Visualizing Single-Cell Secretion Dynamics with Single-Protein Sensitivity
| Autor: | Vahid Sandoghdar, Michael Aigner, Katharina König, Matthew P. McDonald, André Gemeinhardt, Simon Völkl, Marek Piliarik, Stefanie Schaffer, Andreas Mackensen |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Epstein-Barr Virus Infections Herpesvirus 4 Human Lysis Light Cell Bioengineering Virus Cell Line 03 medical and health sciences 0302 clinical medicine Mediator medicine Extracellular Humans Scattering Radiation Microscopy Interference General Materials Science Secretion B-Lymphocytes biology Chemistry Mechanical Engineering Optical Imaging Dynamics (mechanics) General Chemistry Condensed Matter Physics Cell biology 030104 developmental biology medicine.anatomical_structure Immunoglobulin G 030220 oncology & carcinogenesis biology.protein Single-Cell Analysis Antibody |
| Zdroj: | Nano Letters. 18:513-519 |
| ISSN: | 1530-6992 1530-6984 |
| DOI: | 10.1021/acs.nanolett.7b04494 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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