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Biophys J. 2013 Aug 6;105(3):602-8. doi: 10.1016/j.bpj.2013.06.022.

Quantitative imaging of protein secretions from single cells in real time.

Author information

1
Materials Science and Technology Division, Naval Research Laboratory, Washington DC, USA. marc.raphael@nrl.navy.mil

Abstract

Protein secretions from individual cells create spatially and temporally varying concentration profiles in the extracellular environment, which guide a wide range of biological processes such as wound healing and angiogenesis. Fluorescent and colorimetric probes for the detection of single cell secretions have time resolutions that range from hours to days, and as a result, little is known about how individual cells may alter their protein secretion rates on the timescale of minutes or seconds. Here, we present a label-free technique based upon nanoplasmonic imaging, which enabled the measurement of individual cell secretions in real time. When applied to the detection of antibody secretions from single hybridoma cells, the enhanced time resolution revealed two modes of secretion: one in which the cell secreted continuously and another in which antibodies were released in concentrated bursts that coincided with minute-long morphological contractions of the cell. From the continuous secretion measurements we determined the local concentration of antibodies at the sensing array closest to the cell and from the bursts we estimated the diffusion constant of the secreted antibodies through the extracellular media. The design also incorporates transmitted light and fluorescence microscopy capabilities for monitoring cellular morphological changes and intracellular fluorescent labels. We anticipate that this technique can be adapted as a general tool for the quantitative study of paracrine signaling in both adherent and nonadherent cell lines.

PMID:
23931308
PMCID:
PMC3736659
DOI:
10.1016/j.bpj.2013.06.022
[Indexed for MEDLINE]
Free PMC Article

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