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Neuroscience JoVE (Open Access) Citable · DOI

Culturing and Electrophysiology of Cells on NRCC Patch-clamp Chips

DOI: 10.3791/3288-v
What you'll learn
  • ✓ Sterilize and prime planar patch-clamp chips for electrophysiology
  • ✓ Prepare cells and plate neurons onto microelectrode arrays
  • ✓ Perform whole-cell patch-clamp recordings on cultured neurons
  • ✓ Interpret electrophysiological data from chip-based recordings
Protocol

We show how planar patch-clamp chips fabricated at the National Research Council of Canada are sterilized, primed, loaded with medium, plated with cells, and used for electrophysiological recordings.

Difficulty
advanced
Total time
~3–5 days (chip sterilization/priming ~4 hrs; cell culture plating ~1 day; recordings variable by experiment)
Model organism
Snail neurons (Helix pomatia or similar)
Biosafety
BSL-1

Steps

1
Sterilize, prime, and test patch-clamp chips

Sterilize NRCC planar patch-clamp chips using appropriate protocols, prime with appropriate solutions, and perform electrical testing to verify chip functionality before cell plating.

▶ 02:49
2
Prepare chips in cell biology laboratory

Handle sterilized chips in a controlled cell culture environment, load medium, and prepare the chip surface for cell adhesion and plating.

▶ 04:14
3
Plate snail neurons onto microelectrode arrays

Isolate and suspend snail neurons, then plate them onto the prepared patch-clamp chips at appropriate density for single-cell recordings.

▶ 07:07
4
Perform whole-cell electrophysiological recordings

Position the patch-clamp chip on the recording apparatus and execute whole-cell voltage or current-clamp recordings to measure neuronal electrical activity.

▶ 08:45
5
Analyze representative electrophysiology results

Review and interpret electrophysiological recordings obtained from neurons on NRCC chips, validating data quality and signal characteristics.

▶ 09:09
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