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Imaging of Protein Secretion from a Single Cell Using Plasmonic Substrates

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Abstract

Detecting, imaging, and monitoring cell function on a single-cell basis is very important in the field of immunology research where many molecules are secreted from cells in response to external stimuli including immunization. Here, we introduce substrates with plasmonic nanoparticles and fluorescence microscopy as promising imaging methods for studies on molecular processes controlling cell behavior, particularly secretion of cytokines. We developed a unique composition of silver and silica layers of plasmonic nanostructures which resulted in fluorescence enhancement of more than 200-fold for the ensemble of molecules in the immunoassay. For the proof of concept demonstration, we used primary mouse macrophages and imaged tumor necrosis alpha secretion after stimulation of the cells with lipopolysaccharide. We demonstrate that metal-enhanced fluorescence assay provides imaging capability detection of cytokine secretion from a single cell without extensive biochemical procedures as required with standard methods. In addition, it is demonstrated that cell viability can be controlled during secretion.

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Acknowledgments

This research was supported by NIH grants R21CA147975, HG005090, HG002655, and AI0082299.

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Authors and Affiliations

  1. Center for Fluorescence Spectroscopy, Department of Biochemistry and Molecular Biology, School of Medicine, University of Maryland, 725 W. Lombard St, Baltimore, MD, 21201, USA

    Henryk Szmacinski & Joseph R. Lakowicz

  2. Department of Microbiology and Immunology, School of Medicine, University of Maryland, 685 W. Redwood St, Baltimore, MD, 21201, USA

    Vladimir Toshchakov & Wenji Piao

Authors
  1. Henryk Szmacinski
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  2. Vladimir Toshchakov
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  3. Wenji Piao
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  4. Joseph R. Lakowicz
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Correspondence to Henryk Szmacinski.

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Szmacinski, H., Toshchakov, V., Piao, W. et al. Imaging of Protein Secretion from a Single Cell Using Plasmonic Substrates. BioNanoSci. 3, 30–36 (2013). https://doi.org/10.1007/s12668-013-0076-7

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  • DOI: https://doi.org/10.1007/s12668-013-0076-7

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