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荧光激活神经元核负分选结合单核RNA测序研究海马神经源性生态位
JoVE Journal
Neuroscience
This content is Free Access.
JoVE Journal Neuroscience
Fluorescence-Activated Nuclei Negative Sorting of Neurons Combined with Single Nuclei RNA Sequencing to Study the Hippocampal Neurogenic Niche
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荧光激活神经元核负分选结合单核RNA测序研究海马神经源性生态位

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

这里介绍的是从小鼠齿状回分离的单个细胞核测序的方法，该方法通过荧光激活细胞核（FAN）分选排除大多数神经元。这种方法可生成高质量的表达谱，并促进对生态位中代表的大多数其他细胞类型的研究，包括神经干细胞等稀缺群体。

Tags

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

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