Acquisition of murine splenic myeloid cells for protein and gene expression profiling by advanced flow cytometry and CITE-seq

Inga Rødahl; James Gotley; Stacey B Andersen; Meihua Yu; Ahmed M Mehdi; Angelika N Christ; Emma E Hamilton-Williams; Ian H Frazer; Samuel W Lukowski; Janin Chandra

doi:10.1016/j.xpro.2021.100842

Acquisition of murine splenic myeloid cells for protein and gene expression profiling by advanced flow cytometry and CITE-seq

STAR Protoc. 2021 Sep 17;2(4):100842. doi: 10.1016/j.xpro.2021.100842. eCollection 2021 Dec 17.

Authors

Affiliations

¹ Center for Infectious Medicine, Department of Medicine, Karolinska Institute, Karolinska University Hospital Huddinge, 141 86 Stockholm, Sweden.
² The University of Queensland Diamantina Institute, The University of Queensland, Woolloongabba, QLD 4102, Australia.
³ The Institute for Molecular Bioscience, The University of Queensland, Woolloongabba, QLD 4067, Australia.

Abstract

Here, we outline detailed protocols to isolate and profile murine splenic dendritic cells (DCs) through advanced flow cytometry of the myeloid compartment and single-cell transcriptomic profiling with integrated cell surface protein expression through CITE-seq. This protocol provides a general transferrable road map for different tissues and species. For complete details on the use and execution of this protocol, please refer to Lukowski et al. (2021).

Keywords: Cell isolation; Flow Cytometry/Mass Cytometry; Health Sciences; High Throughput Screening; Immunology; Molecular Biology; Sequence analysis; Sequencing; Single Cell.

Publication types

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

MeSH terms

Animals
Flow Cytometry / methods
Gene Expression Profiling*
Membrane Proteins
Mice
Microarray Analysis
Myeloid Cells*

Substances

Membrane Proteins