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Download fileQuantitative Reflection Imaging for the Morphology and Dynamics of Live Aplysia californica Pedal Ganglion Neurons Cultured on Nanostructured Plasmonic Crystals
journal contribution
posted on 2017-02-24, 00:00 authored by Somi Kang, Adina Badea, Stanislav S. Rubakhin, Jonathan V. Sweedler, John A. Rogers, Ralph G. NuzzoWe describe a reflection imaging
system that consists of a plasmonic
crystal, a common laboratory microscope, and band-pass filters for
use in the quantitative imaging and in situ monitoring of live cells
and their substrate interactions. Surface plasmon resonance (SPR)
provides a highly sensitive method to monitor changes in physicochemical
properties occurring at metal–dielectric interfaces. Polyelectrolyte
thin films deposited using the layer-by-layer (LBL) self-assembly
method provide a reference system for calibrating the reflection contrast
changes that occur when the polyelectrolyte film thickness changes
and provide insight into the optical responses that originate from
the multiple plasmonic features supported by this imaging system.
Finite-difference time-domain (FDTD) simulations of the optical responses
measured experimentally from the polyelectrolyte reference system
are used to provide a calibration of the optical system for subsequent
use in quantitative studies investigating live cell dynamics in cultures
supported on a plasmonic crystal substrate. Live Aplysia californica pedal ganglion neurons cultured in artificial seawater were used
as a model system through which to explore the utility of this plasmonic
imaging technique. Here, the morphology of cellular peripheral structures
≲80 nm in thickness were quantitatively analyzed, and the dynamics
of their trypsin-induced surface detachment were visualized. These
results illustrate the capacities of this system for use in investigations
of the dynamics of ultrathin cellular structures within complex bioanalytical
environments.
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Keywords
ganglion neuronsreflection imaging systemSPRLBLreference systemreflection contrast changespolyelectrolyte film thickness changespolyelectrolyte reference systemFinite-difference time-domainLive Aplysia californica Pedal Ganglion Neurons CulturedLive Aplysia californicaimaging systemFDTDlaboratory microscopecell dynamicsplasmonic featuresband-pass filtersphysicochemical propertiesQuantitative Reflection Imagingsubstrate interactionsplasmonic imaging techniqueNanostructured Plasmonic Crystalstrypsin-induced surface detachmentplasmonic crystalplasmonic crystal substratebioanalytical environmentsmodel systemSurface plasmon resonanceself-assembly method