Intranuclear immunostaining-based FACS protocol from embryonic cortical tissue

M Sadman Sakib; Godwin Sokpor; Huu Phuc Nguyen; Andre Fischer; Tran Tuoc

doi:10.1016/j.xpro.2021.100318

Intranuclear immunostaining-based FACS protocol from embryonic cortical tissue

STAR Protoc. 2021 Feb 2;2(1):100318. doi: 10.1016/j.xpro.2021.100318. eCollection 2021 Mar 19.

Authors

M Sadman Sakib¹, Godwin Sokpor², Huu Phuc Nguyen², Andre Fischer^{1

3}, Tran Tuoc²

Affiliations

¹ German Center for Neurodegenerative Diseases, 37077 Goettingen, Germany.
² Ruhr University of Bochum, 44791 Bochum, Germany.
³ Cluster of Excellence "Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells (MBExC)," 37077 Goettingen, Germany.

Abstract

Cell sorting can be used to purify cell populations for cell type-specific molecular probing. Fluorescence-activated cell sorting (FACS) coupled with high-throughput sequencing affords molecular signature identification for specific cell types. FACS has many challenges that limit comprehensive cell purification from the brain, leading to incomplete molecular characterization. Here, we present the intranuclear immunostaining-based FACS protocol with several modified steps, which allows optimized nuclei/cell sorting from mouse or human embryonic cortical tissue for distinct downstream molecular investigation of basal intermediate progenitors.

Keywords: Flow Cytometry/Mass Cytometry; Genomics; Neuroscience.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Separation / methods*
Flow Cytometry / methods*
High-Throughput Nucleotide Sequencing
Humans
Immunoassay / methods*
Mice