Resolving the nanoscale structure of β-sheet assemblies using single-molecule orientation-localization microscopy DOI Creative Commons
Weiyan Zhou, Conor L. O’Neill, Tianben Ding

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Sept. 14, 2023

Abstract Synthetic peptides that self-assemble into cross-β fibrils have remarkable utility as engineered biomaterials due to their modularity and biocompatibility, but structural morphological similarity amyloid species has been a long-standing concern for translation. Further, polymorphs are difficult characterize using spectroscopic imaging techniques rely on ensemble averaging achieve high resolution. Here, we utilize single-molecule orientation-localization microscopy (SMOLM) formed by the designed amphipathic enantiomers, KFE8 L D , pathological amyloid-beta peptide Aβ42. SMOLM reveals orientations of Nile red, it transiently binds both Aβ42, consistent with helical (bilayer) ribbon structure convey precise tilt fibrils’ inner outer backbones. also finds polymorphic branched curved morphologies whose backbones exhibit much more heterogeneity than those typical straight fibrils. Thus, is powerful tool interrogate differences polymorphism between cross β-rich

Language: Английский

The Advances and Applications of Characterization Technique for Exosomes: From Dynamic Light Scattering to Super-Resolution Imaging Technology DOI Creative Commons
Shijia Wu,

Yalan Zhao,

Zitong Zhang

et al.

Photonics, Journal Year: 2024, Volume and Issue: 11(2), P. 101 - 101

Published: Jan. 23, 2024

Exosomes distributed by extracellular vesicles carry various information highly consistent with cells, becoming a new type of biomarker for tumor screening. However, although conventional characterization technologies can quantify size and morphology exosomes, they are limited in related fields such as function tracing, protein quantification at unit point, microstructural information. In this paper, firstly, different exosome methods systematically reviewed, dynamic light scattering, nanoparticle tracking analysis, flow cytometry, electron microscope, emerging super-resolution imaging technologies. Then, advances applications described one one. Last but not least, we compare the features exosomes propose that technology only take into account advantages techniques also provide accurate, real-time, quantitative analysis exosomes. It provides fine guide exosome-related biomedical research, well application liquid biopsy techniques.

Language: Английский

Citations

15
Continuous Monitoring Biosensing Mediated by Single-Molecule Plasmon-Enhanced Fluorescence in Complex Matrices DOI Creative Commons

Vincenzo Lamberti,

Mathias Dolci, Peter Zijlstra

et al.

ACS Nano, Journal Year: 2024, Volume and Issue: unknown

Published: Feb. 9, 2024

Continuous detection of critical markers directly at the point interest and in undiluted biological fluids represents next fundamental step biosensing. The goal realizing such a platform is utterly challenging because it requires reversible biosensor that enables tracking pico- to nanomolar molecular concentrations over long time spans compact device. Here we describe sensing method based on plasmon-enhanced fluorescence capable single-molecule unlabeled analyte by employing biofunctionalized gold nanoparticles. very strong signals allow for unaltered media, while use low-affinity interactions ensures continuous increasing decreasing with picomolar sensitivity. We demonstrate sandwich assay DNA cancer marker limit response 10 min. enhanced will miniaturization into small cheap multiplexing capability application point-of-care diagnostics, monitoring industrial processes, safe keeping environment.

Language: Английский

Citations

9
Resolving the Nanoscale Structure of β-Sheet Peptide Self-Assemblies Using Single-Molecule Orientation–Localization Microscopy DOI
Weiyan Zhou, Conor L. O’Neill, Tianben Ding

et al.

ACS Nano, Journal Year: 2024, Volume and Issue: 18(12), P. 8798 - 8810

Published: March 13, 2024

Synthetic peptides that self-assemble into cross-β fibrils are versatile building blocks for engineered biomaterials due to their modularity and biocompatibility, but structural morphological similarities amyloid species have been a long-standing concern translation. Further, polymorphs difficult characterize by using spectroscopic imaging techniques rely on ensemble averaging achieve high resolution. Here, we utilize Nile red (NR), an amyloidophilic fluorogenic probe, single-molecule orientation–localization microscopy (SMOLM) formed the designed amphipathic enantiomers KFE8L KFE8D pathological amyloid-beta peptide Aβ42. Importantly, NR SMOLM reveals helical (bilayer) ribbon structure of both KFE8 Aβ42 quantifies precise tilt fibrils' inner outer backbones in relevant buffer conditions without need covalent labeling or sequence mutations. also distinguishes polymorphic branched curved morphologies KFE8, whose exhibit much more heterogeneity than those typical straight fibrils. Thus, is powerful tool interrogate differences polymorphism between cross-β-rich

Language: Английский

Citations

5
Single-molecule orientation-localization microscopy: Applications and approaches DOI
Oumeng Zhang, Matthew D. Lew

Quarterly Reviews of Biophysics, Journal Year: 2024, Volume and Issue: 57

Published: Jan. 1, 2024

Abstract Single-molecule orientation-localization microscopy (SMOLM) builds upon super-resolved localization by imaging orientations and rotational dynamics of individual molecules in addition to their positions. This added dimensionality provides unparalleled insights into nanoscale biophysical biochemical processes, including the organization actin networks, movement molecular motors, conformations DNA strands, growth remodeling amyloid aggregates, composition changes within lipid membranes. In this review, we discuss recent innovations SMOLM cover three key aspects: (1) enabled labeling strategies that endow fluorescent probes bind targets with orientation specificity; (2) advanced techniques leverage physics light-matter interactions estimation theory encode information high fidelity microscope images; (3) computational methods ensure accurate precise data analysis interpretation, even presence severe shot noise. Additionally, compare approaches, hardware, publicly available software aid community choosing best implementation for specific application. Finally, highlight future directions SMOLM, such as development improved photostability specificity, design “smart” adaptive use approaches handle large, complex datasets. review underscores significant current potential impact deepening our understanding dynamics, paving way breakthroughs fields biophysics, biochemistry, materials science.

Language: Английский

Citations

5
Advanced optical imaging for the rational design of nanomedicines DOI Creative Commons
Ana Ortiz‐Perez, Miao Zhang, Laurence W. Fitzpatrick

et al.

Advanced Drug Delivery Reviews, Journal Year: 2023, Volume and Issue: 204, P. 115138 - 115138

Published: Nov. 18, 2023

Despite the enormous potential of nanomedicines to shape future medicine, their clinical translation remains suboptimal. Translational challenges are present in every step development pipeline, from a lack understanding patient heterogeneity insufficient insights on nanoparticle properties and impact material-cell interactions. Here, we discuss how adoption advanced optical microscopy techniques, such as super-resolution microscopies, correlative high-content modalities, could aid rational design nanocarriers, by characterizing cell, nanomaterial, interaction with unprecedented spatial and/or temporal detail. In this nanomedicine arena, will implementation these versatility specificity, can yield high volumes multi-parametric data; machine learning rapid advances microscopy: image acquisition data interpretation.

Language: Английский

Citations

11
Glyco-PAINT-APP: Subcellular analysis of immune cell-lectin binding enables correlation of glycan binding parameters to receptor biology DOI Creative Commons

Kas Steuten,

Jaap Bakker, Ward Doelman

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 24, 2025

Abstract Extracting single-molecule glycan binding data from primary cells has not been possible to date. Here, we present Glyco-PAINT-APP (Automated Processing Pipeline), an automated method that enables the extraction of subcellular interaction kinetics using a Points Accumulation for Imaging in Nano-Topography (PAINT)-based approach. This approach leverages algorithm precise, high-throughput analysis dynamics, facilitating exploration functional correlations between glycoform patterns and immune cell polarization. Using synthetic glycans glycosylated antigens, demonstrate ability technique automatically correlate high regions dendritic with increased uptake cross-presentation efficiency these antigens. Additionally, show how can uncover subtle differences preferences resting polarized macrophages. Taken together, potential enable new insights into cell-intrinsic heterogeneity glycan-structure-activity relationships cells.

Language: Английский

Citations

0
Characterisation of polymeric nanoparticles for drug delivery DOI Creative Commons

Thomas G. Floyd,

Pratik Gurnani, Julia Y. Rho

et al.

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

Published: Jan. 1, 2025

Polymeric nanoparticles represent an innovative approach to drug delivery, particularly for addressing complex diseases like cancer.

Language: Английский

Citations

0
The integration of nanotechnology, nanomedicine, and artificial intelligence for advancements in healthcare: a Conceptual Review Based on PRISMA Method and Future Research Directions DOI
Piumika Yapa, Sisitha Rajapaksha, Imalka Munaweera

et al.

Next research., Journal Year: 2025, Volume and Issue: unknown, P. 100330 - 100330

Published: April 1, 2025

Language: Английский

Citations

0
Advancements in Single-Molecule Fluorescence Detection Techniques and Their Expansive Applications in Drug Discovery and Neuroscience DOI Creative Commons

Jing Yan,

Lin Cheng, Yitong Li

et al.

Biosensors, Journal Year: 2025, Volume and Issue: 15(5), P. 283 - 283

Published: April 30, 2025

Single-molecule fluorescence technology stands at the forefront of scientific research as a sophisticated tool, pushing boundaries our understanding. This review comprehensively summarizes technological advancements in single-molecule detection, highlighting latest achievements development fluorescent probes, imaging systems, and biosensors. It delves into applications these cutting-edge tools drug discovery neuroscience research, encompassing design monitoring complex delivery elucidation pharmacological mechanisms pharmacokinetics, intricacies neuronal signaling synaptic function, molecular underpinnings neurodegenerative diseases. The exceptional sensitivity demonstrated underscores vast potential modern biomedical heralding its expansion other domains.

Language: Английский

Citations

0
Super-resolution microscopy to study membrane nanodomains and transport mechanisms in the plasma membrane DOI Creative Commons
Yenisleidy de las Mercedes Zulueta Díaz, Eva C. Arnspang

Frontiers in Molecular Biosciences, Journal Year: 2024, Volume and Issue: 11

Published: Sept. 3, 2024

Biological membranes are complex, heterogeneous, and dynamic systems that play roles in the compartmentalization protection of cells from environment. It is still a challenge to elucidate kinetics real-time transport routes for molecules through biological live cells. Currently, by developing employing super-resolution microscopy; increasing evidence indicates channels transporter nano-organization dynamics within an important role these regulatory mechanisms. Here we review recent advances discuss major advantages disadvantages using microscopy investigate protein organization plasma membranes.

Language: Английский

Citations

3