phototherapy applications of fluorescent dyes. Here, the first photoactivatable sensor ( BI ) is described that can be triggered by broad excitation light (405–660 nm), which further undergoes intersystem crossing H‐atom transfer processes to forming superoxide anion radicals (O 2 −• carbon radicals. Particularly, photoinduced gain carbon‐centered BI• allows for radical‐radical coupling afford combined crosslink product BI─BI ), would oxidized in presence O produce an extended conjugate system near infrared emission (820 nm). Besides, photochemically generated Cy─BI possesses ultra‐high photothermal conversion efficiency up 90.9%, optimized potential. What′s more, Western Blot assay reveals both photoproduct efficiently inhibit expression CHK1, irradiation induces apoptosis ultimately enhances phototherapeutic effects. Thus, combination cell cycle block inducing apoptosis, photodynamic therapy treatments significantly suppress solid tumor vivo antitumor efficacy explorations. This a novel finding developing as well applicability photoimaging tumor‐related areas.

Language: Английский

Citations

19
Biosynthesis and genetic encoding of activated nitriles for fast protein conjugation and tunable fluorogenic labeling DOI
Elwy H. Abdelkader, Haocheng Qianzhu, Gottfried Otting

et al.

Chem, Journal Year: 2025, Volume and Issue: unknown, P. 102385 - 102385

Published: Jan. 1, 2025

Language: Английский

Citations

2
HaloTag‐Based Reporters for Fluorescence Imaging and Biosensing DOI Creative Commons
Alexander Cook, Franziska Walterspiel, Claire Deo

et al.

ChemBioChem, Journal Year: 2023, Volume and Issue: 24(12)

Published: Feb. 23, 2023

Abstract Visualizing the structure and dynamics of biomolecules is critical to understand biological function, requires methods fluorescently label targets interest in their cellular context. Self‐labelling proteins, which combine a genetically encoded tag with small‐molecule fluorophore, have attracted considerable attention for this purpose, as they can overcome limitations fluorescent proteins. Among them, HaloTag protein most broadly used, showing fast specific labelling small, easy functionalize cell‐permeant ligand. Synthetic chemistry engineering provided portfolio powerful imaging tools exploiting HaloTag, along general optimize adapt them applications. Here, we provide an overview reporters based on biosensing, highlighting strategies

Language: Английский

Citations

36
Targeted Photoconvertible BODIPYs Based on Directed Photooxidation-Induced Conversion for Applications in Photoconversion and Live Super-Resolution Imaging DOI
Lazare Saladin, Victor Breton, Valentine Le Berruyer

et al.

Journal of the American Chemical Society, Journal Year: 2024, Volume and Issue: 146(25), P. 17456 - 17473

Published: June 11, 2024

Photomodulable fluorescent probes are drawing increasing attention due to their applications in advanced bioimaging. Whereas photoconvertible can be advantageously used tracking, photoswitchable constitute key tools for single-molecule localization microscopy perform super-resolution imaging. Herein, we shed light on a red and far-red BODIPY, namely, BDP-576 BDP-650, which possess both properties of conversion switching. Our study demonstrates that these pyrrolyl-BODIPYs convert into typical green- red-emitting BODIPYs perfectly adapted microscopy. We also showed this undergo Directed Photooxidation Induced Conversion, photoconversion mechanism recently introduced, where the pyrrole moiety plays central role. These unique features were develop targeted toward different organelles or subcellular units (plasma membrane, mitochondria, nucleus, actin, Golgi apparatus, etc.) using chemical targeting moieties Halo tag. notably BDP-650 could track intracellular vesicles over more than 20 min two-color imagings with laser scanning confocal microscopy, demonstrating its robustness. The switching photoconverters studied at level then successfully live epithelial cells neurons. Both membrane- mitochondria- decipher membrane 3D architecture mitochondrial dynamics nanoscale. This builds bridge between photoswitching undergoing directed photooxidation shows versatility efficacy

Language: Английский

Citations

11
Organic Fluorophores with Large Stokes Shift for Bioimaging and Biosensing DOI
Hong Liu, Gangwei Jiang, Guoliang Ke

et al.

ChemPhotoChem, Journal Year: 2023, Volume and Issue: 8(5)

Published: Dec. 14, 2023

Abstract Fluorophores and fluorophores‐based probes or labels can visualize the structures content fluctuations of biomolecules, have made great progress in broad range biomedicine fields. However, many commercially available fluorophores suffer from small Stokes shift, which results insufficient signal‐to‐noise ratio self‐quenching advanced imaging techniques. Moreover, shift also hinders application some complex imaging, such as single‐excitation multicolor imaging. In past two decades, effects been to enlarge fluorophores. this review, we clarified reasons for building on structural analysis fluorophores, systematically summarize methods modification increase present representative applications with large sensing.

Language: Английский

Citations

15
Bioorthogonal Caging-Group-Free Photoactivatable Probes for Minimal-Linkage-Error Nanoscopy DOI Creative Commons
Ayse Aktalay, Richard Lincoln,

Lukas Heynck

et al.

ACS Central Science, Journal Year: 2023, Volume and Issue: 9(8), P. 1581 - 1590

Published: July 26, 2023

Here we describe highly compact, click compatible, and photoactivatable dyes for super-resolution fluorescence microscopy (nanoscopy). By combining the xanthone (PaX) core with a tetrazine group, achieve minimally sized sensitive molecular dyads selective labeling of unnatural amino acids introduced by genetic code expansion. We exploit excited state quenching properties group to attenuate photoactivation rates PaX, further reduce overall emission photogenerated fluorophore, providing two mechanisms selectivity off-target signal. Coupled MINFLUX nanoscopy, employ our in minimal-linkage-error imaging vimentin filaments, demonstrating molecular-scale precision fluorophore positioning.

Language: Английский

Citations

13
PyrAtes: Modular Organic Salts with Large Stokes Shifts for Fluo‐rescence Microscopy DOI Creative Commons
Iakovos Saridakis, Margaux Riomet, Oliver John V. Belleza

et al.

Angewandte Chemie International Edition, Journal Year: 2024, Volume and Issue: 63(19)

Published: April 3, 2024

Abstract The deployment of small‐molecule fluorescent agents plays an ever‐growing role in medicine and drug development. Herein, we complement the portfolio powerful fluorophores, reporting serendipitous discovery development a novel class with imidazo[1,2‐a]pyridinium triflate core, which term PyrAtes . These fluorophores are synthesized single step from readily available materials (>60 examples) display Stokes shifts as large 240 nm, while also reaching NIR−I emissions at λ max long 720 nm. Computational studies allow platform for prediction Em Furthermore, demonstrate compatibility these live cell imaging HEK293 cells, suggesting potent intracellular markers.

Language: Английский

Citations

5
Anion-induced opposite mechanochromic and thermochromic emission directions of protonated hydrazones DOI
Cheng Qian,

Lixia Xie,

Lijie Liu

et al.

Science China Materials, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 4, 2024

Language: Английский

Citations

5
Toolbox of Clickable Benzylguanines for Labeling of HoxB8-Derived Macrophages via SNAP-Tag and Bioorthogonal Chemistry DOI

Jonas Schöning,

Lukas Rösner,

Dominic Alexej Depke

et al.

Bioconjugate Chemistry, Journal Year: 2025, Volume and Issue: 36(1), P. 34 - 43

Published: Jan. 6, 2025

Inflammation is a dynamic process which importantly involves migration of immune cells. Understanding the onset, acute phase and resolution inflammation greatly facilitated by their in vivo imaging. However, cells are sensitive, difficult to genetically manipulate prone changes response contact, hindering application well-established cell labeling methods. We generated first reported SNAP-tag expressing conditionally immortalized early hematopoietic progenitor (ER-HoxB8) synthesized benzylguanine (BG) analogs with bioorthogonal groups for mediated surface labeling. Comparative investigation positive HeLa cells, ER-HoxB8 ER-HoxB8-derived macrophages as well neutrophiles revealed remarkable differences depending on group fluorescent reporter used. were efficiently labeled BG bearing an azide dioxolan-fused trans-cyclooctene (dTCO). When we differentiated into macrophages, was less bright due lower expression only tetrazine ligation dTCO proved suitable cell-type specific These results show that exploring different reactions important address challenge Combining click chemistry provides modular approach cell-specific combination presents improved system macrophages.

Language: Английский

Citations

0