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1.
Nature ; 2024 Jun 05.
Article in English | MEDLINE | ID: mdl-38839964

ABSTRACT

Membranes are widely used for separation processes in applications such as water desalination, batteries and dialysis, and are crucial in key sectors of our economy and society1. The majority of technologically exploited membranes are based on solid polymers and function as passive barriers, whose transport characteristics are governed by their chemical composition and nanostructure. Although such membranes are ubiquitous, it has proved challenging to maximize selectivity and permeability independently, leading to trade-offs between these pertinent characteristics2. Self-assembled biological membranes, in which barrier and transport functions are decoupled3,4, provide the inspiration to address this problem5,6. Here we introduce a self-assembly strategy that uses the interface of an aqueous two-phase system to template and stabilize molecularly thin (approximately 35 nm) biomimetic block copolymer bilayers of scalable area that can exceed 10 cm2 without defects. These membranes are self-healing, and their barrier function against the passage of ions (specific resistance of approximately 1 MΩ cm2) approaches that of phospholipid membranes. The fluidity of these membranes enables straightforward functionalization with molecular carriers that shuttle potassium ions down a concentration gradient with exquisite selectivity over sodium ions. This ion selectivity enables the generation of electric power from equimolar solutions of NaCl and KCl in devices that mimic the electric organ of electric rays.

2.
J Vis Exp ; (205)2024 Mar 22.
Article in English | MEDLINE | ID: mdl-38587397

ABSTRACT

High-speed atomic force microscopy (HS-AFM) is a popular molecular imaging technique for visualizing single-molecule biological processes in real-time due to its ability to image under physiological conditions in liquid environments. The photothermal off-resonance tapping (PORT) mode uses a drive laser to oscillate the cantilever in a controlled manner. This direct cantilever actuation is effective in the MHz range. Combined with operating the feedback loop on the time domain force curve rather than the resonant amplitude, PORT enables high-speed imaging at up to ten frames per second with direct control over tip-sample forces. PORT has been shown to enable imaging of delicate assembly dynamics and precise monitoring of patterns formed by biomolecules. Thus far, the technique has been used for a variety of dynamic in vitro studies, including the DNA 3-point-star motif assembly patterns shown in this work. Through a series of experiments, this protocol systematically identifies the optimal imaging parameter settings and ultimate limits of the HS-PORT AFM imaging system and how they affect biomolecular assembly processes. Additionally, it investigates potential undesired thermal effects induced by the drive laser on the sample and surrounding liquid, particularly when the scanning is limited to small areas. These findings provide valuable insights that will drive the advancement of PORT mode's application in studying complex biological systems.


Subject(s)
Mechanical Phenomena , Nanotechnology , Microscopy, Atomic Force/methods , Molecular Imaging , DNA
3.
Beilstein J Nanotechnol ; 15: 134-143, 2024.
Article in English | MEDLINE | ID: mdl-38317825

ABSTRACT

Dynamic atomic force microscopy (AFM) modes that operate at frequencies far away from the resonance frequency of the cantilever (off-resonance tapping (ORT) modes) can provide high-resolution imaging of a wide range of sample types, including biological samples, soft polymers, and hard materials. These modes offer precise and stable control of vertical force, as well as reduced lateral force. Simultaneously, they enable mechanical property mapping of the sample. However, ORT modes have an intrinsic drawback: a low scan speed due to the limited ORT rate, generally in the low-kilohertz range. Here, we analyze how the conventional ORT control method limits the topography tracking quality and hence the imaging speed. The closed-loop controller in conventional ORT restricts the sampling rate to the ORT rate and introduces a large closed-loop delay. We present an alternative ORT control method in which the closed-loop controller samples and tracks the vertical force changes during a defined time window of the tip-sample interaction. Through this, we use multiple samples in the proximity of the maximum force for the feedback loop, rather than only one sample at the maximum force instant. This method leads to improved topography tracking at a given ORT rate and therefore enables higher scan rates while refining the mechanical property mapping.

4.
Microsyst Nanoeng ; 10: 28, 2024.
Article in English | MEDLINE | ID: mdl-38405129

ABSTRACT

Grayscale structured surfaces with nanometer-scale features are used in a growing number of applications in optics and fluidics. Thermal scanning probe lithography achieves a lateral resolution below 10 nm and a vertical resolution below 1 nm, but its maximum depth in polymers is limited. Here, we present an innovative combination of nanowriting in thermal resist and plasma dry etching with substrate cooling, which achieves up to 10-fold amplification of polymer nanopatterns into SiO2 without proportionally increasing surface roughness. Sinusoidal nanopatterns in SiO2 with 400 nm pitch and 150 nm depth are fabricated free of shape distortion after dry etching. To exemplify the possible applications of the proposed method, grayscale dielectric nanostructures are used for scalable manufacturing through nanoimprint lithography and for strain nanoengineering of 2D materials. Such a method for aspect ratio amplification and smooth grayscale nanopatterning has the potential to find application in the fabrication of photonic and nanoelectronic devices.

6.
STAR Protoc ; 4(3): 102468, 2023 Sep 15.
Article in English | MEDLINE | ID: mdl-37481726

ABSTRACT

Atomic force microscopy (AFM) is capable of nanoscale imaging but has so far only been used on cell surfaces when applied to a living cell. Here, we describe a step-by-step protocol for nanoendoscopy-AFM, which enables the imaging of nanoscale structures inside living cells. The protocol consists of cell staining, fabrication of the nanoneedle probes, observation inside living cells using 2D and 3D nanoendoscopy-AFM, and visualization of the 3D data. For complete details on the use and execution of this protocol, please refer to Penedo et al. (2021)1 and Penedo et al. (2021).2.


Subject(s)
Nanotechnology , Microscopy, Atomic Force/methods , Nanotechnology/methods , Cell Membrane/chemistry
7.
J Phys Chem Lett ; : 5365-5371, 2022 Jun 09.
Article in English | MEDLINE | ID: mdl-35678499

ABSTRACT

Three-dimensional atomic force microscopy (3D-AFM) has resolved three-dimensional distributions of solvent molecules at solid-liquid interfaces at the subnanometer scale. This method is now being extended to the imaging of biopolymer assemblies such as chromosomes or proteins in cells, with the expectation of being able to resolve their three-dimensional structures. Here, we have developed a computational method to simulate 3D-AFM images of biopolymers by using the Jarzynski equality. It is found that some parts of the fiber structure of biopolymers are indeed resolved in the 3D-AFM image. The dependency of 3D-AFM images on the vertical scanning velocity is investigated, and optimum scanning velocities are found. It is also clarified that forces in nonequilibrium processes are measured in 3D-AFM measurements when the dynamics of polymers are slower than the scanning of the probe.

8.
Sci Adv ; 7(52): eabj4990, 2021 Dec 24.
Article in English | MEDLINE | ID: mdl-34936434

ABSTRACT

Atomic force microscopy (AFM) is the only technique that allows label-free imaging of nanoscale biomolecular dynamics, playing a crucial role in solving biological questions that cannot be addressed by other major bioimaging tools (fluorescence or electron microscopy). However, such imaging is possible only for systems either extracted from cells or reconstructed on solid substrates. Thus, nanodynamics inside living cells largely remain inaccessible with the current nanoimaging techniques. Here, we overcome this limitation by nanoendoscopy-AFM, where a needle-like nanoprobe is inserted into a living cell, presenting actin fiber three-dimensional (3D) maps, and 2D nanodynamics of the membrane inner scaffold, resulting in undetectable changes in cell viability. Unlike previous AFM methods, the nanoprobe directly accesses the target intracellular components, exploiting all the AFM capabilities, such as high-resolution imaging, nanomechanical mapping, and molecular recognition. These features should greatly expand the range of intracellular structures observable in living cells.

9.
Sci Rep ; 11(1): 7756, 2021 04 08.
Article in English | MEDLINE | ID: mdl-33833307

ABSTRACT

Over the last decade, nanoneedle-based systems have demonstrated to be extremely useful in cell biology. They can be used as nanotools for drug delivery, biosensing or biomolecular recognition inside cells; or they can be employed to select and sort in parallel a large number of living cells. When using these nanoprobes, the most important requirement is to minimize the cell damage, reducing the forces and indentation lengths needed to penetrate the cell membrane. This is normally achieved by reducing the diameter of the nanoneedles. However, several studies have shown that nanoneedles with a flat tip display lower penetration forces and indentation lengths. In this work, we have tested different nanoneedle shapes and diameters to reduce the force and the indentation length needed to penetrate the cell membrane, demonstrating that ultra-thin and sharp nanoprobes can further reduce them, consequently minimizing the cell damage.


Subject(s)
Microscopy, Atomic Force/methods , Nanotechnology , Needles , Cell Separation
10.
Nat Commun ; 11(1): 6365, 2020 Dec 11.
Article in English | MEDLINE | ID: mdl-33311480

ABSTRACT

Materials hosting magnetic skyrmions at room temperature could enable compact and energetically-efficient storage such as racetrack memories, where information is coded by the presence/absence of skyrmions forming a moving chain through the device. The skyrmion Hall effect leading to their annihilation at the racetrack edges can be suppressed, for example, by antiferromagnetically-coupled skyrmions. However, avoiding modifications of the inter-skyrmion distances remains challenging. As a solution, a chain of bits could also be encoded by two different solitons, such as a skyrmion and a chiral bobber, with the limitation that it has solely been realized in B20-type materials at low temperatures. Here, we demonstrate that a hybrid ferro/ferri/ferromagnetic multilayer system can host two distinct skyrmion phases at room temperature, namely tubular and partial skyrmions. Furthermore, the tubular skyrmion can be converted into a partial skyrmion. Such systems may serve as a platform for designing memory applications using distinct skyrmion types.

11.
Sci Rep ; 10(1): 17436, 2020 10 15.
Article in English | MEDLINE | ID: mdl-33060692

ABSTRACT

In recent years, the atomic force microscope has proven to be a powerful tool for studying biological systems, mainly for its capability to measure in liquids with nanoscale resolution. Measuring tissues, cells or proteins in their physiological conditions gives us access to valuable information about their real 'in vivo' structure, dynamics and functionality which could then fuel disruptive medical and biological applications. The main problem faced by the atomic force microscope when working in liquid environments is the difficulty to generate clear cantilever resonance spectra, essential for stable operation and for high resolution imaging. Photothermal actuation overcomes this problem, as it generates clear resonance spectra free from spurious peaks. However, relatively high laser powers are required to achieve the desired cantilever oscillation amplitude, which could potentially damage biological samples. In this study, we demonstrate that the photothermal excitation efficiency can be enhanced by coating the cantilever with a thin amorphous carbon layer to increase the heat absorption from the laser, reducing the required excitation laser power and minimizing the damage to biological samples.

12.
Nano Lett ; 18(4): 2263-2267, 2018 04 11.
Article in English | MEDLINE | ID: mdl-29543463

ABSTRACT

Mallinson's idea that some spin textures in planar magnetic structures could produce an enhancement of the magnetic flux on one side of the plane at the expense of the other gave rise to permanent magnet configurations known as Halbach magnet arrays. Applications range from wiggler magnets in particle accelerators and free electron lasers to motors and magnetic levitation trains, but exploiting Halbach arrays in micro- or nanoscale spintronics devices requires solving the problem of fabrication and field metrology below a 100 µm size. In this work, we show that a Halbach configuration of moments can be obtained over areas as small as 1 µm × 1 µm in sputtered thin films with Néel-type domain walls of unique domain wall chirality, and we measure their stray field at a controlled probe-sample distance of 12.0 ± 0.5 nm. Because here chirality is determined by the interfacial Dyzaloshinkii-Moriya interaction, the field attenuation and amplification is an intrinsic property of this film, allowing for flexibility of design based on an appropriate definition of magnetic domains. Skyrmions (<100 nm wide) illustrate the smallest kind of such structures, for which our measurement of stray magnetic fields and mapping of the spin structure shows they funnel the field toward one specific side of the film given by the sign of the Dyzaloshinkii-Moriya interaction parameter D.

13.
Nanotechnology ; 26(48): 485706, 2015 Dec 04.
Article in English | MEDLINE | ID: mdl-26559931

ABSTRACT

Multifrequency atomic force microscopy (AFM) in liquid media where several eigenmodes or harmonics are simultaneously excited is improving the performance of the scanning probe techniques in biological studies. As a consequence, an important effort is being made to search for a reliable, efficient and strong cantilever high mode excitation method that operates in liquids. In this work we present (theoretical and experimentally) a technique for improving the efficiency of the most common excitation methods currently used in AFM operated in liquids: photothermal, torque (MAC Mode™) and magnetostriction. By etching specific areas of the cantilever coating, the oscillation amplitude (both flexural and torsional) of each specific eigenmode increases, leading to an improvement in signal to noise ratio of the multifrequency techniques. As an alternative, increment in high mode oscillation amplitude is also obtained by Ga(+) ion implantation in the specific areas of the magnetic material.

14.
Nanotechnology ; 24(39): 395701, 2013 Oct 04.
Article in English | MEDLINE | ID: mdl-24008394

ABSTRACT

The tip apex dimensions and geometry of the conductive probe remain the major limitation to the resolution of Kelvin probe force microscopy (KPFM). One of the possible strategies to improve the spatial resolution of surface potential images consists in the development of thinner and more durable conductive tips. In an effort to improve the lateral resolution of topography and surface potential maps, we have evaluated high aspect ratio conductive tips created by depositing gold nanoparticles on standard silicon tips. Besides the already known general topographic resolution enhancement offered by these modified tips, an improvement of surface potential lateral resolution and signal-to-noise ratio is reported here for a variety of samples as compared to other regular conductive probes. We have also observed that the modified conductive tips have a significant auto-regeneration capability, which stems from a certain level of mobility of the nanoparticle coating. This property makes the modified tips highly resistant to degradation during scanning, thus increasing their durability. As demonstrated by the heterogeneous set of structures measured in the present study performed in air, the nanoparticle coated tips are suitable for KPFM analysis. In particular, surface potential difference determination on graphene deposited on silicon, gold sputtered on a salt surface, large and mildly rough areas of ZnO films and small DNA molecules on insulating mica have been achieved with enhanced resolution.

15.
Neurochem Int ; 61(8): 1314-24, 2012 Dec.
Article in English | MEDLINE | ID: mdl-23022607

ABSTRACT

Aquaporin-4 (AQP4) is a water channel protein mainly located in the astroglial plasma membrane, the precise function of which in the brain edema that accompanies hepatic encephalopathy (HE) is unclear. Since ammonia is the main pathogenic agent in HE, its effect on AQP4 expression and distribution in confluent primary astroglial cultures was examined via their exposure to ammonium chloride (1, 3 and 5 mM) for 5 and 10 days. Ammonia induced the general inhibition of AQP4 mRNA synthesis except in the 1 mM/5 day treatment. However, the AQP4 protein content measured was dependent on the method of analysis; an apparent increase was recorded in treated cells in in-cell Western assays, while an apparent reduction was seen with the classic Western blot method, perhaps due to differences in AQP4 aggregation. Ammonia might therefore induce the formation of insoluble AQP4 aggregates in the astroglial plasma membrane. The finding of AQP4 in the pellet of classic Western blot samples, plus data obtained via confocal microscopy, atomic force microscopy (using immunolabeled cells with gold nanoparticles) and scanning electron microscopy, all corroborate this hypothesis. The effect of ammonia on AQP4 seems not to be due to any osmotic effect; identical osmotic stress induced by glutamine and salt had no significant effect on the AQP4 content. AQP4 functional analysis (subjecting astrocytes to a hypo-osmotic medium and using flow cytometry to measure cell size) demonstrated a smaller water influx in ammonia-treated astrocytes suggesting that AQP4 aggregates are representative of an inactive status; however, more confirmatory studies are required to fully understand the functional status of AQP4 aggregates. The present results suggest that ammonia affects AQP4 expression and distribution, and that astrocytes change their expression of AQP4 mRNA as well as the aggregation status of the ensuing protein depending on the ammonia concentration and duration of exposure.


Subject(s)
Ammonia/pharmacology , Aquaporin 4/metabolism , Astrocytes/metabolism , Cell Membrane/chemistry , Membrane Proteins/metabolism , Nerve Tissue Proteins/metabolism , Animals , Aquaporin 4/chemistry , Aquaporin 4/genetics , Astrocytes/ultrastructure , Brain Edema/etiology , Brain Edema/metabolism , Cell Membrane/ultrastructure , Cells, Cultured , Culture Media, Conditioned/chemistry , Flow Cytometry , GRB2 Adaptor Protein/analysis , Glutamine/pharmacology , Hepatic Encephalopathy/complications , Hepatic Encephalopathy/metabolism , Membrane Proteins/chemistry , Membrane Proteins/genetics , Microscopy , Nerve Tissue Proteins/chemistry , Nerve Tissue Proteins/genetics , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Rats , Rats, Wistar , Sodium Chloride/pharmacology , Solubility , Taurine/analysis , alpha-Synuclein/analysis
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