Nuclei isolation from surgically resected human hippocampus

Fatma Ayhan; Connor Douglas; Bradley C. Lega; Genevieve Konopka

doi:10.1016/j.xpro.2021.100844

Nuclei isolation from surgically resected human hippocampus

Fatma Ayhan, Connor Douglas, Bradley C. Lega, Genevieve Konopka

Research output: Contribution to journal › Article › peer-review

10 Scopus citations

Abstract

Single-nucleus RNA sequencing (snRNA-seq), where nuclear transcriptomes are a proxy to cellular transcriptomes, has been used to profile human brain. snRNA-seq is sensitive to tissue processing, tissue quality, postmortem interval time, and cellular debris. This protocol outlines steps for the isolation of high-quality nuclei from surgically resected human brain tissue followed by a sucrose gradient yielding neuronal and non-neuronal nuclei enabling unbiased analysis of various cell types. For complete details on the use and execution of this protocol, please refer to Ayhan et al. (2021).

Original language	English (US)
Article number	100844
Journal	STAR Protocols
Volume	2
Issue number	4
DOIs	https://doi.org/10.1016/j.xpro.2021.100844
State	Published - Dec 17 2021

Keywords

Cell isolation
Cell separation/fractionation
Genomics
Microbiology
Neuroscience
RNAseq
Single Cell

ASJC Scopus subject areas

General Immunology and Microbiology
General Biochemistry, Genetics and Molecular Biology
General Neuroscience

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10.1016/j.xpro.2021.100844

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title = "Nuclei isolation from surgically resected human hippocampus",

abstract = "Single-nucleus RNA sequencing (snRNA-seq), where nuclear transcriptomes are a proxy to cellular transcriptomes, has been used to profile human brain. snRNA-seq is sensitive to tissue processing, tissue quality, postmortem interval time, and cellular debris. This protocol outlines steps for the isolation of high-quality nuclei from surgically resected human brain tissue followed by a sucrose gradient yielding neuronal and non-neuronal nuclei enabling unbiased analysis of various cell types. For complete details on the use and execution of this protocol, please refer to Ayhan et al. (2021).",

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AB - Single-nucleus RNA sequencing (snRNA-seq), where nuclear transcriptomes are a proxy to cellular transcriptomes, has been used to profile human brain. snRNA-seq is sensitive to tissue processing, tissue quality, postmortem interval time, and cellular debris. This protocol outlines steps for the isolation of high-quality nuclei from surgically resected human brain tissue followed by a sucrose gradient yielding neuronal and non-neuronal nuclei enabling unbiased analysis of various cell types. For complete details on the use and execution of this protocol, please refer to Ayhan et al. (2021).

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KW - Single Cell

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Nuclei isolation from surgically resected human hippocampus

Abstract

Keywords

ASJC Scopus subject areas

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