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Direct Imaging of ER Calcium with Targeted-Esterase Induced Dye Loading (TED)
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Biologia
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JoVE Journal Biologia
Direct Imaging of ER Calcium with Targeted-Esterase Induced Dye Loading (TED)

Direct Imaging of ER Calcium with Targeted-Esterase Induced Dye Loading (TED)

DOI:
10.3791/50317-v

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09:32 min

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May 07, 2013

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, , , , ,
1Institute for Clinical Neurobiology,University of Wuerzburg, 2Department of Synapses – Circuits – Plasticity,Max Planck Institute of Neurobiology, Martinsried, 3Walter Brendel Centre of Experimental Medicine,Ludwig-Maximilians University of Munich

Capítulos

  • 00:05Título
  • 02:07Preparation of Live Cell Imaging
  • 02:50Dye Loading of Cells
  • 04:06ER-calcium Live Cell Imaging with an Inverted Laser Scanning Confocal Microscope
  • 05:57Image Processing and Data Analysis
  • 06:38Representative Endoplasmic Reticulum Calcium Imaging Results
  • 08:18Conclusion

Summary

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Targeted-esterase induced dye loading (TED) supports the analysis of intracellular calcium store dynamics by fluorescence imaging. The method bases on targeting of a recombinant Carboxylesterase to the endoplasmic reticulum (ER), where it improves the local unmasking of synthetic low-affinity Ca2+ indicator dyes in the ER lumen.

Tags

Calcium DynamicsER CalciumTEDTargeted-esterase Induced Dye LoadingCalcium IndicatorsFluo5NMag-Fluo4Mag-Fura2CarboxylesteraseLentivirusLive Cell ImagingER StructureCalcium ReleaseCalcium Refilling

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