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Related Experiment Video

Updated: Oct 18, 2025

Neuronal Nuclei Isolation from Human Postmortem Brain Tissue
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Neuronal Nuclei Isolation from Human Postmortem Brain Tissue

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Nuclei isolation from surgically resected human hippocampus.

Fatma Ayhan1, Connor Douglas1, Bradley C Lega2

  • 1Department of Neuroscience, UT Southwestern Medical Center, Dallas, TX 75390, USA.

STAR Protocols
|September 29, 2021
PubMed
Summary
This summary is machine-generated.

This study details a protocol for isolating high-quality nuclei from human brain tissue for single-nucleus RNA sequencing (snRNA-seq). This method enables unbiased analysis of diverse neuronal and non-neuronal cell types.

Keywords:
Cell isolationCell separation/fractionationGenomicsMicrobiologyNeuroscienceRNAseqSingle Cell

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Genomics

Background:

  • Single-nucleus RNA sequencing (snRNA-seq) is a powerful tool for profiling the human brain.
  • However, snRNA-seq is sensitive to tissue processing, quality, postmortem interval, and cellular debris, impacting data accuracy.

Purpose of the Study:

  • To present a detailed protocol for isolating high-quality nuclei from human brain tissue.
  • To enable unbiased analysis of various neuronal and non-neuronal cell types using snRNA-seq.

Main Methods:

  • Isolation of high-quality nuclei from surgically resected human brain tissue.
  • Utilizing a sucrose gradient to separate neuronal and non-neuronal nuclei.
  • Application of these nuclei for single-nucleus RNA sequencing.

Main Results:

  • Successful isolation of high-quality nuclei from human brain tissue.
  • Separation of neuronal and non-neuronal nuclei populations.
  • Enabling comprehensive and unbiased cell-type analysis.

Conclusions:

  • The described protocol effectively yields high-quality nuclei for snRNA-seq.
  • This method supports detailed characterization of human brain cell populations.
  • It provides a foundation for advanced neurogenomic studies.