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Proceedings Paper

Mass measurements of focal adhesions in single cells using high resolution surface plasmon resonance microscopy
Author(s): Alexander W. Peterson; Michael Halter; Alessandro Tona; Anne L. Plant; John T. Elliott
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Paper Abstract

Surface plasmon resonance microscopy (SPRM) is a powerful label-free imaging technique with spatial resolution approaching the optical diffraction limit. The high sensitivity of SPRM to small changes in index of refraction at an interface allows imaging of dynamic protein structures within a cell. Visualization of subcellular features, such as focal adhesions (FAs), can be performed on live cells using a high numerical aperture objective lens with a digital light projector to precisely position the incident angle of the excitation light. Within the cell-substrate region of the SPRM image, punctate regions of high contrast are putatively identified as the cellular FAs. Optical parameter analysis is achieved by application of the Fresnel model to the SPRM data and resulting refractive index measurements are used to calculate protein mass. FAs are known to be regions of high protein density that reside at the cell-substratum interface. Comparing SPRM with fluorescence images of antibody stained for vinculin, a component in FAs, reveals similar measurements of FA size. In addition, a positive correlation between FA size and protein density is revealed by SPRM. Comparing SPRM images for two cell types reveals a distinct difference in the protein density and mass of their respective FAs. Application of SPRM to quantify mass can greatly aid monitoring basic processes that control FA mass and growth and contribute to accurate models that describe cell-extracellular interactions.

Paper Details

Date Published: 23 February 2018
PDF: 7 pages
Proc. SPIE 10509, Plasmonics in Biology and Medicine XV, 1050905 (23 February 2018); doi: 10.1117/12.2290776
Show Author Affiliations
Alexander W. Peterson, National Institute of Standards and Technology (United States)
Michael Halter, National Institute of Standards and Technology (United States)
Alessandro Tona, National Institute of Standards and Technology (United States)
Anne L. Plant, National Institute of Standards and Technology (United States)
John T. Elliott, National Institute of Standards and Technology (United States)

Published in SPIE Proceedings Vol. 10509:
Plasmonics in Biology and Medicine XV
Tuan Vo-Dinh; Joseph R. Lakowicz, Editor(s)

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