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Time-Lapse Video Microscopy for Assessment of EYFP-Parkin Aggregation as a Marker for Cellular Mitophagy
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Biologia
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JoVE Journal Biologia
Time-Lapse Video Microscopy for Assessment of EYFP-Parkin Aggregation as a Marker for Cellular Mitophagy

Time-Lapse Video Microscopy for Assessment of EYFP-Parkin Aggregation as a Marker for Cellular Mitophagy

DOI:
10.3791/53657-v

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09:29 min

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May 04, 2016

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, , ,
1Department of Cancer Biology,Thomas Jefferson University, SKCC, 2Department of Medical Oncology,Thomas Jefferson University, SKCC, 3Kazan Federal University

Capítulos

  • 00:05Título
  • 00:49Electroporation of Fibroblasts with EYFP-Parkin and pDsRed2-Mito Expression Vectors
  • 03:44Time-lapse Video Microscopy
  • 07:20Results: EYFP-Parkin Aggregates in Mitochondrial Membranes During Mitophagy
  • 08:55Conclusion

Summary

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Herein, we describe in detail a time-lapse video microscopy approach to measuring the temporal recruitment of EYFP-Parkin during the selective removal of damaged mitochondria. This dynamic process of EYFP-Parkin-dependent removal of damaged mitochondria can be used as an indicator of cellular health under different experimental conditions.

Tags

Time-lapse Video MicroscopyEYFP-Parkin AggregationMitophagyMolecular MetabolismMitochondrial Life CyclePrimary Cell CulturesEmbryology StudiesImmortalized Mouse Embryonic FibroblastsDMEM MediumTrypsinizationElectroporationEYFP Parkin Expression PlasmidDsRed2 Mito Expression Plasmid

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