Voltage controlled nano-injection system for single-cell surgery

Seger, R; Actis, Paolo; Penfold, Catherine; Maalouf, Michelle; Vilozny, Boaz; Pourmand, Nader

doi:10.1039/c2nr31700a

Cited by 78 publications

(60 citation statements)

References 30 publications

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“…29 To adapt the SICM as a single-cell biopsy platform, the nanopipette was filled with a 10 mM THATPBCl solution in DCE, and fitted with a silver wire coated with AgTBACl (see online methods). When a DCE-filled nanopipette is immersed into an aqueous solution, a liquid-liquid interface is formed at the nanopore lumen due to the hydrophobic nature of DCE.…”

Section: Resultsmentioning

confidence: 99%

“…22 Our group has developed nanopipettes as a label-free sensing platform 23–28 and we recently described an adaptation of SICM allowing the multi-component injection of single cells in culture. 29 Laforge et al . 30 developed an electrochemical attosyringe based on a glass nanopipette to deliver a minute amount of liquid into a living cell via electrowetting.…”

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confidence: 99%

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Compartmental Genomics in Living Cells Revealed by Single-Cell Nanobiopsy

et al. 2013

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The ability to study the molecular biology of living single cells in heterogeneous cell populations is essential for next generation analysis of cellular circuitry and function. Here, we developed a single-cell nanobiopsy platform based on scanning ion conductance microscopy (SICM) for continuous sampling of intracellular content from individual cells. The nanobiopsy platform uses electrowetting within a nanopipette to extract cellular material from living cells with minimal disruption of the cellular milieu. We demonstrate the subcellular resolution of the nanobiopsy platform by isolating small subpopulations of mitochondria from single living cells, and quantify mutant mitochondrial genomes in those single cells with high throughput sequencing technology. These findings may provide the foundation for dynamic subcellular genomic analysis.

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Compartmental Genomics in Living Cells Revealed by Single-Cell Nanobiopsy

et al. 2013

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“…Additionally, due to (di-)electrophoretic forces, the deposition of molecules could be precisely controlled25. The next step was to inject molecules directly into cells, overcoming the cell membrane similar to microinjection, except that the tips are now about ten times smaller in diameter and the delivery is driven by electrophoretic forces, using typical injection voltages of ± 20 V26. Compared to microinjection, the use of electrophoretic forces instead of pressure driven liquid transfer and the smaller pipette diameters should have a significantly reduced impact on the overall volume displacement inside the cell, as well as the disrupted area of the membrane and cytoplasm of the cell, and therefore its viability.…”

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confidence: 99%

Survival rate of eukaryotic cells following electrophoretic nanoinjection

Simonis

Hübner

Wilking

et al. 2017

Sci Rep

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Insertion of foreign molecules such as functionalized fluorescent probes, antibodies, or plasmid DNA to living cells requires overcoming the plasma membrane barrier without harming the cell during the staining process. Many techniques such as electroporation, lipofection or microinjection have been developed to overcome the cellular plasma membrane, but they all result in reduced cell viability. A novel approach is the injection of cells with a nanopipette and using electrophoretic forces for the delivery of molecules. The tip size of these pipettes is approximately ten times smaller than typical microinjection pipettes and rather than pressure pulses as delivery method, moderate DC electric fields are used to drive charged molecules out of the tip. Here, we show that this approach leads to a significantly higher survival rate of nanoinjected cells and that injection with nanopipettes has a significantly lower impact on the proliferation behavior of injected cells. Thus, we propose that injection with nanopipettes using electrophoretic delivery is an excellent alternative when working with valuable and rare living cells, such as primary cells or stem cells.

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“…njcho@ntu.edu.sg a multi-functional tool for single channel patch-clamping, 9,10 surface patterning or drug delivery, [11][12][13][14][15][16] combined scanning electrochemical microscopy, 17,18 and measuring mechanical properties. [19][20][21] An electrolyte-filled nanopipette, which is pulled from a glass capillary, is used as the probe.…”

Section: Introductionmentioning

confidence: 99%

Alternative configuration scheme for signal amplification with scanning ion conductance microscopy

Kim

Cho

2015

Review of Scientific Instruments

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Scanning Ion Conductance Microscopy (SICM) is an emerging nanotechnology tool to investigate the morphology and charge transport properties of nanomaterials, including soft matter. SICM uses an electrolyte filled nanopipette as a scanning probe and detects current changes based on the distance between the nanopipette apex and the target sample in an electrolyte solution. In conventional SICM, the pipette sensor is excited by applying voltage as it raster scans near the surface. There have been attempts to improve upon raster scanning because it can induce collisions between the pipette sidewalls and target sample, especially for soft, dynamic materials (e.g., biological cells). Recently, Novak et al. demonstrated that hopping probe ion conductance microscopy (HPICM) with an adaptive scan method can improve the image quality obtained by SICM for such materials. However, HPICM is inherently slower than conventional raster scanning. In order to optimize both image quality and scanning speed, we report the development of an alternative configuration scheme for SICM signal amplification that is based on applying current to the nanopipette. This scheme overcomes traditional challenges associated with low bandwidth requirements of conventional SICM. Using our alternative scheme, we demonstrate successful imaging of L929 fibroblast cells and discuss the capabilities of this instrument configuration for future applications.

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Voltage controlled nano-injection system for single-cell surgery

Cited by 78 publications

References 30 publications

Compartmental Genomics in Living Cells Revealed by Single-Cell Nanobiopsy

Compartmental Genomics in Living Cells Revealed by Single-Cell Nanobiopsy

Survival rate of eukaryotic cells following electrophoretic nanoinjection

Alternative configuration scheme for signal amplification with scanning ion conductance microscopy

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