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Time-Lapse Imaging of Neuronal Arborization using Sparse Adeno-Associated Virus Labeling of Genetically Targeted Retinal Cell Populations
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JoVE 신문 신경과학
Time-Lapse Imaging of Neuronal Arborization using Sparse Adeno-Associated Virus Labeling of Genetically Targeted Retinal Cell Populations
DOI:

13:13 min

March 19, 2021

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Chapters

  • 00:04Introduction
  • 00:58Adeno-Associated Virus (AAV) Preparation and Intravitreal Injection of Neonatal Mice
  • 03:13Retinal Dissections for Live-Imaging Experiment
  • 05:04Retinal Flat-Mount Preparation
  • 06:35Time-Lapse Confocal Imaging of Live Whole-Mount Retina Preparations
  • 08:33Image Deconvolution and Post-Processing in ImageJ
  • 11:16Results: Representative Results After Neonatal Intraocular Injection, Time-Lapse Imaging, and Deconvolution
  • 12:13Conclusion

Summary

자동 번역

Here, we present a method for investigating neurite morphogenesis in postnatal mouse retinal explants by time-lapse confocal microscopy. We describe an approach for sparse labeling and acquisition of retinal cell types and their fine processes using recombinant adeno-associated virus vectors that express membrane-targeted fluorescent proteins in a Cre-dependent manner.

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