, Volume 3, Issue 1, pp 30–36 | Cite as

Imaging of Protein Secretion from a Single Cell Using Plasmonic Substrates

  • Henryk SzmacinskiEmail author
  • Vladimir Toshchakov
  • Wenji Piao
  • Joseph R. Lakowicz


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.


Plasmonic nanostructures Silver nanoparticles Metal-enhanced fluorescence Microphage cells Cell assays Tumor necrosis factor alpha 



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


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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Henryk Szmacinski
    • 1
    Email author
  • Vladimir Toshchakov
    • 2
  • Wenji Piao
    • 2
  • Joseph R. Lakowicz
    • 1
  1. 1.Center for Fluorescence Spectroscopy, Department of Biochemistry and Molecular Biology, School of MedicineUniversity of MarylandBaltimoreUSA
  2. 2.Department of Microbiology and Immunology, School of MedicineUniversity of MarylandBaltimoreUSA

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