Nanofountain Probe Electroporation (NFP-E) of Single Cells

Kang, Wonmo; Yavari, Fazel; Minary‐Jolandan, Majid; Giraldo-Vela, Juan P.; Safi, A.; McNaughton, Rebecca L.; Parpoil, Victor; Espinosa, Horacio D.

doi:10.1021/nl400423c

Cited by 106 publications

(159 citation statements)

References 61 publications

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“…The NFP is a cantilever probe, with embedded microchannels, which allows the application of a local electric field when the probe and cell membrane are in contact. Unprecedented transfection efficiency and delivery of DNA, RNA, plasmids and small molecules were achieved with dosage control and very high viability [182]. In addition, other groups demonstrated successful single-cell transfection using polydimethylsiloxane (PDMS)-based microfluidic devices, e.g.…”

Section: Microfluidics Systems For Single-cell Studiesmentioning

confidence: 99%

USNCTAM perspectives on mechanics in medicine

Bao

Bazilevs

Chung

et al. 2014

J. R. Soc. Interface.

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Over decades, the theoretical and applied mechanics community has developed sophisticated approaches for analysing the behaviour of complex engineering systems. Most of these approaches have targeted systems in the transportation, materials, defence and energy industries. Applying and further developing engineering approaches for understanding, predicting and modulating the response of complicated biomedical processes not only holds great promise in meeting societal needs, but also poses serious challenges. This report, prepared for the US National Committee on Theoretical and Applied Mechanics, aims to identify the most pressing challenges in biological sciences and medicine that can be tackled within the broad field of mechanics. This echoes and complements a number of national and international initiatives aiming at fostering interdisciplinary biomedical research. This report also comments on cultural/educational challenges. Specifically, this report focuses on three major thrusts in which we believe mechanics has and will continue to have a substantial impact. (i) Rationally engineering injectable nano/microdevices for imaging and therapy of disease. Within this context, we discuss nanoparticle carrier design, vascular transport and adhesion, endocytosis and tumour growth in response to therapy, as well as uncertainty quantification techniques to better connect models and experiments. (ii) Design of biomedical devices, including point-of-care diagnostic systems, model organ and multi-organ microdevices, and pulsatile ventricular assistant devices. (iii) Mechanics of cellular processes, including mechanosensing and mechanotransduction, improved characterization of cellular constitutive behaviour, and microfluidic systems for single-cell studies.

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Section: Microfluidics Systems For Single-cell Studiesmentioning

confidence: 99%

USNCTAM perspectives on mechanics in medicine

Bao

Bazilevs

Chung

et al. 2014

J. R. Soc. Interface.

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“…Originally demonstrated in bulk on a large amount of cells, this method has been adopted for delivering molecules including DNA, fluorescent dyes, genes and drugs to single cells using nanoscale probes. Micropipettes [70], conductive AFM tips [71] and lithographically fabricated nanofountain pen probes [72] have been applied for single-cell electro poration. The variety of materials injected into cells using this method includes DNA plasmids, proteins fluorescent dyes and genes.…”

Section: +mentioning

confidence: 99%

One-Dimensional Nanoprobes for Single-Cell Studies

et al. 2013

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Owing to variation of individual cells within a population, single-cell studies are of great interest to researchers. Recent developments in nanofabrication technology have made this area increasingly attractive as one-dimensional (1D) nanoscale probes can be manufactured with increasing accuracy. Here, we provide an overview and description of the major designs that have been reported to date. For more details of what applications could be realized and how, based on the probe shapes and designs, we summarize the most recently reported performances of 1D single-cell probes with their advantages and limitations. Minimally invasive probes are required for long-term experiments on single cells. Carbon nanotubes with their unique properties and structure are excellent candidates for multitask robotic intracellular probes. Carbon nanotube-tipped cellular endoscopes are less invasive compared with pipettes or cantilever tips. Advances in nanofabrication techniques have made it possible to produce more consistent nanoscale cellular probes that can capture a variety of information from optical, electrical and chemical signals. In addition, these tools can transfer tiny amounts of fluids and molecular materials in a highly localized fashion for the purpose of analyzing or stimulating a variety of responses at the level of individual cells and even cellular organelles. We conclude with a critical analysis of the current state of the field as well as the major obstacles for further probe development of minimally invasive probes and their widespread use in cell biology.

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“…However, bulk electroporation methods, including nucleofection 35 (modified electroporation) and microporation, suffer from significant disadvantages: i) the entire cell population is exposed to very high voltages, which routinely causes cell death rates of up to 50%, and/or ii) cells need to be suspended during the process. To address these disadvantages while still utilizing electroporation, the Espinosa group developed nanofountain probe electroporation (NFP-E) for single-cell transfection of adherent cells with cell selectivity, dosage control, and high transfection efficiency and viability 36, 37 . This method uses a microfluidic cantilever to apply a localized electric field to an adherent cell for transfection.…”

Section: Introductionmentioning

confidence: 99%

Microfluidic device for stem cell differentiation and localized electroporation of postmitotic neurons

et al. 2014

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New techniques to deliver of nucleic acids and other molecules for gene editing and gene expression profiling, which can be performed with minimal perturbation to cell growth or differentiation, are essential for advancing biological research. Studying cells in their natural state, with temporal control, is particularly important for primary cells that are derived by differentiation from stem cells and are adherent, e.g., neurons. Existing high-throughput transfection methods either require cells to be in suspension or are highly toxic and limited to a single transfection per experiment. Here we present a microfluidic device that couples on-chip culture of adherent cells and transfection by localized electroporation. Integrated microchannels allow long-term cell culture on the device and repeated temporal transfection. The microfluidic device was validated by first performing electroporation of HeLa and HT1080 cells, with transfection efficiencies of ~95% for propidium iodide and up to 50% for plasmids. Application to primary cells was demonstrated by on-chip differentiation of neural stem cells and transfection of postmitotic neurons with a green fluorescent protein plasmid.

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Nanofountain Probe Electroporation (NFP-E) of Single Cells

Cited by 106 publications

References 61 publications

USNCTAM perspectives on mechanics in medicine

USNCTAM perspectives on mechanics in medicine

One-Dimensional Nanoprobes for Single-Cell Studies

Microfluidic device for stem cell differentiation and localized electroporation of postmitotic neurons

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