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Isolation of Adult Human Astrocyte Populations from Fresh-Frozen Cortex Using Fluorescence-Activated Nuclei Sorting
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JoVE Journal Neuroscience
Isolation of Adult Human Astrocyte Populations from Fresh-Frozen Cortex Using Fluorescence-Activated Nuclei Sorting

Isolation of Adult Human Astrocyte Populations from Fresh-Frozen Cortex Using Fluorescence-Activated Nuclei Sorting

DOI:
10.3791/62405-v

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08:18 min

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April 16, 2021

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, , , ,
1Department of Pathology,Icahn School of Medicine at Mount Sinai, 2Department of Neuroscience and Friedman Brain Institute,Icahn School of Medicine at Mount Sinai, 3Institutes of Brian Science,Fudan University, 4Department of Psychiatry,Icahn School of Medicine at Mount Sinai

Chapters

  • 00:04Introduction
  • 00:42Frozen Tissue Dissociation and Ultracentrifugation
  • 02:13Fluorescence-Activated Nuclei Sorting (FANS)
  • 04:32Downstream Molecular Analyses
  • 05:58Results: Analysis of Human Astrocyte Populations after FANS
  • 07:40Conclusion

Summary

Automatic Translation

Automatic Translation

We have developed a method that enriches for and isolates human astrocyte populations from fresh-frozen tissue for use in downstream molecular analyses.

Tags

Keywords: Adult Human AstrocyteFluorescence-activated Nuclei SortingFresh-frozen CortexCell IsolationCell Type EnrichmentNeuronOligodendrocyte Progenitor CellSucrose BufferUltracentrifugationNuclei ResuspensionTrypan BlueFluorescence-activated Cell SortingNeuNPAX6DAPIForward ScatterSide ScatterSinglet Nuclei

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