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Dissection of Local Ca2+ Signals in Cultured Cells by Membrane-targeted Ca2+ Indicators
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Dissection of Local Ca2+ Signals in Cultured Cells by Membrane-targeted Ca2+ Indicators

Dissection of Local Ca2+ Signals in Cultured Cells by Membrane-targeted Ca2+ Indicators

DOI:
10.3791/59246-v

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11:33 min

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March 22, 2019

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, , ,
1Japan Science and Technology Agency,PRESTO, 2Laboratory for Developmental Neurobiology,RIKEN Center for Brain Science, 3École Normale Supèrieure, Institut de Biologie de l’ENS (IBENS), Institut national de la santè et de la recherche mèdicale (INSERM), Centre national de la recherche scientifique (CNRS), École Normale Supèrieure, PSL Research University

Chapters

  • 00:04Title
  • 01:10Preparation of Poly(ethyleneimine) Coated Coverslips
  • 02:42Preparation of Rat Cortical Neuron-astrocyte Mixed Culture and Frozen Cells, and the Revival of Frozen Cultures
  • 05:45Transfection and Adeno-associated Virus Infection of Hippocampal or Cortical Neurons
  • 07:29Recording Spontaneous Activities of Astrocytes Expressing Lck-RCaMP2 and OER-GCaMP6f
  • 08:37Results: Dissection of Local Ca2+ Signals in Cultured Cells
  • 10:38Conclusion

Summary

Automatic Translation

Automatic Translation

Here we present a protocol for Ca2+ imaging in neurons and glial cells, which enables the dissection of Ca2+ signals at subcellular resolution. This process is applicable to all cell types that allow the expression of genetically encoded Ca2+ indicators.

Tags

Keywords: Ca2+ SignalsCalcium ImagingGenetically Encoded Calcium IndicatorsSubcellular ResolutionCalcium InfluxCalcium ReleaseCell CulturePEI CoatingCell DissociationCortexTrypsinDNasePlating Medium

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