title: High-throughput precision measurement of subcellular localization in single cells.
abstract: To quantify visual and spatial information in single cells with a throughput of thousands of cells per second, we developed Subcellular Localization Assay (SLA). This adaptation of Proximity Ligation Assay expands the capabilities of flow cytometry to include data relating to localization of proteins to and within organelles. We used SLA to detect the nuclear import of transcription factors across cell subsets in complex samples. We further measured intranuclear re-localization of target proteins across the cell cycle and upon DNA damage induction. SLA combines multiple single-cell methods to bring about a new dimension of inquiry and analysis in complex cell populations. © 2017 International Society for Advancement of Cytometry.
authors.affiliation: Department of Cancer Biology, Stanford University School of Medicine, Stanford, California.
authors.forename: Tyler J
authors.initials: TJ
authors.lastname: Burns
authors.affiliation: Stanford University School of Medicine, Baxter Laboratory for Stem Cell Biology, Stanford, California.
authors.forename: Andreas P
authors.initials: AP
authors.lastname: Frei
authors.affiliation: Stanford University School of Medicine, Baxter Laboratory for Stem Cell Biology, Stanford, California.
authors.forename: Pier F
authors.initials: PF
authors.lastname: Gherardini
authors.affiliation: Stanford University School of Medicine, Baxter Laboratory for Stem Cell Biology, Stanford, California.
authors.forename: Felice A
authors.initials: FA
authors.lastname: Bava
authors.affiliation: Immunology and Microbial Pathogenesis, Joan and Sanford I. Weill Medical College of Cornell University, New York, New York.
authors.forename: Jake E
authors.initials: JE
authors.lastname: Batchelder
authors.affiliation: Department of Medicine, University of Texas Health Science Center at San Antonio, San Antonio, Texas.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Yoshiyasu
authors.affiliation: Department of Biological Sciences, University of California Berkeley, Berkeley, California.
authors.forename: Julie M
authors.initials: JM
authors.lastname: Yu
authors.affiliation: Department of Biology, Barnard College, New York, New York.
authors.forename: Amanda R
authors.initials: AR
authors.lastname: Groziak
authors.affiliation: Developmental Biology, Stanford University School of Medicine, Stanford, California.
authors.forename: Samuel C
authors.initials: SC
authors.lastname: Kimmey
authors.affiliation: Stanford University School of Medicine, Baxter Laboratory for Stem Cell Biology, Stanford, California.
authors.forename: Veronica D
authors.initials: VD
authors.lastname: Gonzalez
authors.affiliation: Stanford Comprehensive Cancer Institute and Department of Obstetrics and Gynecology, Stanford University, Stanford, California.
authors.forename: Wendy J
authors.initials: WJ
authors.lastname: Fantl
authors.affiliation: Department of Microbiology and Immunology, Stanford University, Stanford, California.
authors.forename: Garry P
authors.initials: GP
authors.lastname: Nolan
mesh_terms: D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism / Q000648:ultrastructure; D004249:DNA Damage / Q000235:genetics; D005434:Flow Cytometry / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D021381:Protein Transport / Q000235:genetics; D059010:Single-Cell Analysis / Q000379:methods*; D013347:Subcellular Fractions / Q000648:ultrastructure
keywords: DNA damage response; nuclear localization; proximity ligation assay; subcellular localization