Compartmental Genomics in Living Cells Revealed by Single-Cell Nanobiopsy

Actis, Paolo; Maalouf, Michelle; Kim, Hyunsung John; Lohith, Akshar; Vilozny, Boaz; Seger, R; Pourmand, Nader

doi:10.1021/nn405097u

Cited by 156 publications

(248 citation statements)

References 38 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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“…22 Additionally, nanopipette probes still hold great promise as intracellular biosensors [23][24][25] and as tools for cell manipulation. [26][27][28] We recently developed a method to fabricate multifunctional nanoprobes starting from double-barrel quartz nanopipettes. Pyrolytic carbon is selectively deposited on one barrel while leaving the other one unchanged.…”

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confidence: 99%

Electrochemical Nanoprobes for Single-Cell Analysis

et al. 2014

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The measurement of key molecules in individual cells with minimal disruption to the biological milieu is the next frontier in single-cell analyses. Nanoscale devices are ideal analytical tools because of their small size and their potential for high spatial and temporal resolution recordings. Here, we report the fabrication of disk-shaped carbon nanoelectrodes whose radius can be precisely tuned within the range 5-200 nm. The functionalization of the nanoelectrode with platinum allowed the monitoring of oxygen consumption outside and inside a brain slice. Furthermore, we show that nanoelectrodes of this type can be used to impale individual cells to perform electrochemical measurements within the cell with minimal disruption to cell function. These nanoelectrodes can be fabricated combined with scanning ion conductance microcopy probes which should allow high resolution electrochemical mapping of species on or in living cells.

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“…As well as being able to deliver material to a surface, the SICM nanopipette also allows singlecell nano-biopsies to be performed, through the extraction of a small amount of material from inside a cell which is then further analysed [83,84]. This material can be, for example, mRNA, the levels of which can be quantified using PCR.…”

Section: (D) Conductance Measurements Local Delivery and Sizingmentioning

confidence: 99%

Multifunctional scanning ion conductance microscopy

2017

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Scanning ion conductance microscopy (SICM) is a nanopipette-based technique that has traditionally been used to image topography or to deliver species to an interface, particularly in a biological setting. This article highlights the recent blossoming of SICM into a technique with a much greater diversity of applications and capability that can be used either standalone, with advanced control (potential-time) functions, or in tandem with other methods. SICM can be used to elucidate functional information about interfaces, such as surface charge density or electrochemical activity (ion fluxes). Using a multi-barrel probe format, SICM-related techniques can be employed to deposit nanoscale three-dimensional structures and further functionality is realized when SICM is combined with scanning electrochemical microscopy (SECM), with simultaneous measurements from a single probe opening up considerable prospects for multifunctional imaging. SICM studies are greatly enhanced by finite-element method modelling for quantitative treatment of issues such as resolution, surface charge and (tip) geometry effects. SICM is particularly applicable to the study of living systems, notably single cells, although applications extend to materials characterization and to new methods of printing and nanofabrication. A more thorough understanding of the electrochemical principles and properties of SICM provides a foundation for significant applications of SICM in electrochemistry and interfacial science.

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Compartmental Genomics in Living Cells Revealed by Single-Cell Nanobiopsy

Cited by 156 publications

References 38 publications

On-Demand Delivery of Single DNA Molecules Using Nanopipets

On-Demand Delivery of Single DNA Molecules Using Nanopipets

Electrochemical Nanoprobes for Single-Cell Analysis

Multifunctional scanning ion conductance microscopy

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