Fluorescence-activated nuclei sorting (FANS) on human post-mortem cortex tissue enabling the isolation of distinct neural cell populations for multiple omic profiling v2
- 1University of Exeter , Exeter , UK ;
- 2University of Exeter Medical School
- 3University of Exeter Complex Disease Epige… UK Human Functional G…
Abstract
Increased understanding of the functional complexity of the genome has led to growing recognition about the role of epigenetic/transcriptional variation in health and disease. Current analyses of the human brain, however, are limited by the use of “bulk” tissue, comprising a heterogeneous mix of different neural cell types. As epigenetic processes play a critical role in determining cell type-specific patterns of gene regulation it is important to consider cellular composition in regulatory genomic studies of human post-mortem tissue, and there is a need for methods to purify populations of specific cell-types. Furthermore, the valuable nature of human post-mortem tissue means it is important to use methods that maximize the amount of genomic data generated on each sample. This protocol describes a method that uses fluorescence-activated nuclei sorting (FANS) to isolate and profile nuclei from multiple different human brain cell-types from frozen post-mortem tissue. This protocol can be used to robustly purify populations of neuronal (NeuN+ve), oligodendrocytes (SOX10+ve), microglia (IRF8+ve) and other glial origin nuclei (NeuN-ve/SOX10-ve/IRF8-ve) from adult post-mortem frozen brain, with each tissue sample yielding purified populations of nuclei amenable to simultaneous analysis of i) DNA modifications (via bisulfite sequencing / array), ii) histone modifications (CUT&Tag), iii) open chromatin analysis (ATAC-seq), and iv) gene expression (snRNA-seq).
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BibTeX
@misc{policicchio2026fluorescenceactivated,
title = {{Fluorescence-activated nuclei sorting (FANS) on human post-mortem cortex tissue enabling the isolation of distinct neural cell populations for multiple omic profiling v2}},
author = {Stefania S S Policicchio and Barry A. Chioza and Jonathan P. Davies and Joe Burrage and Jonathan S. Mill and Emma Louise Dempster},
year = {2026},
month = may,
doi = {10.17504/protocols.io.36wgq4965vk5/v3},
url = {https://doi.org/10.17504/protocols.io.36wgq4965vk5/v3},
note = {CC BY},
}
RIS
TY - UNPB
TI - Fluorescence-activated nuclei sorting (FANS) on human post-mortem cortex tissue enabling the isolation of distinct neural cell populations for multiple omic profiling v2
AU - Stefania S S Policicchio
AU - Barry A. Chioza
AU - Jonathan P. Davies
AU - Joe Burrage
AU - Jonathan S. Mill
AU - Emma Louise Dempster
PY - 2026
DA - 2026/05/27
DO - 10.17504/protocols.io.36wgq4965vk5/v3
UR - https://doi.org/10.17504/protocols.io.36wgq4965vk5/v3
AB - Increased understanding of the functional complexity of the genome has led to growing recognition about the role of epigenetic/transcriptional variation in health and disease. Current analyses of the human brain, however, are limited by the use of “bulk” tissue, comprising a heterogeneous mix of different neural cell types. As epigenetic processes play a critical role in determining cell type-specific patterns of gene regulation it is important to consider cellular composition in regulatory genomic studies of human post-mortem tissue, and there is a need for methods to purify populations of specific cell-types. Furthermore, the valuable nature of human post-mortem tissue means it is important to use methods that maximize the amount of genomic data generated on each sample. This protocol describes a method that uses fluorescence-activated nuclei sorting (FANS) to isolate and profile nuclei from multiple different human brain cell-types from frozen post-mortem tissue. This protocol can be used to robustly purify populations of neuronal (NeuN+ve), oligodendrocytes (SOX10+ve), microglia (IRF8+ve) and other glial origin nuclei (NeuN-ve/SOX10-ve/IRF8-ve) from adult post-mortem frozen brain, with each tissue sample yielding purified populations of nuclei amenable to simultaneous analysis of i) DNA modifications (via bisulfite sequencing / array), ii) histone modifications (CUT&Tag), iii) open chromatin analysis (ATAC-seq), and iv) gene expression (snRNA-seq).
ER -