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Isolation of Region-specific Microglia from One Adult Mouse Brain Hemisphere for Deep Single-cell RNA Sequencing
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Neurosciences
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Journal JoVE Neurosciences
Isolation of Region-specific Microglia from One Adult Mouse Brain Hemisphere for Deep Single-cell RNA Sequencing

Isolation of Region-specific Microglia from One Adult Mouse Brain Hemisphere for Deep Single-cell RNA Sequencing

DOI:
10.3791/60347-v

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09:49 min

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December 03, 2019

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,
1Department of Neurobiology,Stanford University School of Medicine

Chapitres

  • 00:05Titre
  • 00:43Brain Region Dissection
  • 01:43Mechanical Tissue Dissociation
  • 02:40Myelin Removal
  • 04:25Staining for Fluorescence-activated Cell Sorting
  • 05:18Index FACS Sorting
  • 05:46Single-cell RNA-sequencing Library Preparation
  • 07:17Results: Isolation of Region-Specific Microglia for Deep Single-Cell RNA Sequencing
  • 09:16Conclusion

Summary

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Traduction automatique

We provide a protocol for isolation of microglia from different dissected regions of an adult mouse brain hemisphere, followed by semi-automated library preparation for deep single-cell RNA sequencing of full-length transcriptomes. This method will help to elucidate functional heterogeneity of microglia in health and disease.

Tags

MicrogliaSingle-cell RNA SequencingBrain RegionsCell IsolationCell SeparationDounce HomogenizerMagnetic SeparationRNA ExtractionLibrary PreparationDeep Sequencing

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