@article{a6ef1334cd614fcbb158800987e6a692,
title = "Isolation of mouse brain-infiltrating leukocytes for single cell profiling of epitopes and transcriptomes",
abstract = "High dimensional compositional and transcriptional profiling of heterogeneous brain-infiltrating leukocytes can lead to novel biological and therapeutic discoveries. High-quality single-cell leukocyte preparations are a prerequisite for optimal single cell profiling. Here, we describe a protocol for epitope and RNA-preserving dissociation of adult mouse brains and subsequent leukocyte purification and staining, which is adaptable to homeostatic and pathogenic brains. Leukocyte preparation following this protocol permits exquisite single-cell surface protein and RNA profiling in applications including CyTOF and CITE-seq. For complete details on the use and execution of this protocol, please refer to Guldner et al. (2020) and Golomb et al. (2020).",
keywords = "Cell isolation, Flow Cytometry/Mass Cytometry, Immunology, Neuroscience, RNA-seq, Sequencing, Single Cell",
author = "Guldner, {Ian H.} and Golomb, {Samantha M.} and Qingfei Wang and Emilia Wang and Siyuan Zhang",
note = "Funding Information: This work was funded by NIH R01 CA194697-01 (S.Z.), NIH R01 CA222405 - 01A1 (S.Z.), Notre Dame CRND Catalyst Award (S.Z. and I.H.G.), and NIH CTSI core facility pilot grants (S.Z.). We would like to acknowledge and thank the Dee Family Endowment (S.Z.). We are grateful for the use of the following core facilities and their members{\textquoteright} technical insight: Notre Dame Genomics and Bioinformatics Core Facility, Notre Dame Freimann Life Sciences Center, and Indiana University Simon Cancer Center Core Facility. Funding Information: This work was funded by NIH R01 CA194697-01 (S.Z.), NIH R01 CA222405 - 01A1 (S.Z.), Notre Dame CRND Catalyst Award (S.Z. and I.H.G.), and NIH CTSI core facility pilot grants (S.Z.). We would like to acknowledge and thank the Dee Family Endowment (S.Z.). We are grateful for the use of the following core facilities and their members{\textquoteright} technical insight: Notre Dame Genomics and Bioinformatics Core Facility, Notre Dame Freimann Life Sciences Center, and Indiana University Simon Cancer Center Core Facility. I.H.G. Q.W. and S.Z. designed experimental protocols. I.H.G. S.M.G. Q.W. E.W. and S.Z. performed experiments. I.H.G. S.M.G. E.W. and S.Z. analyzed data. I.H.G. S.M.G. E.W. and S.Z. wrote and revised the protocol. S.Z. supervised the study. All authors reviewed the manuscript. The authors declare no competing interests. Publisher Copyright: {\textcopyright} 2021 The Author(s)",
year = "2021",
month = jun,
day = "18",
doi = "10.1016/j.xpro.2021.100537",
language = "English (US)",
volume = "2",
journal = "STAR Protocols",
issn = "2666-1667",
publisher = "Cell Press",
number = "2",
}