Flow Cytometry Analysis of Perturbations in the Bacterial Cell Envelope Enabled by Monitoring Generalized Polarization of the Solvatochromic Peptide UNR-1 DOI

Lucille Weiss,

Dominique Bonnet, Dmytro Dziuba

et al.

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 97(1), P. 622 - 628

Published: Dec. 30, 2024

The worldwide spread of antibiotic resistance is considered to be one the major health threats society. While developing new antibiotics crucial, there also a strong need for next-generation analytical methods studying physiological state live bacteria in heterogeneous populations and their response environmental stress. Here we report single-cell high-throughput method monitor changes bacterial cell envelope stress based on ratiometric flow cytometry. We used sensitive fluorescent molecular probe, Nile Red-based solvatochromic antimicrobial peptide UNR-1, with defined cellular localization Gram-positive Gram-negative bacteria. developed robust protocol calculating generalized polarization (GP) fluorescence adapted Our methodology enabled rapid detection perturbations caused by exposure antibiotics, heat shock, other factors.

Language: Английский

Single-Molecule Spectroscopy and Super-Resolution Mapping of Physicochemical Parameters in Living Cells DOI
Megan A. Steves, Changdong He, Ke Xu

et al.

Annual Review of Physical Chemistry, Journal Year: 2024, Volume and Issue: 75(1), P. 163 - 183

Published: Feb. 16, 2024

By superlocalizing the positions of millions single molecules over many camera frames, a class super-resolution fluorescence microscopy methods known as single-molecule localization (SMLM) has revolutionized how we understand subcellular structures past decade. In this review, highlight emerging studies that transcend outstanding structural (shape) information offered by SMLM to extract and map physicochemical parameters in living mammalian cells at levels. encoding/decoding high-dimensional information-such emission excitation spectra, motion, polarization, lifetime, beyond-for every molecule, mass accumulating these measurements for molecules, such multidimensional multifunctional approaches open new windows into intracellular architectures dynamics, well their underlying biophysical rules, far beyond diffraction limit.

Language: Английский

Citations

11
Lipid-Directed Covalent Labeling of Plasma Membranes for Long-Term Imaging, Barcoding and Manipulation of Cells DOI Creative Commons

Nathan Aknine,

Rémi Pelletier, Andrey S. Klymchenko

et al.

JACS Au, Journal Year: 2025, Volume and Issue: 5(2), P. 922 - 936

Published: Feb. 12, 2025

Fluorescent probes for cell plasma membranes (PM) generally exploit a noncovalent labeling mechanism, which constitutes fundamental limitation in multiple bioimaging applications. Here, we report concept of lipid-directed covalent PM, exploits transient binding to the lipid membrane surface generating high local dye concentration, thus favoring ligation random proximal proteins. This yielded fluorescent PM called MemGraft, are built (cyanine Cy3 or Cy5) bearing low-affinity anchor and reactive group: an activated ester maleimide. In contrast specially designed control dyes commercial Cy3-based labels amino thiol groups, MemGraft stain efficiently revealing crucial role combined with optimal reactivity overcome existing limitations probes, makes them compatible fixation, permeabilization, trypsinization, presence serum. The latter allows long-term tracking video imaging dynamics without signs phototoxicity. strategy also enables staining cocultured cells labeled different colors exchange probes. Moreover, combination MemGraft-Cy3 MemGraft-Cy5 at ratios enabled barcoding least 5 color codes, important visualizing populations cells. Ultimately, found that efficient biotinylation surface, opening path engineering manipulation.

Language: Английский

Citations

1
Functional Materials for Subcellular Targeting Strategies in Cancer Therapy: Progress and Prospects DOI

Yanxiang Cheng,

Zhen Qu, Jiang Qian

et al.

Advanced Materials, Journal Year: 2023, Volume and Issue: unknown

Published: Sept. 4, 2023

Neoadjuvant and adjuvant therapies have made significant progress in cancer treatment. However, tumor therapy still faces challenges due to the intrinsic heterogeneity of cancer, genomic instability, formation an immunosuppressive microenvironment. Functional materials possess unique biological properties such as long circulation times, tumor-specific targeting, immunomodulation. The combination functional with natural substances nanotechnology has led development smart biomaterials multiple functions, high biocompatibilities, negligible immunogenicities, which can be used for precise Recently, subcellular structure-targeting received particular attention various biomedical applications including diagnosis, sensing, imaging tumors drug delivery. Subcellular organelle-targeting precisely accumulate therapeutic agents organelles, considerably reduce threshold dosages agents, minimize drug-related side effects. This review provides a systematic comprehensive overview research organelle-targeted based on nanomaterials. Moreover, it explains prospects precision oncology. will serve excellent cutting-edge guide researchers field therapy.

Language: Английский

Citations

19
Fluorescent Probes Based on Charge and Proton Transfer for Probing Biomolecular Environment DOI Creative Commons
Vasyl G. Pivovarenko, Andrey S. Klymchenko

The Chemical Record, Journal Year: 2023, Volume and Issue: 24(2)

Published: Dec. 29, 2023

Abstract Fluorescent probes for sensing fundamental properties of biomolecular environment, such as polarity and hydration, help to study assembly lipids into biomembranes, interactions biomolecules imaging physiological state the cells. Here, we summarize major efforts in development based on two photophysical mechanisms: (i) an excited‐state intramolecular charge transfer (ICT), which is represented by fluorescent solvatochromic dyes that shift their emission band maximum a function environment hydration; (ii) proton (ESIPT), with particular focus 5‐membered cyclic systems, 3‐hydroxyflavones, because they exhibit dual sensitive environment. For both ICT ESIPT dyes, design biological applications are summarized. Thus, bearing amphiphilic anchors target lipid membranes report organization, while targeting ligands direct them specific organelles local The labels, amino acid nucleic analogues inserted enable monitoring membranes, proteins acids. While relatively simple robust environment‐sensitive probes, feature high information content due emission. They constitute powerful toolbox addressing multitude questions.

Language: Английский

Citations

13
PSL Chemical Biology Symposia: The Increasing Impact of Chemistry in Life Sciences DOI

Romain Sastourné‐Haletou,

Sacha Marynberg, Arthur Prudêncio de Araújo Pereira

et al.

ChemBioChem, Journal Year: 2025, Volume and Issue: unknown

Published: April 7, 2025

This symposium is the 6th Paris Sciences & Lettres (PSL) Chemical Biology meeting (2015, 2016, 2019, 2023, 2024, 2025) being held at Institut Curie. initiative originally started in 2013 de Chimie des Substances Naturelles (ICSN) Gif‐sur‐Yvette and was mostly focused on organic synthesis. It then exported Curie to cover a larger scope, before becoming official French meeting. year, around 200 participants had opportunity meet world leaders chemistry biology who described their latest innovations future trends covering topics as diverse prebiotic chemistry, activity‐based protein profiling, high‐resolution cell imaging, nanotechnologies, bio‐orthogonal metal ion signaling, ferroptosis, biocatalysis.

Language: Английский

Citations

0
Cyclic chalcone dye: solvatochromism, polarity sensitivity and theoretical calculations DOI

Yong-Cheng Feng,

Jun‐Yu Luan,

Nan Jiang

et al.

Journal of Molecular Structure, Journal Year: 2025, Volume and Issue: unknown, P. 141417 - 141417

Published: Jan. 1, 2025

Language: Английский

Citations

0
Research Progress in In Vitro Screening Techniques for Natural Antithrombotic Medicines DOI Creative Commons
Xinyang Liu, Lü Chen, Li Lin

et al.

Pharmaceuticals, Journal Year: 2025, Volume and Issue: 18(2), P. 137 - 137

Published: Jan. 21, 2025

Natural medicines play an indispensable role in treating thrombotic-related diseases and a thorough investigation of their material basis is crucial for medicine development. The rapid advancement medicine-active component screening technologies has paved new avenues studying natural medicines, holding significant theoretical practical value. This review focuses on the application progress multimodal technologies, including high-throughput screening, chip technology, molecular biology methods, fluorescence sensors, computational biology, anticoagulant medicines. aim to provide reference framework validating active components early these can swiftly assess safety efficacy accelerating development process reducing failure rate clinical trials. Nonetheless, overall mechanisms action correlation between chemical thrombotic remain challenging areas that require further in-depth exploration technological innovation.

Language: Английский

Citations

0
Look Beyond Plasma Membrane Biophysics: Revealing Considerable Variability of the Dipole Potential Between Plasma and Organelle Membranes of Living Cells DOI Open Access
Máté Szabó, Bence Cs. Szabo,

Kitti Kurtan

et al.

International Journal of Molecular Sciences, Journal Year: 2025, Volume and Issue: 26(3), P. 889 - 889

Published: Jan. 22, 2025

Due to the lack of measurement techniques suitable for examining compartments intact, living cells, membrane biophysics is almost exclusively investigated in plasma despite fact that its alterations intracellular organelles may also contribute disease pathogenesis. Here, we employ a novel, easy-to-use, confocal microscopy-based approach utilizing F66, an environment-sensitive fluorophore combination with fluorescent organelle markers and quantitative image analysis determine magnitude molecular order-related dipole potential various tumor neural cell lines. Our comparative demonstrates considerable variations be large enough modulate protein functions, inward decreasing gradient on route secretory/endocytic pathway (plasma >> lysosome > Golgi endoplasmic reticulum), whereas mitochondrial membranes are characterized by slightly larger than lysosomes. sensitive quantify biophysical properties selectively their and, therefore, identify affected therapeutic targets diseases associated lipid composition thus such as tumors, metabolic, neurodegenerative, or lysosomal storage disorders.

Language: Английский

Citations

0
Measuring plasma membrane fluidity using confocal microscopy DOI
Pablo Carravilla, Luca Andronico, Jan Schlegel

et al.

Nature Protocols, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 19, 2025

Language: Английский

Citations

0
Lipid-directed covalent fluorescent labeling of plasma membranes for long-term imaging, barcoding and manipulation of cells DOI Creative Commons

Nathan Aknine,

Rémi Pelletier, Andrey S. Klymchenko

et al.

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

Published: July 28, 2024

Abstract Fluorescent probes for cell plasma membrane (PM) are generally based on amphiphilic anchors that incorporate non-covalently into biomembranes. Therefore, they not compatible with fixation and permeabilization, presence of serum, or co-culture because their exchange the medium. Here, we report a concept lipid-directed covalent labeling PM, which exploits transient binding to lipid surface generating high local dye concentration, thus favoring ligation random proximal proteins. This yielded class fluorescent PM (MemGraft), where cyanine (Cy3 Cy5) bears at its two ends low-affinity anchor reactive group: an activated ester maleimide. We found MemGraft these groups provide efficient labelling, in contrast series control compounds, including commercial Cy3-based labels amino thiol groups, revealing crucial role combined reactivity maleimide groups. In conventional probes, non-covalent interactions, labelling approach is fixation, trypsinization serum. The latter allows long-term tracking video imaging dynamics without signs phototoxicity. strategy also enables staining co-cultured cells labelled different colors exchange. Moreover, combination ratios MemGraft-Cy3 MemGraft-Cy5 enabled barcoding least 5 color codes, important visualizing multiple populations. Ultimately, biotinylation surface, opening path engineering manipulation.

Language: Английский

Citations

2