High-throughput precision measurement of subcellular localization in single cells

Cytometry A. 2017 Feb;91(2):180-189. doi: 10.1002/cyto.a.23054. Epub 2017 Jan 17.

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.

Keywords: DNA damage response; nuclear localization; proximity ligation assay; subcellular localization.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.
  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Cell Nucleus / metabolism
  • Cytoplasm / metabolism
  • Cytoplasm / ultrastructure
  • DNA Damage / genetics
  • Flow Cytometry / methods*
  • High-Throughput Screening Assays / methods*
  • Humans
  • Protein Transport / genetics
  • Single-Cell Analysis / methods*
  • Subcellular Fractions / ultrastructure