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Whole-cell Patch-clamp Recordings for Electrophysiological Determination of Ion Selectivity in Channelrhodopsins
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Neuroscienze
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JoVE Journal Neuroscienze
Whole-cell Patch-clamp Recordings for Electrophysiological Determination of Ion Selectivity in Channelrhodopsins

Whole-cell Patch-clamp Recordings for Electrophysiological Determination of Ion Selectivity in Channelrhodopsins

DOI:
10.3791/55497-v

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08:39 min

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May 22, 2017

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, , ,
1Experimental Biophysics, Institute of Biology,Humboldt-Universität zu Berlin

Capitoli

  • 00:05Titolo
  • 00:50Setup Prior to Recordings
  • 01:35Seeding and Transfection of Cells
  • 02:50Selectivity Measurements
  • 06:48Results: Chloride Selectivity of PsACR1
  • 07:51Conclusion

Summary

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This article describes how the ion selectivity of channelrhodopsin is determined with electrophysiological whole-cell patch-clamp recordings using HEK293 cells. Here, the experimental procedure for investigating chloride selectivity of an anion-selective channelrhodopsin is demonstrated. However, the procedure is transferable to other channelrhodopsins of distinct selectivity.

Tags

Keywords: Whole-cell Patch-clampElectrophysiologyIon SelectivityChannelrhodopsinsHEK CellsTransfectionPhotocurrentBiophysicsIntracellular BufferExtracellular BufferOsmolarityPatch PipetteFire PolishingRecording Chamber

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