RESUMO
Acoustically excited microstructures have demonstrated significant potential for small-scale biomedical applications by overcoming major microfluidic limitations. Recently, the application of oscillating microbubbles has demonstrated their superiority over acoustically excited solid structures due to their enhanced acoustic streaming at low input power. However, their limited temporal stability hinders their direct applicability for industrial or clinical purposes. Here, we introduce the embedded microbubble, a novel acoustofluidic design based on the combination of solid structures (poly(dimethylsiloxane)) and microbubbles (air-filled cavity) to combine the benefits of both approaches while minimizing their drawbacks. We investigate the influence of various design parameters and geometrical features through numerical simulations and experimentally evaluate their manipulation capabilities. Finally, we demonstrate the capabilities of our design for microfluidic applications by investigating its mixing performance as well as through the controlled rotational manipulation of individual HeLa cells.
Assuntos
Microbolhas , Microfluídica , Acústica , Células HeLa , HumanosRESUMO
Ferromagnetic nanowires are finding use as untethered sensors and actuators for probing micro- and nanoscale biophysical phenomena, such as for localized sensing and application of forces and torques on biological samples, for tissue heating through magnetic hyperthermia, and for microrheology. Quantifying the magnetic properties of individual isolated nanowires is crucial for such applications. Dynamic cantilever magnetometry is used to measure the magnetic properties of individual sub-500 nm diameter polycrystalline nanowires of Ni and Ni80 Co20 fabricated by template-assisted electrochemical deposition. The values are compared with bulk, ensemble measurements when the nanowires are still embedded within their growth matrix. It is found that single-particle and ensemble measurements of nanowires yield significantly different results that reflect inter-nanowire interactions and chemical modifications of the sample during the release process from the growth matrix. The results highlight the importance of performing single-particle characterization for objects that will be used as individual magnetic nanoactuators or nanosensors in biomedical applications.
Assuntos
Magnetometria/métodos , Nanotecnologia/métodos , Nanofios/química , Imãs/químicaRESUMO
Quantitative micromechanical characterization of single cells and multicellular tissues or organisms is of fundamental importance to the study of cellular growth, morphogenesis, and cell-cell interactions. However, due to limited manipulation capabilities at the microscale, systems used for mechanical characterizations struggle to provide complete three-dimensional coverage of individual specimens. Here, we combine an acoustically driven manipulation device with a micro-force sensor to freely rotate biological samples and quantify mechanical properties at multiple regions of interest within a specimen. The versatility of this tool is demonstrated through the analysis of single Lilium longiflorum pollen grains, in combination with numerical simulations, and individual Caenorhabditis elegans nematodes. It reveals local variations in apparent stiffness for single specimens, providing previously inaccessible information and datasets on mechanical properties that serve as the basis for biophysical modelling and allow deeper insights into the biomechanics of these living systems.
Assuntos
Imageamento Tridimensional/métodos , Micromanipulação/instrumentação , Micromanipulação/métodos , Microscopia de Força Atômica/métodos , Análise de Célula Única/instrumentação , Análise de Célula Única/métodos , Acústica , Animais , Fenômenos Biomecânicos , Caenorhabditis elegans/anatomia & histologia , Caenorhabditis elegans/citologia , Parede Celular/ultraestrutura , Lilium/citologia , Microscopia Eletrônica de Varredura , Morfogênese , Células Vegetais , Pólen/citologia , Pólen/ultraestruturaRESUMO
The world was unprepared for the COVID-19 pandemic, and recovery is likely to be a long process. Robots have long been heralded to take on dangerous, dull, and dirty jobs, often in environments that are unsuitable for humans. Could robots be used to fight future pandemics? We review the fundamental requirements for robotics for infectious disease management and outline how robotic technologies can be used in different scenarios, including disease prevention and monitoring, clinical care, laboratory automation, logistics, and maintenance of socioeconomic activities. We also address some of the open challenges for developing advanced robots that are application oriented, reliable, safe, and rapidly deployable when needed. Last, we look at the ethical use of robots and call for globally sustained efforts in order for robots to be ready for future outbreaks.
Assuntos
Controle de Doenças Transmissíveis/tendências , Doenças Transmissíveis , Robótica/tendências , COVID-19/prevenção & controle , Doenças Transmissíveis/diagnóstico , Doenças Transmissíveis/terapia , Desinfecção/tendências , Humanos , Aprendizado de Máquina , Pandemias/prevenção & controle , Tecnologia de Sensoriamento Remoto/tendências , Procedimentos Cirúrgicos Robóticos/tendências , Robótica/instrumentação , SARS-CoV-2 , Interface Usuário-ComputadorRESUMO
Wireless capsule endoscopy (WCE) is a powerful tool for medical screening and diagnosis, where a small capsule is swallowed and moved by means of natural peristalsis and gravity through the human gastrointestinal (GI) tract. The camera-integrated capsule allows for visualization of the small intestine, a region which was previously inaccessible to classical flexible endoscopy. As a diagnostic tool, it allows to localize the sources of bleedings in the middle part of the gastrointestinal tract and to identify diseases, such as inflammatory bowel disease (Crohn's disease), polyposis syndrome, and tumors. The screening and diagnostic efficacy of the WCE, especially in the stomach region, is hampered by a variety of technical challenges like the lack of active capsular position and orientation control. Therapeutic functionality is absent in most commercial capsules, due to constraints in capsular volume and energy storage. The possibility of using body-exogenous magnetic fields to guide, orient, power, and operate the capsule and its mechanisms has led to increasing research in Magnetically Guided Capsule Endoscopy (MGCE). This work shortly reviews the history and state-of-art in WCE technology. It highlights the magnetic technologies for advancing diagnostic and therapeutic functionalities of WCE. Not restricting itself to the GI tract, the review further investigates the technological developments in magnetically guided microrobots that can navigate through the various air- and fluid-filled lumina and cavities in the body for minimally invasive medicine.
Assuntos
Endoscopia por Cápsula , Gastroenteropatias/diagnóstico por imagem , Intestino Delgado/diagnóstico por imagem , Doença de Crohn/diagnóstico por imagem , HumanosRESUMO
An FeGa@P(VDF-TrFE) wire-shaped magnetoelectric nanorobot is designed and fabricated to demonstrate a proof-of-concept integrated device, which features wireless locomotion and on-site triggered therapeutics with a single external power source (i.e., a magnetic field). The device can be precisely steered toward a targeted location wirelessly by rotating magnetic fields and perform on-demand magnetoelectrically assisted drug release to kill cancer cells.
Assuntos
Nanofios , Sistemas de Liberação de Medicamentos , Liberação Controlada de Fármacos , Campos MagnéticosRESUMO
[This corrects the article DOI: 10.1371/journal.pone.0168138.].
RESUMO
Correction for 'High precision, localized proton gradients and fluxes generated by a microelectrode device induce differential growth behaviors of pollen tubes' by Chengzhi Hu et al., Lab Chip, 2017, 17, 671-680.
RESUMO
Pollen tubes are tip-growing plant cells that deliver the sperm cells to the ovules for double fertilization of the egg cell and the endosperm. Various directional cues can trigger the reorientation of pollen tube growth direction on their passage through the female tissues. Among the external stimuli, protons serve an important, regulatory role in the control of pollen tube growth. The generation of local guidance cues has been challenging when investigating the mechanisms of perception and processing of such directional triggers in pollen tubes. Here, we developed and characterized a microelectrode device to generate a local proton gradient and proton flux through water electrolysis. We confirmed that the cytoplasmic pH of pollen tubes varied with environmental pH change. Depending on the position of the pollen tube tip relative to the proton gradient, we observed alterations in the growth behavior, such as bursting at the tip, change in growth direction, or complete growth arrest. Bursting and growth arrest support the hypothesis that changes in the extracellular H+ concentration may interfere with cell wall integrity and actin polymerization at the growing tip. A change in growth direction for some pollen tubes implies that they can perceive the local proton gradient and respond to it. We also showed that the growth rate is directly correlated with the extracellular pH in the tip region. Our microelectrode approach provides a simple method to generate protons and investigate their effect on plant cell growth.
Assuntos
Microeletrodos , Tubo Polínico , Prótons , Técnicas de Cultura de Tecidos/métodos , Desenho de Equipamento , Concentração de Íons de Hidrogênio , Dispositivos Lab-On-A-Chip , Lilium/citologia , Lilium/crescimento & desenvolvimento , Lilium/fisiologia , Tubo Polínico/citologia , Tubo Polínico/crescimento & desenvolvimento , Tubo Polínico/fisiologia , Técnicas de Cultura de Tecidos/instrumentaçãoRESUMO
Fibroblast growth factor 2 (FGF-2), an important paracrine growth factor, binds electrostatically with low micromolar affinity to heparan sulfates present on extracellular matrix proteins. A single molecular analysis served as a basis to decipher the nanomechanical mechanism of the interaction between FGF-2 and the heparan sulfate surrogate, heparin, with a modular atomic force microscope (AFM) design combining magnetic actuators with force measurements at the low force regime (1 × 101 to 1 × 104 pN/s). Unbinding events between FGF-2-heparin complexes were specific and short-lived. Binding between FGF-2 and heparin had strong slip bond characteristics as demonstrated by a decrease of lifetime with tensile force on the complex. Unbinding forces between FGF-2 and heparin were further detailed at different pH as relevant for (patho-) physiological conditions. An acidic pH environment (5.5) modulated FGF-2-heparin binding as demonstrated by enhanced rupture forces needed to release FGF-2 from the heparin-FGF-2 complex as compared to physiological conditions. This study provides a mechanistic and hypothesis driven model on how molecular forces may impact FGF-2 release and storage during tissue remodeling and repair.
RESUMO
The pollen tube is a fast growing cellular protrusion that plays a key role in the reproductive process of flowering plants. It serves as an important model for studying cellular morphogenesis, anisotropic growth mechanisms, and cellular signaling in the plant sciences. The anisotropic growth of pollen tubes is driven by a finely tuned control of the intracellular turgor pressure and the extensibility of the cell wall. To decipher this internal feedback loop and mathematically model the growth process, a quantitative understanding of the mechanical properties of the cell wall is crucial, in addition to biochemical investigations. We report an integrated microfluidic-MEMS force sensor system that allows for high-throughput optical and mechanical investigations of pollen tubes. The system permits large-scale germination, growth, and optical phenotyping of pollen tubes empowering rapid micro-indentation measurements on these cells.
Assuntos
Fenômenos Mecânicos , Sistemas Microeletromecânicos/instrumentação , Células Vegetais , Tubo Polínico/crescimento & desenvolvimento , Fenômenos Biomecânicos , Parede Celular/metabolismo , Modelos Biológicos , Morfogênese , Transdução de SinaisRESUMO
We report a modular atomic force microscope (AFM) design for biomolecular experiments. The AFM head uses readily available components and incorporates deflection-based optics and a piezotube-based cantilever actuator. Jetted-polymers have been used in the mechanical assembly, which allows rapid manufacturing. In addition, a FeCo-tipped electromagnet provides high-force cantilever actuation with vertical magnetic fields up to 0.55 T. Magnetic field calibration has been performed with a micro-hall sensor, which corresponds well with results from finite element magnetostatics simulations. An integrated force resolution of 1.82 and 2.98 pN, in air and in DI water, respectively was achieved in 1 kHz bandwidth with commercially available cantilevers made of Silicon Nitride. The controller and user interface are implemented on modular hardware to ensure scalability. The AFM can be operated in different modes, such as molecular pulling or force-clamp, by actuating the cantilever with the available actuators. The electromagnetic and piezoelectric actuation capabilities have been demonstrated in unbinding experiments of the biotin-streptavidin complex.
Assuntos
Proteínas de Bactérias/ultraestrutura , Biotina/análogos & derivados , Desenho de Equipamento , Microscopia de Força Atômica/instrumentação , Campos Eletromagnéticos , Imãs , Microscopia de Força Atômica/métodosRESUMO
Quantification of mechanical properties of tissues, living cells, and cellular components is crucial for the modeling of plant developmental processes such as mechanotransduction. Pollen tubes are tip-growing cells that provide an ideal system to study the mechanical properties at the single cell level. In this article, a lab-on-a-chip (LOC) device is developed to quantitatively measure the biomechanical properties of lily (Lilium longiflorum) pollen tubes. A single pollen tube is fixed inside the microfluidic chip at a specific orientation and subjected to compression by a soft membrane. By comparing the deformation of the pollen tube at a given external load (compressibility) and the effect of turgor pressure on the tube diameter (stretch ratio) with finite element modeling, its mechanical properties are determined. The turgor pressure and wall stiffness of the pollen tubes are found to decrease considerably with increasing initial diameter of the pollen tubes. This observation supports the hypothesis that tip-growth is regulated by a delicate balance between turgor pressure and wall stiffness. The LOC device is modular and adaptable to a variety of cells that exhibit tip-growth, allowing for the straightforward measurement of mechanical properties.
Assuntos
Dispositivos Lab-On-A-Chip , Tubo Polínico/crescimento & desenvolvimento , Módulo de Elasticidade , Desenho de Equipamento , Lilium/crescimento & desenvolvimento , Lilium/metabolismo , Microfluídica/instrumentação , Microfluídica/métodos , Microscopia Eletrônica , Tubo Polínico/químicaRESUMO
Pollen tubes are used as a model in the study of plant morphogenesis, cellular differentiation, cell wall biochemistry, biomechanics, and intra- and intercellular signaling. For a "systems-understanding" of the bio-chemo-mechanics of tip-polarized growth in pollen tubes, the need for a versatile, experimental assay platform for quantitative data collection and analysis is critical. We introduce a Lab-on-a-Chip (LoC) concept for high-throughput pollen germination and pollen tube guidance for parallelized optical and mechanical measurements. The LoC localizes a large number of growing pollen tubes on a single plane of focus with unidirectional tip-growth, enabling high-resolution quantitative microscopy. This species-independent LoC platform can be integrated with micro-/nano-indentation systems, such as the cellular force microscope (CFM) or the atomic force microscope (AFM), allowing for rapid measurements of cell wall stiffness of growing tubes. As a demonstrative example, we show the growth and directional guidance of hundreds of lily (Lilium longiflorum) and Arabidopsis (Arabidopsis thaliana) pollen tubes on a single LoC microscopy slide. Combining the LoC with the CFM, we characterized the cell wall stiffness of lily pollen tubes. Using the stiffness statistics and finite-element-method (FEM)-based approaches, we computed an effective range of the linear elastic moduli of the cell wall spanning the variability space of physiological parameters including internal turgor, cell wall thickness, and tube diameter. We propose the LoC device as a versatile and high-throughput phenomics platform for plant reproductive and development biology using the pollen tube as a model.
Assuntos
Parede Celular/fisiologia , Dispositivos Lab-On-A-Chip , Lilium/fisiologia , Tubo Polínico/crescimento & desenvolvimento , Módulo de Elasticidade , Polinização/fisiologiaRESUMO
We report a novel atomic force microscopy (AFM) technique with dual actuation capabilities using both piezo and magnetic bead actuation for advanced single-molecule force spectroscopy experiments. The experiments are performed by manipulating magnetic microbeads using an electromagnet against a stationary cantilever. Magnetic actuation has been demonstrated before to actuate cantilevers, but here we keep the cantilever stationary and accomplish actuation via free-manipulated microstructures. The developed method benefits from significant reduction of drift, since the experiments are performed without a substrate contact and the measured force is inherently differential. In addition, shrinking the size of the actuator can minimize hydrodynamic forces affecting the cantilever. The new method reported herein allows for the application of constant force to perform force-clamp experiments without any active feedback, profiled for a deeper understanding of biomolecular interactions.
RESUMO
In this paper, we report on the synthesis of FeCo/Cu multisegmented nanowires by means of pulse electrodeposition in nanoporous anodic aluminum oxide arrays supported on silicon chips. By adjustment of the electrodeposition conditions, such as the pulse scheme and the electrolyte, alternating segments of Cu and ferromagnetic FeCo alloy can be fabricated. The segments can be built with a wide range of lengths (15-150 nm) and exhibit a close-to-pure composition (Cu or FeCo alloy) as suggested by energy-dispersive X-ray mapping results. The morphology and the crystallographic structure of different nanowire configurations have been assessed thoroughly, concluding that Fe, Co, and Cu form solid solution. Magnetic characterization using vibrating sample magnetometry and magnetic force microscopy reveals that by introduction of nonmagnetic Cu segments within the nanowire architecture, the magnetic easy axis can be modified and the reduced remanence can be tuned to the desired values. The experimental results are in agreement with the provided simulations. Furthermore, the influence of nanowire magnetic architecture on the magnetically triggered protein desorption is evaluated for three types of nanowires: Cu, FeCo, and multisegmented FeCo15nm/Cu15nm. The application of an external magnetic field can be used to enhance the release of proteins on demand. For fully magnetic FeCo nanowires the applied oscillating field increased protein release by 83%, whereas this was found to be 45% for multisegmented FeCo15nm/Cu15nm nanowires. Our work suggests that a combination of arrays of nanowires with different magnetic configurations could be used to generate complex substance concentration gradients or control delivery of multiple drugs and macromolecules.
Assuntos
Nanopartículas Metálicas/química , Nanopartículas Metálicas/efeitos da radiação , Nanofios/química , Nanofios/efeitos da radiação , Proteínas/química , Absorção Fisico-Química/efeitos da radiação , Adsorção/efeitos da radiação , Cobalto/química , Cobre/química , Cristalização/métodos , Galvanoplastia/métodos , Ferro/química , Campos Magnéticos , Teste de Materiais , Nanopartículas Metálicas/ultraestrutura , Nanofios/ultraestrutura , Ligação Proteica/efeitos da radiação , Proteínas/efeitos da radiaçãoRESUMO
We report the design, fabrication, and characterization of cantilevers with integrated AlN actuators and conductive PtSi tips for multi-frequency atomic force microscopy. These cantilevers also possess a stepped-rectangular geometry. The excellent dynamic behavior of these cantilevers is investigated using both finite-element simulations and experimental methods. Several imaging experiments are presented to illustrate the efficacy and versatility of these cantilevers.