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Fluorescence-Activated Nuclei Negative Sorting of Neurons Combined with Single Nuclei RNA Sequencing to Study the Hippocampal Neurogenic Niche
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Fluorescence-Activated Nuclei Negative Sorting of Neurons Combined with Single Nuclei RNA Sequencing to Study the Hippocampal Neurogenic Niche

Fluorescence-Activated Nuclei Negative Sorting of Neurons Combined with Single Nuclei RNA Sequencing to Study the Hippocampal Neurogenic Niche

DOI:
10.3791/64369-v

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08:16 min

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October 20, 2022

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, , , ,
1The Francis Crick Institute, 2MSD R&D Innovation Centre

Chapters

  • 00:04Introduction
  • 01:08Dissection of the Dentate Gyrus (DG)
  • 01:53Tissue Dissociation, Single Nuclei Isolation, and Anti-NeuN Immunostaining
  • 04:03Fluorescence Activated Nuclei Sorting (FANS) to Exclude Neuronal Populations
  • 05:12Preparation of the Single Nuclei Suspension to Perform Single Nuclei RNA Sequencing
  • 05:59Results: Preparation of a Suspension of Non-Neuronal Single Nuclei Isolated from the Dentate Gyrus (DG)
  • 07:51Conclusion

Summary

Automatic Translation

Automatic Translation

Presented here is a method to sequence single nuclei isolated from the mouse dentate gyrus that excludes most neurons through fluorescence-activated nuclei (FAN)-sorting. This approach generates high-quality expression profiles and facilitates the study of most other cell types represented in the niche, including scarce populations such as neural stem cells.

Tags

Keywords: Fluorescence-Activated Nuclei Negative SortingSingle Nuclei RNA SequencingHippocampal Neurogenic NicheNeural Stem CellsAstrocytesOligodendrocytesAdult Hippocampal NeurogenesisDentate GyrusBrain DissectionDounce HomogenizerNuclei SuspensionCell StrainerCentrifugation

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