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1.
Nat Methods ; 19(6): 751-758, 2022 06.
Artigo em Inglês | MEDLINE | ID: mdl-35637303

RESUMO

Label-free characterization of single biomolecules aims to complement fluorescence microscopy in situations where labeling compromises data interpretation, is technically challenging or even impossible. However, existing methods require the investigated species to bind to a surface to be visible, thereby leaving a large fraction of analytes undetected. Here, we present nanofluidic scattering microscopy (NSM), which overcomes these limitations by enabling label-free, real-time imaging of single biomolecules diffusing inside a nanofluidic channel. NSM facilitates accurate determination of molecular weight from the measured optical contrast and of the hydrodynamic radius from the measured diffusivity, from which information about the conformational state can be inferred. Furthermore, we demonstrate its applicability to the analysis of a complex biofluid, using conditioned cell culture medium containing extracellular vesicles as an example. We foresee the application of NSM to monitor conformational changes, aggregation and interactions of single biomolecules, and to analyze single-cell secretomes.


Assuntos
Nanopartículas , Nanotecnologia , Difusão , Microscopia de Fluorescência
2.
Nano Lett ; 24(6): 1874-1881, 2024 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-38295760

RESUMO

Traditional single-nanoparticle sizing using optical microscopy techniques assesses size via the diffusion constant, which requires suspended particles to be in a medium of known viscosity. However, these assumptions are typically not fulfilled in complex natural sample environments. Here, we introduce dual-angle interferometric scattering microscopy (DAISY), enabling optical quantification of both size and polarizability of individual nanoparticles (radius <170 nm) without requiring a priori information regarding the surrounding media or super-resolution imaging. DAISY achieves this by combining the information contained in concurrently measured forward and backward scattering images through twilight off-axis holography and interferometric scattering (iSCAT). Going beyond particle size and polarizability, single-particle morphology can be deduced from the fact that the hydrodynamic radius relates to the outer particle radius, while the scattering-based size estimate depends on the internal mass distribution of the particles. We demonstrate this by differentiating biomolecular fractal aggregates from spherical particles in fetal bovine serum at the single-particle level.

3.
Nano Lett ; 21(19): 8503-8509, 2021 10 13.
Artigo em Inglês | MEDLINE | ID: mdl-34403260

RESUMO

During diffusion of nanoparticles bound to a cellular membrane by ligand-receptor pairs, the distance to the laterally mobile interface is sufficiently short for their motion to depend not only on the membrane-mediated diffusivity of the tethers but also in a not yet fully understood manner on nanoparticle size and interfacial hydrodynamics. By quantifying diffusivity, velocity, and size of individual membrane-bound liposomes subjected to a hydrodynamic shear flow, we have successfully separated the diffusivity contributions from particle size and number of tethers. The obtained diffusion-size relations for synthetic and extracellular lipid vesicles are not well-described by the conventional no-slip boundary condition, suggesting partial slip as well as a significant diffusivity dependence on the distance to the lipid bilayer. These insights, extending the understanding of diffusion of biological nanoparticles at lipid bilayers, are of relevance for processes such as cellular uptake of viruses and lipid nanoparticles or labeling of cell-membrane-residing molecules.


Assuntos
Bicamadas Lipídicas , Lipossomos , Membrana Celular , Difusão , Membranas
4.
Anal Chem ; 92(2): 1908-1915, 2020 01 21.
Artigo em Inglês | MEDLINE | ID: mdl-31820950

RESUMO

Determination of size and refractive index (RI) of dispersed unlabeled subwavelength particles is of growing interest in several fields, including biotechnology, wastewater monitoring, and nanobubble preparations. Conventionally, the size distribution of such samples is determined via the Brownian motion of the particles, but simultaneous determination of their RI remains challenging. This work demonstrates nanoparticle tracking analysis (NTA) in an off-axis digital holographic microscope (DHM) enabling determination of both particle size and RI of individual subwavelength particles from the combined information about size and optical phase shift. The potential of the method to separate particle populations is demonstrated by analyzing a mixture of three types of dielectric particles within a narrow size range, where conventional NTA methods based on Brownian motion alone would fail. Using this approach, the phase shift allowed individual populations of dielectric beads overlapping in either size or RI to be clearly distinguished and quantified with respect to these properties. The method was furthermore applied for analysis of surfactant-stabilized micro- and nanobubbles, with RI lower than that of water. Since bubbles induce a phase shift of opposite sign to that of solid particles, they were easily distinguished from similarly sized solid particles made up of undissolved surfactant. Surprisingly, the dependence of the phase shift on bubble size indicates that only those with 0.15-0.20 µm radius were individual bubbles, whereas larger bubbles were actually clusters of bubbles. This label-free means to quantify multiple parameters of suspended individual submicrometer particles offers a crucial complement to current characterization strategies, suggesting broad applicability for a wide range of nanoparticle systems.


Assuntos
Ar , Nanopartículas/química , Tamanho da Partícula , Poliestirenos/química , Refratometria , Dióxido de Silício/química , Hexoses/química , Microbolhas , Polissorbatos/química , Tensoativos/química
6.
Nanotechnology ; 27(23): 238001, 2016 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-27121075

RESUMO

We compare the simplified valence-force model for single-layer black phosphorus with the original model and recent ab initio results. Using an analytic approach and numerical calculations we find that the simplified model yields Young's moduli that are smaller compared to the original model and are almost a factor of two smaller than ab initio results. Moreover, the Poisson ratios are an order of magnitude smaller than values found in the literature.

7.
Phys Chem Chem Phys ; 18(33): 23312-9, 2016 Aug 17.
Artigo em Inglês | MEDLINE | ID: mdl-27499171

RESUMO

In order to understand the relation of strain and material properties, both a microscopic model connecting a given strain to the displacement of atoms, and a macroscopic model relating applied stress to induced strain, are required. Starting from a valence-force model for black phosphorous [Kaneta et al., Solid State Communications, 1982, 44, 613] we use recent experimental and computational results to obtain an improved set of valence-force parameters for phosphorene. From the model we calculate the phonon dispersion and the elastic properties of single-layer phosphorene. Finally, we use these results to derive a complete continuum model, including the bending rigidities, valid for long-wavelength deformations of phosphorene. This continuum model is then used to study the properties of pressurized suspended phosphorene sheets.

8.
Phys Rev Lett ; 112(14): 145503, 2014 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-24765986

RESUMO

Thermalization in nonlinear systems is a central concept in statistical mechanics and has been extensively studied theoretically since the seminal work of Fermi, Pasta, and Ulam. Using molecular dynamics and continuum modeling of a ring-down setup, we show that thermalization due to nonlinear mode coupling intrinsically limits the quality factor of nanomechanical graphene drums and turns them into potential test beds for Fermi-Pasta-Ulam physics. We find the thermalization rate Γ to be independent of radius and scaling as Γ∼T*/εpre2, where T* and εpre are effective resonator temperature and prestrain.

9.
Nat Commun ; 15(1): 1208, 2024 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-38332035

RESUMO

Environmental humidity variations are ubiquitous and high humidity characterizes fuel cell and electrolyzer operation conditions. Since hydrogen-air mixtures are highly flammable, humidity tolerant H2 sensors are important from safety and process monitoring perspectives. Here, we report an optical nanoplasmonic hydrogen sensor operated at elevated temperature that combined with Deep Dense Neural Network or Transformer data treatment involving the entire spectral response of the sensor enables a 100 ppm H2 limit of detection in synthetic air at 80% relative humidity. This significantly exceeds the <1000 ppm US Department of Energy performance target. Furthermore, the sensors pass the ISO 26142:2010 stability requirement in 80% relative humidity in air down to 0.06% H2 and show no signs of performance loss after 140 h continuous operation. Our results thus demonstrate the potential of plasmonic hydrogen sensors for use in high humidity and how neural-network-based data treatment can significantly boost their performance.

10.
Nano Lett ; 12(7): 3526-31, 2012 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-22708530

RESUMO

Classical continuum mechanics is used extensively to predict the properties of nanoscale materials such as graphene. The bending rigidity, κ, is an important parameter that is used, for example, to predict the performance of graphene nanoelectromechanical devices and also ripple formation. Despite its importance, there is a large spread in the theoretical predictions of κ for few-layer graphene. We have used the snap-through behavior of convex buckled graphene membranes under the application of electrostatic pressure to determine experimentally values of κ for double-layer graphene membranes. We demonstrate how to prepare convex-buckled suspended graphene ribbons and fully clamped suspended membranes and show how the determination of the curvature of the membranes and the critical snap-through voltage, using AFM, allows us to extract κ. The bending rigidity of bilayer graphene membranes under ambient conditions was determined to be 35.5−15.0 +20.0 eV. Monolayers are shown to have significantly lower κ than bilayers.

11.
ArXiv ; 2023 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-36945686

RESUMO

Through digital imaging, microscopy has evolved from primarily being a means for visual observation of life at the micro- and nano-scale, to a quantitative tool with ever-increasing resolution and throughput. Artificial intelligence, deep neural networks, and machine learning are all niche terms describing computational methods that have gained a pivotal role in microscopy-based research over the past decade. This Roadmap is written collectively by prominent researchers and encompasses selected aspects of how machine learning is applied to microscopy image data, with the aim of gaining scientific knowledge by improved image quality, automated detection, segmentation, classification and tracking of objects, and efficient merging of information from multiple imaging modalities. We aim to give the reader an overview of the key developments and an understanding of possibilities and limitations of machine learning for microscopy. It will be of interest to a wide cross-disciplinary audience in the physical sciences and life sciences.

12.
Nano Lett ; 11(4): 1439-42, 2011 Apr 13.
Artigo em Inglês | MEDLINE | ID: mdl-21375279

RESUMO

We show that the coupling between single-electron charging and mechanical motion in a nanoelectromechanical single-electron transistor can be utilized in a novel parametric actuation scheme. This scheme, which relies on a periodic modulation of the mechanical resonance frequency through an alternating source-drain voltage, leads to a parametric instability and emergence of mechanical vibrations in a limited range of modulation amplitudes. Remarkably, the frequency range where instability occurs and the maximum oscillation amplitude, depend weakly on the damping in the system. We also show that a weak parametric modulation increases the effective quality factor and amplifies the system's response to the conventional actuation that exploits an AC gate signal.


Assuntos
Sistemas Microeletromecânicos/instrumentação , Transistores Eletrônicos , Desenho Assistido por Computador , Elétrons , Desenho de Equipamento , Análise de Falha de Equipamento
13.
Nano Lett ; 11(9): 3569-75, 2011 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-21848317

RESUMO

Novel field effect transistors with suspended graphene gates are demonstrated. By incorporating mechanical motion of the gate electrode, it is possible to improve the switching characteristics compared to a static gate, as shown by a combination of experimental measurements and numerical simulations. The mechanical motion of the graphene gate is confirmed by using atomic force microscopy to directly measure the electrostatic deflection. The device geometry investigated here can also provide a sensitive measurement technique for detecting high-frequency motion of suspended membranes as required, e.g., for mass sensing.


Assuntos
Grafite/química , Nanotecnologia/métodos , Nanotubos de Carbono/química , Condutividade Elétrica , Eletrodos , Microscopia de Força Atômica/métodos , Eletricidade Estática , Temperatura
14.
Elife ; 112022 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-36317499

RESUMO

The marine microbial food web plays a central role in the global carbon cycle. However, our mechanistic understanding of the ocean is biased toward its larger constituents, while rates and biomass fluxes in the microbial food web are mainly inferred from indirect measurements and ensemble averages. Yet, resolution at the level of the individual microplankton is required to advance our understanding of the microbial food web. Here, we demonstrate that, by combining holographic microscopy with deep learning, we can follow microplanktons throughout their lifespan, continuously measuring their three-dimensional position and dry mass. The deep-learning algorithms circumvent the computationally intensive processing of holographic data and allow rapid measurements over extended time periods. This permits us to reliably estimate growth rates, both in terms of dry mass increase and cell divisions, as well as to measure trophic interactions between species such as predation events. The individual resolution provides information about selectivity, individual feeding rates, and handling times for individual microplanktons. The method is particularly useful to detail the rates and routes of organic matter transfer in micro-zooplankton, the most important and least known group of primary consumers in the oceans. Studying individual interactions in idealized small systems provides insights that help us understand microbial food webs and ultimately larger-scale processes. We exemplify this by detailed descriptions of micro-zooplankton feeding events, cell divisions, and long-term monitoring of single cells from division to division.


Picture a glass of seawater. It looks clear and empty, but in reality, it contains one hundred million bacteria, about one hundred thousand other single-celled organisms, and a few microscopic animals. In fact, the majority of life in the ocean is microscopic and we know relatively little about it. Nevertheless, these microbes have a major impact on our lives. Microscopic algae known as phytoplankton, for example, produce half of the oxygen we breathe. For animals, birds and other large organisms in the ocean, we have a good understanding of who eats who and where the material ends up. However, for phytoplankton and other microbes, we depend on bulk measurements and averages of large groups. Bachimanchi et al. developed a method to follow individual microbes living in seawater and to observe how they move, grow, consume each other and reproduce. The team combined holographic microscopy with artificial intelligence to follow multiple planktons, diatoms and other microbes throughout their life span and continuously measured their three-dimensional location and mass. This made it possible to estimate how fast the organisms were growing and moving, and to observe what they ate. The experiments revealed new insights into how micro-zooplankton, diatoms and other microbes in the ocean interact with each other. This new method may be useful for researchers who would like to track the movements and whereabouts of microscopic planktons, bacteria or other microbes for extended periods of time. It is also a rapid method for counting, sizing, and weighing cells in suspension. The hardware used in this method is relatively cheap, and Bachimanchi et al. have shared all the computer code with examples and demonstrations in a public database to enable other researchers to use it.


Assuntos
Aprendizado Profundo , Fitoplâncton , Animais , Microscopia , Zooplâncton , Oceanos e Mares , Água do Mar
15.
Nat Commun ; 13(1): 7492, 2022 12 05.
Artigo em Inglês | MEDLINE | ID: mdl-36470883

RESUMO

Object detection is a fundamental task in digital microscopy, where machine learning has made great strides in overcoming the limitations of classical approaches. The training of state-of-the-art machine-learning methods almost universally relies on vast amounts of labeled experimental data or the ability to numerically simulate realistic datasets. However, experimental data are often challenging to label and cannot be easily reproduced numerically. Here, we propose a deep-learning method, named LodeSTAR (Localization and detection from Symmetries, Translations And Rotations), that learns to detect microscopic objects with sub-pixel accuracy from a single unlabeled experimental image by exploiting the inherent roto-translational symmetries of this task. We demonstrate that LodeSTAR outperforms traditional methods in terms of accuracy, also when analyzing challenging experimental data containing densely packed cells or noisy backgrounds. Furthermore, by exploiting additional symmetries we show that LodeSTAR can measure other properties, e.g., vertical position and polarizability in holographic microscopy.


Assuntos
Holografia , Microscopia , Algoritmos , Aprendizado de Máquina
16.
ACS Nano ; 15(2): 2240-2250, 2021 02 23.
Artigo em Inglês | MEDLINE | ID: mdl-33399450

RESUMO

Characterization of suspended nanoparticles in their native environment plays a central role in a wide range of fields, from medical diagnostics and nanoparticle-enhanced drug delivery to nanosafety and environmental nanopollution assessment. Standard optical approaches for nanoparticle sizing assess the size via the diffusion constant and, as a consequence, require long trajectories and that the medium has a known and uniform viscosity. However, in most biological applications, only short trajectories are available, while simultaneously, the medium viscosity is unknown and tends to display spatiotemporal variations. In this work, we demonstrate a label-free method to quantify not only size but also refractive index of individual subwavelength particles using 2 orders of magnitude shorter trajectories than required by standard methods and without prior knowledge about the physicochemical properties of the medium. We achieved this by developing a weighted average convolutional neural network to analyze holographic images of single particles, which was successfully applied to distinguish and quantify both size and refractive index of subwavelength silica and polystyrene particles without prior knowledge of solute viscosity or refractive index. We further demonstrate how these features make it possible to temporally resolve aggregation dynamics of 31 nm polystyrene nanoparticles, revealing previously unobserved time-resolved dynamics of the monomer number and fractal dimension of individual subwavelength aggregates.

17.
Biophys Rev (Melville) ; 2(3): 031401, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-38505631

RESUMO

Quantitative analysis of cell structures is essential for biomedical and pharmaceutical research. The standard imaging approach relies on fluorescence microscopy, where cell structures of interest are labeled by chemical staining techniques. However, these techniques are often invasive and sometimes even toxic to the cells, in addition to being time consuming, labor intensive, and expensive. Here, we introduce an alternative deep-learning-powered approach based on the analysis of bright-field images by a conditional generative adversarial neural network (cGAN). We show that this is a robust and fast-converging approach to generate virtually stained images from the bright-field images and, in subsequent downstream analyses, to quantify the properties of cell structures. Specifically, we train a cGAN to virtually stain lipid droplets, cytoplasm, and nuclei using bright-field images of human stem-cell-derived fat cells (adipocytes), which are of particular interest for nanomedicine and vaccine development. Subsequently, we use these virtually stained images to extract quantitative measures about these cell structures. Generating virtually stained fluorescence images is less invasive, less expensive, and more reproducible than standard chemical staining; furthermore, it frees up the fluorescence microscopy channels for other analytical probes, thus increasing the amount of information that can be extracted from each cell. To make this deep-learning-powered approach readily available for other users, we provide a Python software package, which can be easily personalized and optimized for specific virtual-staining and cell-profiling applications.

18.
Nat Commun ; 10(1): 340, 2019 01 21.
Artigo em Inglês | MEDLINE | ID: mdl-30664642

RESUMO

Microorganisms adapt their biophysical properties in response to changes in their local environment. However, quantifying these changes at the single-cell level has only recently become possible, largely relying on fluorescent labeling strategies. In this work, we utilize yeast (Saccharomyces cerevisiae) to demonstrate label-free quantification of changes in both intracellular osmolarity and macromolecular concentration in response to changes in the local environment. By combining a digital holographic microscope with a millifluidic chip, the temporal response of cellular water flux was successfully isolated from the rate of production of higher molecular weight compounds, in addition to identifying the produced compounds in terms of the product of their refractive index increment [Formula: see text] and molar mass. The ability to identify, quantify and temporally resolve multiple biophysical processes in living cells at the single cell level offers a crucial complement to label-based strategies, suggesting broad applicability in studies of a wide-range of cellular processes.


Assuntos
Citosol/metabolismo , Saccharomyces cerevisiae/química , Análise de Célula Única/métodos , Água/metabolismo , Transporte Biológico , Citosol/química , Citosol/ultraestrutura , Holografia , Dispositivos Lab-On-A-Chip , Concentração Osmolar , Pressão Osmótica , Refratometria , Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/ultraestrutura , Análise de Célula Única/instrumentação , Água/química
19.
J Phys Chem Lett ; 9(9): 2278-2284, 2018 May 03.
Artigo em Inglês | MEDLINE | ID: mdl-29624391

RESUMO

Quartz crystal microbalance with dissipation monitoring and total internal reflection fluorescence microscopy have been used to investigate binding of norovirus-like particles (noroVLPs) to a supported (phospho)lipid bilayer (SLB) containing a few percent of H or B type 1 glycosphingolipid (GSL) receptors. Although neither of these GSLs spontaneously form domains, noroVLPs were observed to form micron-sized clusters containing typically up to about 30 VLP copies, especially for B type 1, which is a higher-affinity receptor. This novel finding is explained by proposing a model implying that VLP-induced membrane deformation promotes VLP clustering, a hypothesis that was further supported by observing that functionalized gold nanoparticles were able to locally induce SLB deformation. Because similar effects are likely possible also at cellular membranes, our findings are interesting beyond a pure biophysicochemical perspective as they shed new light on what may happen during receptor-mediated uptake of viruses as well as nanocarriers in drug delivery.


Assuntos
Glicoesfingolipídeos/química , Bicamadas Lipídicas/metabolismo , Nanopartículas Metálicas/química , Norovirus/química , Carbocianinas/química , Fluorescência , Corantes Fluorescentes/química , Ouro/química , Humanos , Bicamadas Lipídicas/química , Microscopia de Fluorescência , Fosfatidilcolinas/química , Fosfatidilcolinas/metabolismo
20.
J Phys Condens Matter ; 29(18): 185702, 2017 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-28294103

RESUMO

A multi-scale approach for the theoretical description of deformed phosphorene is presented. This approach combines a valence-force model to relate macroscopic strain to microscopic displacements of atoms and a tight-binding model with distance-dependent hopping parameters to obtain electronic properties. The resulting self-consistent electromechanical model is suitable for large-scale modeling of phosphorene devices. We demonstrate this for the case of inhomogeneously deformed phosphorene drum, which may be used as an exciton funnel.

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