Multifunctional carbon-nanotube cellular endoscopes

Singhal, Riju; Orynbayeva, Zulfiya; Sundaram, Ramalingam Venkat Kalyana; Niu, Jun; Bhattacharyya, Sayan; Vitol, Elina A.; Schrlau, Michael G.; Papazoglou, Elisabeth S.; Friedman, Gary D.; Gogotsi, Yury

doi:10.1038/nnano.2010.241

Cited by 227 publications

(226 citation statements)

References 33 publications

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“…Due to their high-aspect ratio geometry and exceptionally sharp tips, nanopipettes offer an important advantage over conventional nanopores as they can be easily adapted for use in single cell interrogation 15,16 and as a macroscopic delivery vehicle for intracellular injection. [17][18][19][20][21][22] . To date, delivery of molecules has so far only been quantified in close proximity to the tip by using fluorescence spectroscopy.…”

Section: Introductionmentioning

confidence: 99%

On-Demand Delivery of Single DNA Molecules Using Nanopipets

et al. 2015

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Understanding the behavioral properties of single molecules or larger scale populations interacting with single molecules is currently a hotly pursued topic in nanotechnology. This arises from the potential such techniques have in relation to applications such as targeted drug delivery, early stage detection of disease, and drug screening. Although label and label-free single molecule detection strategies have existed for a number of years, currently lacking are efficient methods for the controllable delivery of single molecules in aqueous environments. In this article we show both experimentally and from simulations that nanopipets in conjunction with asymmetric voltage pulses can be used for label-free detection and delivery of single molecules through the tip of a nanopipet with "on-demand" timing resolution. This was demonstrated by controllable delivery of 5 kbp and 10 kbp DNA molecules from solutions with concentrations as low as 3 pM.

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Section: Introductionmentioning

confidence: 99%

On-Demand Delivery of Single DNA Molecules Using Nanopipets

et al. 2015

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“…Several groups have demonstrated the insertion of high aspectratio nanostructures for molecular delivery with minimal cell perturbation, suggesting that nanomaterials may pass through the cell membrane without dire consequences (19)(20)(21)(22)(23)(24). Extending this approach, Na et al (25) showed a nanowire "sandwich assay" in which nanowires approximately 100 nm in diameter functionalized with antibodies penetrate the cell with limited toxicity and can bind specific enzymes for extraction.…”

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confidence: 99%

Nondestructive nanostraw intracellular sampling for longitudinal cell monitoring

Cao

Hjort

Chen

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

131

210

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Here, we report a method for time-resolved, longitudinal extraction and quantitative measurement of intracellular proteins and mRNA from a variety of cell types. Cytosolic contents were repeatedly sampled from the same cell or population of cells for more than 5 d through a cell-culture substrate, incorporating hollow 150-nmdiameter nanostraws (NS) within a defined sampling region. Once extracted, the cellular contents were analyzed with conventional methods, including fluorescence, enzymatic assays (ELISA), and quantitative real-time PCR. This process was nondestructive with >95% cell viability after sampling, enabling long-term analysis. It is important to note that the measured quantities from the cell extract were found to constitute a statistically significant representation of the actual contents within the cells. Of 48 mRNA sequences analyzed from a population of cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs), 41 were accurately quantified. The NS platform samples from a select subpopulation of cells within a larger culture, allowing native cell-to-cell contact and communication even during vigorous activity such as cardiomyocyte beating. This platform was applied both to cell lines and to primary cells, including CHO cells, hiPSC-CMs, and human astrocytes derived in 3D cortical spheroids. By tracking the same cell or group of cells over time, this method offers an avenue to understand dynamic cell behavior, including processes such as induced pluripotency and differentiation.sampling | nanotechnology | molecular biology | cellular biology

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“…Researchers must investigate the reactivity of a protein with its physiological partners to understand its role in cellular pathways; concentration measurements alone could be misleading, because the local environment of a protein can drastically alter its activity. Emerging capillary micro-/nanofabrication technologies allow the chemical reactivity of the target species to be investigated (3,(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21). Allbritton and coworkers (11,12) embarked on the development of this technology and used capillary electrophoresis to analyze intracellular kinase activity.…”

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confidence: 99%

Nanokit for single-cell electrochemical analyses

Pan

Jiang

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

115

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The development of more intricate devices for the analysis of small molecules and protein activity in single cells would advance our knowledge of cellular heterogeneity and signaling cascades. Therefore, in this study, a nanokit was produced by filling a nanometersized capillary with a ring electrode at the tip with components from traditional kits, which could be egressed outside the capillary by electrochemical pumping. At the tip, femtoliter amounts of the kit components were reacted with the analyte to generate hydrogen peroxide for the electrochemical measurement by the ring electrode. Taking advantage of the nanotip and small volume injection, the nanokit was easily inserted into a single cell to determine the intracellular glucose levels and sphingomyelinase (SMase) activity, which had rarely been achieved. High cellular heterogeneities of these two molecules were observed, showing the significance of the nanokit. Compared with the current methods that use a complicated structural design or surface functionalization for the recognition of the analytes, the nanokit has adapted features of the well-established kits and integrated the kit components and detector in one nanometer-sized capillary, which provides a specific device to characterize the reactivity and concentrations of cellular compounds in single cells.

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Multifunctional carbon-nanotube cellular endoscopes

Cited by 227 publications

References 33 publications

On-Demand Delivery of Single DNA Molecules Using Nanopipets

On-Demand Delivery of Single DNA Molecules Using Nanopipets

Nondestructive nanostraw intracellular sampling for longitudinal cell monitoring

Nanokit for single-cell electrochemical analyses

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