Issue 22, 2014

Enhanced live cell imaging via photonic crystal enhanced fluorescence microscopy

Abstract

We demonstrate photonic crystal enhanced fluorescence (PCEF) microscopy as a surface-specific fluorescence imaging technique to study the adhesion of live cells by visualizing variations in cell–substrate gap distance. This approach utilizes a photonic crystal surface incorporated into a standard microscope slide as the substrate for cell adhesion, and a microscope integrated with a custom illumination source as the detection instrument. When illuminated with a monochromatic light source, angle-specific optical resonances supported by the photonic crystal enable efficient excitation of surface-confined and amplified electromagnetic fields when excited at an on-resonance condition, while no field enhancement occurs when the same photonic crystal is illuminated in an off-resonance state. By mapping the fluorescence enhancement factor for fluorophore-tagged cellular components between on- and off-resonance states and comparing the results to numerical calculations, the vertical distance of labelled cellular components from the photonic crystal substrate can be estimated, providing critical and quantitative information regarding the spatial distribution of the specific components of cells attaching to a surface. As an initial demonstration of the concept, 3T3 fibroblast cells were grown on fibronectin-coated photonic crystals with fluorophore-labelled plasma membrane or nucleus. We demonstrate that PCEF microscopy is capable of providing information about the spatial distribution of cell–surface interactions at the single-cell level that is not available from other existing forms of microscopy, and that the approach is amenable to large fields of view, without the need for coupling prisms, coupling fluids, or special microscope objectives.

Graphical abstract: Enhanced live cell imaging via photonic crystal enhanced fluorescence microscopy

Supplementary files

Article information

Article type
Paper
Submitted
17 Aug 2014
Accepted
18 Sep 2014
First published
18 Sep 2014

Analyst, 2014,139, 5954-5963

Author version available

Enhanced live cell imaging via photonic crystal enhanced fluorescence microscopy

W. Chen, K. D. Long, H. Yu, Y. Tan, J. S. Choi, B. A. Harley and B. T. Cunningham, Analyst, 2014, 139, 5954 DOI: 10.1039/C4AN01508H

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