FACS protocol for direct comparison of cell populations isolated from mice

Katarzyna A Ludwik; Zachary M Sandusky; Eric B Wright; Deborah A Lannigan

doi:10.1016/j.xpro.2020.100270

FACS protocol for direct comparison of cell populations isolated from mice

STAR Protoc. 2021 Jan 13;2(1):100270. doi: 10.1016/j.xpro.2020.100270. eCollection 2021 Mar 19.

Authors

Katarzyna A Ludwik¹, Zachary M Sandusky¹, Eric B Wright², Deborah A Lannigan^{1

3

2}

Affiliations

¹ Department of Pathology, Microbiology & Immunology, Vanderbilt University Medical Center, Vanderbilt University, Nashville, TN 37232, USA.
² Department of Biomedical Engineering, Vanderbilt University, Nashville, TN 37232, USA.
³ Department of Cell and Developmental Biology, Vanderbilt University, Nashville, TN 37232, USA.

Abstract

A FACS protocol is described that eliminates isolation and staining artifacts to allow accurate comparison between cell populations isolated from organs obtained from disparate mouse groups. This protocol was validated by characterizing the estrogen receptor positive cells within the mammary gland of transgenic mice with different genotypes at different stages of the estrous cycle. We include protocols necessary to batch stage animals within the cycle to proceed directly to FACS, which provides optimal RNA yields for RNA-seq. For complete details on the use and execution of this protocol, please refer to Ludwik et al. (2020).

Keywords: Cell biology; Cell isolation; Flow cytometry/mass cytometry; Genomics; Model organisms.

Publication types

Comparative Study
Research Support, N.I.H., Extramural

MeSH terms

Animals
Estrous Cycle*
Female
Flow Cytometry*
Mammary Glands, Animal / cytology*
Mammary Glands, Animal / metabolism*
Mice
Mice, Transgenic
RNA-Seq

Abstract

Publication types

MeSH terms

Grants and funding