Cell membrane conformation at vertical nanowire array interface revealed by fluorescence imaging

Berthing, Trine; Bonde, Sara; Rostgaard, Katrine R.; Madsen, Morten Hannibal; Sørensen, C. B.; Nygård, Jesper; Martinez, Karen L.

doi:10.1088/0957-4484/23/41/415102

Cited by 98 publications

(143 citation statements)

References 22 publications

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“…The cells were then incubated with 5 μM fluorescent SNAP label (SNAP-Surface 549) for 5 minutes before being washed and fixed. The HEK/SNAP staining technique has previously been used to establish membrane continuity around nanofeatures17. All cells were imaged in PBS with confocal microscopy to isolate the basal membrane.…”

Section: Resultsmentioning

confidence: 99%

Biomimetic surface patterning for long-term transmembrane access

VanDersarl

Renaud

2016

Sci Rep

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Here we present a planar patch clamp chip based on biomimetic cell membrane fusion. This architecture uses nanometer length-scale surface patterning to replicate the structure and function of membrane proteins, creating a gigaohm seal between the cell and a planar electrode array. The seal is generated passively during cell spreading, without the application of a vacuum to the cell surface. This interface can enable cell-attached and whole-cell recordings that are stable to 72 hours, and generates no visible damage to the cell. The electrodes can be very small (<5 μm) and closely packed, offering a high density platform for cellular measurement.

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Section: Resultsmentioning

confidence: 99%

Biomimetic surface patterning for long-term transmembrane access

VanDersarl

Renaud

2016

Sci Rep

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“…27−31 The PC-12 cells cultured on the VNEA were further characterized by z-step fluorescent scanning with two-photon microscopy ( Figure 2d). 19,32,33 The cross-sectional images (Figure 2e) clearly show that the cells invaginate the NW electrodes. We also observed several vertical NW electrode−cell interfaces by SEM and confirmed that the electrodes penetrate and locate inside of cytoplasm of the cells by the cross-sectional SEM image (Supporting Information, Figure S2).…”

Section: T He Ephaptic Interaction Between Living Cells and Electricalmentioning

confidence: 97%

Enhanced Neurite Outgrowth by Intracellular Stimulation

Kim¹,

Lee

Kim³

et al. 2015

Nano Lett.

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Electrical stimulation through direct electrical activation has been widely used to recover the function of neurons, primarily through the extracellular application of thin film electrodes. However, studies using extracellular methods show limited ability to reveal correlations between the cells and the electrical stimulation due to interference from external sources such as membrane capacitance and culture medium. Here, we demonstrate long-term intracellular electrical stimulation of undamaged pheochromocytoma (PC-12) cells by utilizing a vertical nanowire electrode array (VNEA). The VNEA was prepared by synthesizing silicon nanowires on a Si substrate through a vapor-liquid-solid (VLS) mechanism and then fabricating them into electrodes with semiconductor nanodevice processing. PC-12 cells were cultured on the VNEA for 4 days with intracellular electrical stimulation and then a 2-day stabilization period. Periodic scanning via two-photon microscopy confirmed that the electrodes pierced the cells without inducing damage. Electrical stimulation through the VNEA enhances cellular differentiation and neurite outgrowth by about 50% relative to extracellular stimulation under the same conditions. VNEA-mediated stimulation also revealed that cellular differentiation and growth in the cultures were dependent on the potential used to stimulate them. Intracellular stimulation using nanowires could pave the way for controlled cellular differentiation and outgrowth studies in living cells.

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“…Due to the high aspect ratio and the characteristic nanoscale dimensions, nanoneedles intimately contact large traits of the cell membrane, strongly interacting with it and with the intracellular milieu 16,37,38 ( Figure 2). It is this unique interaction, whose details are still largely unexplored, that enables their superior ability to investigate the intracellular space.…”

Section: The Biointerfacementioning

confidence: 99%

Nanoneedle-Based Sensing in Biological Systems

Chiappini

2017

ACS Sens.

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Nanoneedles are high aspect ratio nanostructures with a unique biointerface. Thanks to their peculiar yet poorly understood interaction with cells, they very effectively sense intracellular conditions, typically with lower toxicity and perturbation than traditionally available probes. Through long-term, reversible interfacing with cells, nanoneedles can monitor biological functions over the course of several days. Their nanoscale dimension and the assembly into large scale, ordered, dense arrays enable monitoring the functions of large cell populations, to provide functional maps with sub-micron spatial resolution. Intracellularly, they sense electrical activity of complex excitable networks, as well as concentration, function and interaction of biomolecules in situ, while extracellularly they can measure the forces exerted by cells with piconewton detection limits, or efficiently sort rare cells based on their membrane receptors. Nanoneedles can investigate the function of many biological systems, ranging from cells, to biological fluids, to tissues and living organisms. This review examines the devices, strategies and workflows developed to use nanoneedles for sensing in biological systems. KeywordsNanoneedles, nanowires, biointerface, intracellular sensing, review, biomarkers, label-free, minimally invasive.Nanoneedles are rapidly emerging as a tool to interact with the intracellular environment of large number of cells simultaneously, with limited perturbation of their physiological processes. This interaction provides characteristics advantages for minimally invasive cell and molecular biology investigations, as well as to progress biomedical translation of regenerative and precision medicine approaches. A quick string of several successful proofs of principles have established nanoneedles' potential to efficiently deliver impermeant molecules 1,2 and nanoparticles 3 directly to the cell cytosol, and to sense the intracellular milieu 4-6 across biological systems ranging from cells in culture 7 to living organisms 8 . Nanoneedles can be broadly defined as nanomaterials whose high aspect ratio enables their biological interaction; this definition encompasses a broad range of classically defined nanomaterials, which cater for different biointerfacing needs and applications ( Figure 1). Figure 1 Overview of the nanoneedle devices and strategies used for sensing in biological systems.Each nanoneedle is a stacked bar of a bar graph. The legend below each nanoneedle is color-coded and connects to the associated bar. The height of each bar represents the fraction of reviewed papers with that characteristic. The number in each bar is the number of reviewed publications with that characteristic. Numbers for each bar do not sum to the same total as publications can contribute to more than one bar in a given stack. *"Fluid" excludes biological fluids, + "in vivo/ex vivo" includes biological fluids. The overview is compiled from all the publications reviewed herein that include sensing work.Nanoneedles belong t...

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Cell membrane conformation at vertical nanowire array interface revealed by fluorescence imaging

Cited by 98 publications

References 22 publications

Biomimetic surface patterning for long-term transmembrane access

Biomimetic surface patterning for long-term transmembrane access

Enhanced Neurite Outgrowth by Intracellular Stimulation

Nanoneedle-Based Sensing in Biological Systems

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