Cell placement and guidance on substrates for neurochip interfaces

Charrier, Anne; Martínez, Dolores Fernández; Monette, Robert; Comas, Tanya; Movileanu, Raluca; Py, Christophe; Denhoff, M. W.; Krantis, Anthony; Mealing, Geoff

doi:10.1002/bit.22539

Cited by 17 publications

(21 citation statements)

References 20 publications

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“…Since cells can be trapped and detected, we can now proceed to the next step and culture them directly on the PClCA, and thus investigate the possibility of planar patch clamping on cultured cells and, eventually, on neurons. This will require surface functionalization and patterning to ensure adhesion and eventually to guide neuron network establishment (Charrier et al 2010;Bani-Yaghoub et al 2005). The described PClCA also combines local and diffused superfusion of small volumes, hence adhered cells can then be exposed to alternating sequences of buffers and chemicals, drugs and ion channel blockers to study their respective effects on the cultured cells.…”

Section: Discussionmentioning

confidence: 99%

Characterization of a patch-clamp microchannel array towards neuronal networks analysis

Alberti

Snakenborg

Lopacinska

et al. 2010

Microfluid Nanofluid

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The attempt to combine the planar patch clamping idea with the microelectrode array (MEA) concept has led to the fabrication of a patch clamp microchannel array (PClCA). Such a system is thought to be a powerful framework for neuroscience research and drug screening, as a novel tool for simultaneous patch clamping of cultured cells or neurons in the same network. A disposable silicon/silicon dioxide (Si/SiO 2 ) chip with a microhole array was integrated in a microfluidic system for cell handling, perfusion and electrical recording. Fluidic characterization showed that our PClCA can work as a precise local perfusion system for chemicals or drugs. Electrical characterization for microholes of 2 lm and 3 lm revealed an access resistance of 8.09 ± 0.84 MX and 3.18 ± 0.63 MX, respectively. The capacitance was 98.6 ± 13.2 pF. The values are close to what can be expected from theory, but the capacitance is still too high for high resolution recording. The system was tested on HeLa cells: successful cell trapping with a sealing of 40 MX was recorded. Modification of the Si/SiO 2 chip is needed in order to achieve a better sealing and long-term cell culturing in the PClCA remains to be tested.

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Section: Discussionmentioning

confidence: 99%

Characterization of a patch-clamp microchannel array towards neuronal networks analysis

Alberti

Snakenborg

Lopacinska

et al. 2010

Microfluid Nanofluid

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“…Snail neurons, as other large cells, are amenable to manual positioning on top of the probe. For smaller cells requiring longer culture times, we have obviated the need for any manipulation and keep a high probability of obtaining a seal by patterning adhesion polypeptides on top of the probes to place cells on top of the probes, and demonstrated placement of cells on the probe 17,18 .…”

Section: Discussionmentioning

confidence: 99%

Culturing and Electrophysiology of Cells on NRCC Patch-clamp Chips

Martina

Monette

et al. 2012

JoVE

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Due to its exquisite sensitivity and the ability to monitor and control individual cells at the level of ion channels, patch-clamping is the gold standard of electrophysiology applied to disease models and pharmaceutical screens alike 1 . The method traditionally involves gently contacting a cell with a glass pipette filled by a physiological solution in order to isolate a patch of the membrane under its apex 2 . An electrode inserted in the pipette captures ion-channel activity within the membrane patch or, when ruptured, for the whole cell. In the last decade, patch-clamp chips have been proposed as an alternative 3,4 : a suspended film separates the physiological medium from the culture medium, and an aperture microfabricated in the film replaces the apex of the pipette. Patch-clamp chips have been integrated in automated systems and commercialized for high-throughput screening 5 . To increase throughput, they include the fluidic delivery of cells from suspension, their positioning on the aperture by suction, and automated routines to detect cell-to-probe seals and enter into whole cell mode. We have reported on the fabrication of a silicon patch-clamp chip with optimized impedance and orifice shape that permits the high-quality recording of action potentials in cultured snail neurons 6 ; recently, we have also reported progress towards interrogating mammalian neurons 7 . Our patch-clamp chips are fabricated at the Canadian Photonics Fabrication Centre 8 , a commercial foundry, and are available in large series. We are eager to engage in collaborations with electrophysiologists to validate the use of the NRCC technology in different models. The chips are used according to the general scheme represented in Figure 1: the silicon chip is at the bottom of a Plexiglas culture vial and the back of the aperture is connected to a subterranean channel fitted with tubes at either end of the package. Cells are cultured in the vial and the cell on top of the probe is monitored by a measuring electrode inserted in the channel.The two outside fluidic ports facilitate solution exchange with minimal disturbance to the cell; this is an advantage compared to glass pipettes for intracellular perfusion. We detail here the protocols to sterilize and prime the chips, load them with medium, plate them with cells, and finally use them for electrophysiological recordings. Video LinkThe video component of this article can be found at https://www.jove.com/video/3288/

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“…In both cases, photolithography and lCP techniques have been implemented to perform poly-L-lysine or poly-D-lysine micropatterns aligned to microelectrodes arrays with a micrometric accuracy. Successful tests with cortical [132] and hippocampal [133] rodent patterned neuronal networks aged from a few days to several weeks in vitro have been performed, showing for example the propagation of an action potential between two electrode sites distant of 200 lm [131].…”

Section: Instrumentation Of Neuronal Networkmentioning

confidence: 99%