Single-Molecule Localization Microscopy and Tracking with a Fluorescent Mechanosensitive Probe DOI

Jimmy Maillard,

Ewa Grassin,

Ekaterina Bestsennaia

et al.

The Journal of Physical Chemistry B, Journal Year: 2024, Volume and Issue: 128(33), P. 7997 - 8006

Published: Aug. 9, 2024

A milestone in optical imaging of mechanical forces cells has been the development family flipper fluorescent probes able to report membrane tension noninvasively living through their fluorescence lifetime. The specifically designed Flipper-CF3 probe with an engineered inherent blinking mechanism was recently introduced for super-resolution microscopy lipid ordered membranes but too dim be detected disordered at single-molecule level (García-Calvo, J. Am. Chem. Soc. 2020, 142(28), 12034–12038). We show here that original and commercially available Flipper-TR is compatible based resolves both liquid giant unilamellar vesicles below diffraction limit. Single molecules were additionally tracked bilayers, enabling distinguish varying composition from diffusion coefficient probe. Differences brightness between originate steady-state absorption properties. general compatibility scaffold detection further shown experiments targetable derivatives.

Language: Английский

Recent Development of Advanced Fluorescent Molecular Probes for Organelle-Targeted Cell Imaging DOI Creative Commons
Sha Lu, Zhi‐Qi Dai, Yunxi Cui

et al.

Biosensors, Journal Year: 2023, Volume and Issue: 13(3), P. 360 - 360

Published: March 8, 2023

Fluorescent molecular probes are very powerful tools that have been generally applied in cell imaging the research fields of biology, pathology, pharmacology, biochemistry, and medical science. In last couple decades, numerous endowed with high specificity to particular organelles designed illustrate intracellular images more detail at subcellular level. Nowadays, development biology has enabled investigation process go deeply into cells, even Therefore, can sketch a organelle’s location while responding certain parameters evaluate bioprocesses under urgent demand. It is significant understand basic ideas organelle properties, as well vital substances related each unique organelle, for design efficiency. this review, we summarize representative multifunctional fluorescent developed decade. We focus on specially target nuclei, mitochondria, endoplasmic reticulums, lysosomes. section, first briefly introduce significance properties different organelles. then discuss how make them highly organelle-specific. Finally, also consider constructed endow additional functions recognize physical/chemical signals targeted Moreover, perspective challenges future applications specific proposed. hope review provide researchers conceptual information about developing imaging, assisting scientists interested biochemistry accelerate their scientific studies.

Language: Английский

Citations

12
Molecular Rotors: Fluorescent Sensors for Microviscosity and Conformation of Biomolecules DOI Creative Commons
Miguel Páez-Pérez, Marina K. Kuimova

Angewandte Chemie, Journal Year: 2023, Volume and Issue: 136(6)

Published: Oct. 19, 2023

Abstract The viscosity and crowding of biological environment are considered vital for the correct cellular function, alterations in these parameters known to underly a number pathologies including diabetes, malaria, cancer neurodegenerative diseases, name few. Over last decades, fluorescent molecular probes termed rotors proved extremely useful exploring viscosity, crowding, underlying interactions biologically relevant settings. In this review, we will discuss basic principles underpinning functionality review advances their use as sensors lipid order, protein conformation, temperature non‐canonical nucleic acid structures live cells other

Language: Английский

Citations

11
Core‐Alkynylated Fluorescent Flippers: Altered Ultrafast Photophysics to Track Thick Membranes DOI Creative Commons

Khurnia Krisna Puji Pamungkas,

Ina Fureraj,

Lea Assies

et al.

Angewandte Chemie International Edition, Journal Year: 2024, Volume and Issue: 63(31)

Published: May 17, 2024

Fluorescent flippers have been introduced as small-molecule probes to image membrane tension in living systems. This study describes the design, synthesis, spectroscopic and imaging properties of that are elongated by one two alkynes inserted between push pull dithienothiophene domains. The resulting mechanophores combine characteristics flippers, reporting on physical compression ground state, molecular rotors, torsional motion excited take their photophysics new level sophistication. Intensity ratios broadened excitation bands from differently twisted conformers core-alkynylated thus report mechanical compression. Lifetime boosts ultrafast excited-state planarization lifetime drops competitive intersystem crossing into triplet states viscosity. In standard lipid bilayer membranes, too long for leaflet tilt or extend disordered interleaflet space, which preserves rotor-like disorder weak, blue-shifted fluorescence. Flipper-like occurs only highly ordered membranes matching thickness, where they light up selectively these thick with red-shifted, sharpened maxima, high intensity lifetime.

Language: Английский

Citations

4
Anionic Cyanine Membrane Probes for Live Cells and In Vivo Fluorescence Imaging DOI
Dmytro I. Danylchuk, Igor Khalin, Yelisetty Venkata Suseela

et al.

Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: April 11, 2025

Molecular probes for cell plasma membranes are indispensable fluorescence imaging. Herein, we present an array of five anionic cyanine-based turn-on membrane with emission spanning from green to near infrared. They analogous the commonly used MemBright probe family, where two zwitterionic anchor groups replaced sulfonates dodecyl chains. The developed provide selective wash-free staining live cells in vitro, featuring improved brightness and slower internalization inside cells. In comparison protein-based (wheat germ agglutinin) markers, new better poorly accessible cell-cell contacts. A key challenge is stain directly vivo. During vivo brain tissue imaging living mice by two-photon microscopy, cyanine allowed us visualize detail pyramidal neurons high image quality, clearly resolving neuron soma, dendrites dendritic spines, axons axonal boutons. constitute important extension toolbox research.

Language: Английский

Citations

0
Elastic remodeling of model and cell membranes by rotating ATP synthase DOI Creative Commons
David Valdivieso González,

Miguel A. Sacristán,

Paolo Natale

et al.

Cell Reports Physical Science, Journal Year: 2025, Volume and Issue: unknown, P. 102567 - 102567

Published: April 1, 2025

Language: Английский

Citations

0
Fluorogenic In Situ Thioacetalization: Expanding the Chemical Space of Fluorescent Probes, Including Unorthodox, Bifurcated, and Mechanosensitive Chalcogen Bonds DOI Creative Commons
Xiaoxiao Chen, Rosa M. Gomila, Juan Manuel GARCIA ARCOS

et al.

JACS Au, Journal Year: 2023, Volume and Issue: 3(9), P. 2557 - 2565

Published: Aug. 21, 2023

Progress with fluorescent flippers, small-molecule probes to image membrane tension in living systems, has been limited by the effort needed synthesize twisted push-pull mechanophore. Here, we move a higher oxidation level introduce new design paradigm that allows screening of flipper rapidly, at best situ. Late-stage clicking thioacetals and acetals simultaneous attachment targeting units interfacers exploration critical chalcogen-bonding donor same time. Initial studies focus on plasma develop chemical space thioacetals, from acyclic amino acids cyclic 1,3-heterocycles covering dioxanes as well dithiolanes, dithianes, dithiepanes, derived also classics biology like cysteine, lipoic acid, asparagusic DTT, epidithiodiketopiperazines. From functional point view, sensitivity imaging cells could be doubled, lifetime differences FLIM images increasing 0.55 1.11 ns. theoretical complexity mechanically coupled chalcogen bonding is explored, revealing, among others, intriguing bifurcated bonds.

Language: Английский

Citations

10
Fluorescence Intensity and Lifetime Imaging of A Viscosity Sensitive Zwitterionic Amphiphile for Cell Membrane Tension and Apoptosis Bodies DOI
Yifan Liu, Bingpeng Guo, Xiaoling Zhang

et al.

Sensors and Actuators B Chemical, Journal Year: 2025, Volume and Issue: unknown, P. 137474 - 137474

Published: Feb. 1, 2025

Language: Английский

Citations

0
Investigation of Symmetric and Nonsymmetric Dithienothiophene-dioxide Derivatives as Fluorescent Emitters for Luminescent Solar Concentrators DOI
Matteo Bartolini, Alberto Picchi,

Hanna Pryshchepa

et al.

ACS Applied Energy Materials, Journal Year: 2025, Volume and Issue: unknown

Published: April 4, 2025

Language: Английский

Citations

0
Recent developments in probing the levels and flux of selected organellar cations as well as organellar mechanosensitivity DOI Creative Commons
Taufiq Rahman, Sandip Patel

Current Opinion in Chemical Biology, Journal Year: 2025, Volume and Issue: 87, P. 102600 - 102600

Published: May 3, 2025

Electrochemical gradients exist not only across the plasma membrane (PM) but also membranes of organelles. Various endomembrane-localised ion channels and transporters have been identified, activity which is critical for organellar (and cellular) ionic homeostasis that underpins diverse cellular processes. Aberrant flux underlies several diseases, identifying as potential drug targets. Therefore, need probing functions these proteins in situ cannot be overemphasised. The acidic interior a few organelles well dynamic nature most historically presented challenges reliable estimation luminal concentrations. But there significant methodological technical advancements by now, allowing measurement levels specific ions within their endomembranes with increasing precision. Evidence continues to amass reporting mechanosensitivity its physiological significance. Here we highlight some recent developments tools techniques measuring movement selected cations mechanosensitivity.

Language: Английский

Citations

0
Advancing Dynamic Polymer Mechanochemistry through Synergetic Conformational Gearing DOI
Nnamdi Ofodum, Qingkai Qi, Richard Chandradat

et al.

Journal of the American Chemical Society, Journal Year: 2024, Volume and Issue: 146(26), P. 17700 - 17711

Published: June 18, 2024

Harnessing mechanical force to modulate material properties and enhance biomechanical functions is essential for advancing smart materials bioengineering. Polymer mechanochemistry provides an emerging toolkit exploring unconventional chemical transformations modulating molecular structures through force. One of the key challenges developing innovative force-sensing mechanisms precise

Language: Английский

Citations

3