scholarly journals AFM-based High-Throughput Nanomechanical Screening of Single Extracellular Vesicles

2019 ◽  
Author(s):  
Andrea Ridolfi ◽  
Marco Brucale ◽  
Costanza Montis ◽  
Lucrezia Caselli ◽  
Lucia Paolini ◽  
...  

AbstractWe herein describe an Atomic Force Microscopy (AFM)-based experimental procedure which allows the simultaneous mechanical and morphological characterization of several hundred individual nanosized vesicles within the hour timescale.When deposited on a flat rigid surface from aqueous solution, vesicles are deformed by adhesion forces into oblate spheroids whose geometry is a direct consequence of their mechanical stiffness. AFM image analysis can be used to quantitatively measure the contact angle of individual vesicles, which is a size-independent descriptor of their deformation and, consequently, of their stiffness. The same geometrical measurements can be used to infer vesicle diameter in its original, spherical shape.We demonstrate the applicability of the proposed approach to natural vesicles obtained from different sources, recovering their size and stiffness distributions by simple AFM imaging in liquid. We show how the combined EV stiffness/size readout is able to discriminate between subpopulations of vesicular and non-vesicular objects in the same sample, and between populations of vesicles with similar sizes but different mechanical characteristics. We also discuss a force spectroscopy calibration procedure to quantitatively link the stiffness of EVs to their average contact angle.Finally, we discuss expected extensions and applications of the methodology.

2017 ◽  
Vol 32 (6) ◽  
pp. 813-825 ◽  
Author(s):  
Bipin Gaihre ◽  
Beata Lecka-Czernik ◽  
Ambalangodage C Jayasuriya

This study was aimed at assessing the effects of silica nanopowder incorporation into chitosan-tripolyphosphate microparticles with the ultimate goal of improving their osteogenic properties. The microparticles were prepared by simple coacervation technique and silica nanopowder was added at 0% (C), 2.5% (S1), 5% (S2) and 10% (S3) (w/w) to chitosan. We observed that this simple incorporation of silica nanopowder improved the growth and proliferation of osteoblasts along the surface of the microparticles. In addition, the composite microparticles also showed the increased expression of alkaline phosphatase and osteoblast specific genes. We observed a significant increase ( p < 0.05) in the expression of alkaline phosphatase by the cells growing on all sample groups compared to the control (C) groups at day 14. The morphological characterization of these microparticles through scanning electron microscopy showed that these microparticles were well suited to be used as the injectable scaffolds with perfectly spherical shape and size. The incorporation of silica nanopowder altered the nano-roughness of the microparticles as observed through atomic force microscopy scans with roughness values going down from C to S3. The results in this study, taken together, show the potential of chitosan-tripolyphosphate-silica nanopowder microparticles for improved bone regeneration applications.


2019 ◽  
Vol 60 ◽  
pp. 124-141 ◽  
Author(s):  
Naser Ali ◽  
Joao Amaral Teixeira ◽  
Abdulmajid Addali

This research investigates the effect of surface roughness, water temperature, and pH value on the wettability behaviour of copper surfaces. An electron beam physical vapour deposition technique was used to fabricate 25, 50, and 75 nm thin films of copper on the surface of copper substrates. Surface topographical analysis, of the uncoated and coated samples, was performed using an atomic force microscopy device to observe the changes in surface microstructure. A goniometer device was then employed to examine the surface wettability of the samples by obtaining the static contact angle between the liquid and the attached surface using the sessile drops technique. Waters of pH 4, 7, and 9 were employed as the contact angle testing fluids at a set of fixed temperatures that ranged from 20°C to 60°C. It was found that increasing the deposited film thickness reduces the surface roughness of the as-prepared copper surfaces and thus causing the surface wettability to diverge from its initial hydrophobic nature towards the hydrophilic behaviour region. A similar divergence behaviour was seen with the rise in temperature of water of pH 4, and 9. In contrast, the water of pH 7, when tested on the uncoated surface, ceased to reach a contact angle below 90o. It is believed that the observed changes in surface wettability behaviour is directly linked to the liquid temperature, pH value, surface roughness, along with the Hofmeister effect between the water and the surface in contact.


2007 ◽  
Vol 342-343 ◽  
pp. 221-224
Author(s):  
Jin Suk Bae ◽  
Ga Young Jun ◽  
Akihiko Kikuchi ◽  
Teruo Okano ◽  
Chang Hyun Ahn ◽  
...  

In this work, we developed a novel patterned co-culture method with thermo-responsive poly(N-isopropylacrylamide) (PIPAAm) and poly(N-ρ-vinylbenzyl-Ο-β-D-galactopyranosyl-(1→ 4)-D-gluconamide) (PVLA) inducing active hepatocyte attachment. Patterned graft of PIPAAm onto PS dishes was carried out by electron beam irradiation using cover-glass as a photomask. PVLA was only coated onto PIPAAm-ungrafted domain because of hydrated hydrophilic property of PIPAAm at below the LCST. Analysis by attenuated total reflection-Fourier transform infrared and electron spectroscopy for chemical analysis revealed that PIPAAm and PVLA were successfully grafted and coated on surfaces of PS dishes. PIPAAm-grafted surface exhibited decreasing contact angle by changing temperature from 37 to 20°C, while PVLA-coated PS and non-treated PS had negligible contact angle changes with temperature alternation. Atomic force microscopy (AFM) results showed that PIPAAm-grafted and PVLA-coated PS had smoother surfaces than that of ungrafted PS dishes. After culture for 12 hours, hepatocytes were well attached on PVLA-coated domain. Hepatocytes adherent on PIPAAm-grafted domain were detached by decreasing temperature. And then, fibroblasts were seeded onto PIPAAm pattern-grafted domain. Fibroblasts were only attached and spread onto PIPAAm-grafted domain. Co-cultured hepatocytes showed better differentiated function of albumin expression compared to homotypic hepatocyte culture


2016 ◽  
Vol 1 (1) ◽  
pp. 69-74 ◽  
Author(s):  
Sofiane El-Kirat-Chatel ◽  
Yves F. Dufrêne

We establish atomic force microscopy as a new nanoscopy platform for quantifying the forces between fungal pathogens and immune cells.


1996 ◽  
Vol 440 ◽  
Author(s):  
H. C. Wang ◽  
D. W. Cheong ◽  
J. Kumar ◽  
C. Sung ◽  
S. K. Tripathy

AbstractA soluble, asymmetrically substituted polydiacetylene, poly(BPOD), has been reported to form stable monolayers at the air-water interface by the Langmuir-Blodgett (LB) technique [2]. Preformed polydiacetylene has been deposited onto hydrophobic substrates as multilayers to form second order nonlinear optical thin films. Second harmonic generation was found to increase with the number of layers. From previous atomic force microscopy (AFM) studies backbone orientation along the dipping direction with an interchain spacing of about 5 A° was indicated [2].The film morphology and preferential molecular orientation of these LB films are further investigated by transmission electron microscopy (TEM). A specifically tailored sample preparation method for the ultrathin LB films was used. Multilayer films were deposited on hydrophobic collodion covered glass substrates for this purpose. Electron diffraction was employed to study the crystalline organization of mono and multilayers of LB films as well as cast films.


Materials ◽  
2020 ◽  
Vol 13 (14) ◽  
pp. 3171
Author(s):  
AbdolAli Moghaddasi ◽  
Patrik Sobolčiak ◽  
Anton Popelka ◽  
Igor Krupa

Purpose: Copolyamide 6,10 (coPA) electrospun mats were covered with multilayered (ML) and single-layered (SL) MXene (Ti3C2Tx) as a membrane for the separation of water/vegetable oil emulsions. Methods: Prepared membranes were characterized by atomic force microscopy (AFM), profilometry, the contact angle measurements of various liquids in air, and the underwater contact angle of vegetable oil. The separation efficiency was evaluated by measuring the UV transmittance of stock solutions compared to the UV transmittance of the filtrate. Results: The MXene coating onto coPA mats led to changes in the permeability, hydrophilicity, and roughness of the membranes and enhanced the separation efficiency of the water/vegetable oil emulsions containing 10, 100, and 1000 ppm of sunflower vegetable oil. It was found that membranes were highly oleophobic (>124°) under water, unlike in air, where the membranes showed high oleophobicity (<5°). The separation efficiency of water/oil emulsions for both types of covered membranes reached over 99%, with a surface coverage of 3.2 mg/cm2 Ti3C2Tx (for ML-Ti3C2Tx) and 2.9 mg/cm2 (for SL-Ti3C2Tx). Conclusions: The separation efficiency was greater than 98% for membranes covered with 2.65 mg/cm2 of ML-Ti3C2Tx, whereas the separation efficiency for membranes containing 1.89 and 0.77 mg/cm2 was less than 90% for all studied emulsion concentrations.


Coatings ◽  
2020 ◽  
Vol 10 (7) ◽  
pp. 659 ◽  
Author(s):  
Marcela Elisabeta Barbinta-Patrascu ◽  
Camelia Ungureanu ◽  
Nicoleta Badea ◽  
Mihaela Bacalum ◽  
Andrada Lazea-Stoyanova ◽  
...  

The objective of the present study is the valorization of natural resources and the recycling of vegetal wastes by converting them into novel plasmonic bio-active hybrids. Thus, a “green” approach was used to design pectin-coated bio-nanosilver. Silver nanoparticles were generated from two common garden herbs (Mentha piperita and Amaranthus retroflexus), and pectin was extracted from lemon peels. The samples were characterized by the following methods: Ultraviolet–visible (UV-Vis) absorption spectroscopy, Fourier Transform Infrared (FT-IR), X-ray Diffraction (XRD), X-ray Photoelectron Spectroscopy (XPS), dynamic light scattering (DLS), Atomic Force Microscopy (AFM), Scanning Electron Microscopy (SEM)–Energy-dispersive X-ray Spectroscopy (EDX), and zeta potential measurements. Microscopic investigations revealed the spherical shape and the nano-scale size of the prepared biohybrids. Their bioperformances were checked in terms of antioxidant and antibacterial activity. The developed plasmonic materials exhibited a strong ability to scavenge short-life (96.1% ÷ 98.7%) and long-life (39.1% ÷ 91%) free radicals. Microbiological analyses demonstrated an impressive antibacterial effectiveness of pectin-based hybrids against Escherichia coli. The results are promising, and the obtained biomaterials could be used in many bio-applications, especially as antioxidant and antimicrobial biocoatings.


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