Imaging of Extracellular Vesicles by Atomic Force Microscopy

Skliar, Mikhail; Chernyshev, Vasiliy S.

doi:10.3791/59254

Cited by 33 publications

(40 citation statements)

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“…Dehydrated samples were completely dried using a critical point dryer for at least 30 min. For the SPM analysis, the dehydrated surface analyzed using an AFM (Veeco Dimension 3100, VEECO) for an area of 5 × 5 μm with tapping mode in 512 lines at a scan rate of ∼0.7 Hz (Skliar & Chernyshev, 2019). An AppNano Si probe (ACT‐50, AppNano) was used for the analysis (rectangular‐shaped cantilever with nominal length = 125 μm and width = 30 μm; and a pyramidal‐shaped tip with height 14–16 μm, and tip radius < 10 nm, spring constant 13–77 N/m and f = 300 kHz).…”

Section: Methodsmentioning

confidence: 99%

Single‐vesicle imaging and co‐localization analysis for tetraspanin profiling of individual extracellular vesicles

Han

Kang

et al. 2021

J of Extracellular Vesicle

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Extracellular vesicles (EVs) are secreted nano-sized vesicles that contain cellular proteins, lipids, and nucleic acids. Although EVs are expected to be biologically diverse, current analyses cannot adequately characterize this diversity because most are ensemble methods that inevitably average out information from diverse EVs. Here we describe a single vesicle analysis, which directly visualizes marker expressions of individual EVs using a total internal-reflection microscopy and analyzes their co-localization to investigate EV subpopulations. The single-vesicle imaging and colocalization analysis successfully illustrated the diversity of EVs and revealed distinct patterns of tetraspanin expressions. Application of the analysis demonstrated similarities and dissimilarities between the EV fractions that had been acquired from different conventional EV isolation methods. The analysis method developed in this study will provide a new and reliable tool for investigating characteristics of single EVs, and the findings of the analysis might increase understanding of the characteristics of EVs.

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Section: Methodsmentioning

confidence: 99%

Single‐vesicle imaging and co‐localization analysis for tetraspanin profiling of individual extracellular vesicles

Han

Kang

et al. 2021

J of Extracellular Vesicle

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“…12 In contrast, AFM offers the ability to measure label-free samples in their native conditions with minimal sample preparation whilst enabling the high-resolution analysis necessary to investigate EV structure. 13 AFM measures the interaction between a probing tip and the sample surface and is able to determine EV size distribution, concentration and morphology. AFM can also map biomechanical properties including surface roughness, membrane elasticity and both non-specific, and ligand-specific surface adhesion via antibody/ligand coated tips.…”

Section: Introductionmentioning

confidence: 99%

High content, quantitative AFM analysis of the scalable biomechanical properties of extracellular vesicles

et al. 2021

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“…These results suggest a stable suspension. Figure 2 B,D shows representative AFM topography images displaying particles with a pancake-like morphology, which is characteristic of soft particles adsorbed on a surface [ 25 ]. A low amount of organic material was covering the substrate and surrounding the particles, as can be seen in the phase image from Figure 2 C. Therefore, line profiles of areas where the particles were adsorbed, as indicated in Figure 2 D, indicated particles that were between 3 nm and 40 nm in height, according to Figure 2 E.…”

Section: Resultsmentioning

confidence: 99%

Increased Fibroblast Metabolic Activity of Collagen Scaffolds via the Addition of Propolis Nanoparticles

et al. 2020

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Propolis natural extracts have been used since ancient times due to their antioxidant, anti-inflammatory, antiviral, and antimicrobial activities. In this study, we produced scaffolds of type I collagen, extracted from Wistar Hanover rat tail tendons, and impregnated them with propolis nanoparticles (NPs) for applications in regenerative medicine. Our results show that the impregnation of propolis NPs to collagen scaffolds affected the collagen denaturation temperature and tensile strength. The changes in structural collagen self-assembly due to contact with organic nanoparticles were shown for the first time. The fibril collagen secondary structure was preserved, and the D-pattern gap increased to 135 ± 28 nm, without losing the microfiber structure. We also show that the properties of the collagen scaffolds depended on the concentration of propolis NPs. A concentration of 100 μg/mL of propolis NPs with 1 mg of collagen, with a hydrodynamic diameter of 173 nm, was found to be an optimal concentration to enhance 3T3 fibroblast cell metabolic activity and cell proliferation. The expected outcome from this research is both scientifically and socially relevant since the home scaffold using natural nanoparticles can be produced using a simple method and could be widely used for local medical care in developing communities.

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Imaging of Extracellular Vesicles by Atomic Force Microscopy

Cited by 33 publications

References 0 publications

Single‐vesicle imaging and co‐localization analysis for tetraspanin profiling of individual extracellular vesicles

Single‐vesicle imaging and co‐localization analysis for tetraspanin profiling of individual extracellular vesicles

High content, quantitative AFM analysis of the scalable biomechanical properties of extracellular vesicles

Increased Fibroblast Metabolic Activity of Collagen Scaffolds via the Addition of Propolis Nanoparticles

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