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Updated: Mar 8, 2026
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Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
Published on: June 30, 2018
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High-throughput precision measurement of subcellular localization in single cells
Tyler J Burns1, Andreas P Frei2, Pier F Gherardini2
1Department of Cancer Biology, Stanford University School of Medicine, Stanford, California.
Summary
Subcellular Localization Assay (SLA) quantifies protein location in single cells using flow cytometry. This method reveals protein movement within organelles during cell cycle and DNA damage.
Area of Science:
- Cell Biology
- Biotechnology
- Molecular Imaging
Background:
- Flow cytometry typically measures cell surface or internal markers.
- Quantifying protein localization within subcellular compartments at scale is challenging.
- Existing methods lack the throughput for analyzing complex cellular populations.
Purpose of the Study:
- To develop a high-throughput method for quantifying subcellular protein localization.
- To expand flow cytometry capabilities to include spatial and visual information within single cells.
- To enable detailed analysis of protein dynamics in complex cell populations.
Main Methods:
- Developed Subcellular Localization Assay (SLA), an adaptation of Proximity Ligation Assay.
- Integrated SLA with flow cytometry for single-cell analysis.
- Applied SLA to detect nuclear import and intranuclear re-localization of proteins.
Main Results:
- SLA enables quantification of protein localization in thousands of cells per second.
- Successfully detected transcription factor nuclear import across cell subsets.
- Measured intranuclear protein re-localization during the cell cycle and DNA damage response.
Conclusions:
- SLA provides a novel dimension for inquiry and analysis in cell biology.
- This method enhances the capabilities of flow cytometry for studying protein localization.
- SLA facilitates high-throughput investigation of subcellular protein dynamics in complex samples.
