Work

For All the Primate FANS: Optimized Isolation of Nuclei from Frozen Postmortem Primate Brain for Fluorescence-Assisted Nuclei Sorting (FANS)

Year: 2025

Type: article

Abstract: ) cerebral cortex tissue. Isolated nuclei can be input directly into single cell epigenomics protocols like ATAC-seq or can be immunostained for enrichment of neuronal nuclei via fluorescent-activated nuclei sorting (FANS) followed by bulk epigenetic methods like methylome sequencing. We adapted and optimized this protocol based on existing human brain tissue protocols. Our protocol specifically addresses challenges presented by postmortem frozen NHP brain tissue, including high levels of myelin debris and reduced RNA integrity. We include key steps and troubleshooting guidance to improve nuclei quality and sorting outcomes, and we also discuss limitations and considerations for researchers interested in using these methods.

Source: The Yale Journal of Biology and Medicine

DOI: 10.59249/lluj2834

Authors: Elaine E. Guevara, Begün Erbaba, Gregory M. Cresswell, Melissa K. Edler, Chet C. Sherwood

Institutions: Duke University, George Washington University, The University of Texas MD Anderson Cancer Center, Kent State University

Language: English

Topic: Single-cell and spatial transcriptomics

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: diamond

Funders: National Institute on Aging, National Human Genome Research Institute

Awards: G2971354783, G4387471451, Alzheimer's Disease Pathology in a Primate Model