Nuclei isolation, immunostaining, and Fluorescence-activated nuclei sorting (FANS) for Smart-Seq2

Output Details

This protocol describes the steps for extracting nuclei from human postmortem brain samples, immunofluorescence, and nuclei sorting (FANS) for single-nucleus RNA-seq using the Smart-Seq2 method. We have used it to isolate nuclei from human brain samples (such as the cingulate cortex), but it can be adapted for nuclei from different body areas, cell culture materials, and/or different species.
Tags
  • Immunofluorescence
  • Nuclei isolation
  • scRNAseq (Single-cell RNA-seq)

Meet the Authors

  • Ester Kalef-Ezra, PhD

    Key Personnel: Team Voet

    University College London

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    Dominic Horner, BSc

    Key Personnel: Team Voet

    University College London

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    George Morrow

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    Vanda Knitlhoffer

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    Andy Goldson

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    Iain Maculay

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    Yanping Guo

  • Christos Proukakis, PhD

    Co-PI (Core Leadership): Team Voet

    University College London