Our Application Protocol demonstrates the utility of and consistency in recording action potentials from iCell® Cardiomyocytes using a manual perforated patch clamp method. The protocol provides instructions for culturing the cardiomyocytes, preparing the perforated patch clamp, preparing saline solutions, identifying suitable cardiomyocytes for patch clamp, filling the recording pipette, forming the gigaohm (GΩ) seal, recording action potentials, managing voltage protocol settings, and washing glassware.

To use this protocol with iCell Cardiomyocytes2:

  • Adjust your workflow to accommodate the shorter recovery in culture, enabling you to patch the cells as early as day 4 post thaw..
  • Pay attention to gramicidin, preparing it fresh for best results and at a lower final pipette concentration (1 - 50 μg/ml) than described in the protocol to compare seal stability and time to perforate.
  • Use a lower cell density for patch clamp applications. We recommend a seeding density of 12,000 cells/cm2 (22,800 cells/well in a 24-well plate) to improve the ability to have isolated cells.

For more information about iCell Cardiomyocytes and patch clamp applications, see solutions  00000065A, 00000065B, 00000065C, 00000065D, 00000065E, 00000065F, 00000065G, 00000065H, and 00000065I.

References:

  1. Application Protocol: Measuring Cardiac Electrical Activity: Manual Perforated Patch Clamp