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Nano Lett. 2018 Jan 10;18(1):513-519. doi: 10.1021/acs.nanolett.7b04494. Epub 2017 Dec 19.

Visualizing Single-Cell Secretion Dynamics with Single-Protein Sensitivity.

Author information

1
Nano-Optics Division, Max Planck Institute for the Science of Light , Staudtstraße 2, 91058 Erlangen, Germany.
2
Department of Physics , Friedrich Alexander University Erlangen-Nuremberg , Schloßplatz 4, 91054 Erlangen, Germany.
3
Department of Internal Medicine 5, Hematology and Oncology, Friedrich Alexander University Erlangen-Nuremberg , Ulmenweg 18, 91054 Erlangen, Germany.

Abstract

Cellular secretion of proteins into the extracellular environment is an essential mediator of critical biological mechanisms, including cell-to-cell communication, immunological response, targeted delivery, and differentiation. Here, we report a novel methodology that allows for the real-time detection and imaging of single unlabeled proteins that are secreted from individual living cells. This is accomplished via interferometric detection of scattered light (iSCAT) and is demonstrated with Laz388 cells, an Epstein-Barr virus (EBV)-transformed B cell line. We find that single Laz388 cells actively secrete IgG antibodies at a rate of the order of 100 molecules per second. Intriguingly, we also find that other proteins and particles spanning ca. 100 kDa-1 MDa are secreted from the Laz388 cells in tandem with IgG antibody release, likely arising from EBV-related viral proteins. The technique is general and, as we show, can also be applied to studying the lysate of a single cell. Our results establish label-free iSCAT imaging as a powerful tool for studying the real-time exchange between cells and their immediate environment with single-protein sensitivity.

KEYWORDS:

cellular secretion; dynamics; iSCAT; imaging; label-free; single-protein

PMID:
29227108
DOI:
10.1021/acs.nanolett.7b04494
[Indexed for MEDLINE]

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