High‐throughput precision measurement of subcellular localization in single cells

• Published in: Cytometry. Part A Vol. 91; no. 2; pp. 180 - 189
• Main Authors: Burns, Tyler J., Frei, Andreas P., Gherardini, Pier F., Bava, Felice A., Batchelder, Jake E., Yoshiyasu, Yuki, Yu, Julie M., Groziak, Amanda R., Kimmey, Samuel C., Gonzalez, Veronica D., Fantl, Wendy J., Nolan, Garry P.
• Format: Journal Article
• Language: English
• Published: United States Wiley Subscription Services, Inc 01.02.2017
• Subjects: Cell cycle; Cell Nucleus - metabolism; Cytometry; Cytoplasm - metabolism; Cytoplasm - ultrastructure; Deoxyribonucleic acid; DNA; DNA damage; DNA Damage - genetics; DNA damage response; Flow cytometry; Flow Cytometry - methods; High-Throughput Screening Assays - methods; Humans; Localization; nuclear localization; Nuclear transport; Organelles; Protein Transport - genetics; Proteins; proximity ligation assay; Single-Cell Analysis - methods; Subcellular Fractions - ultrastructure; subcellular localization; Transcription factors; nuclear localization; proximity ligation assay; DNA damage response; subcellular localization
• ISSN: 1552-4922; 1552-4930
• DOI: 10.1002/cyto.a.23054

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