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Quantitative Reflection Imaging of Fixed Aplysia californica Pedal Ganglion Neurons on Nanostructured Plasmonic Crystals
journal contribution
posted on 2016-02-18, 13:40 authored by An-Phong Le, Somi Kang, Lucas B. Thompson, Stanislav
S. Rubakhin, Jonathan V. Sweedler, John A. Rogers, Ralph G. NuzzoStudies
of the interactions between cells and surrounding environment
including cell culture surfaces and their responses to distinct chemical
and physical cues are essential to understanding the regulation of
cell growth, migration, and differentiation. In this work, we demonstrate
the capability of a label-free optical imaging techniquesurface
plasmon resonance (SPR)to quantitatively investigate the relative
thickness of complex biomolecular structures using a nanoimprinted
plasmonic crystal and laboratory microscope. Polyelectrolyte films
of different thicknesses deposited by layer-by-layer assembly served
as the model system to calibrate the reflection contrast response
originating from SPRs. The calibrated SPR system allows quantitative
analysis of the thicknesses of the interface formed between the cell
culture substrate and cellular membrane regions of fixed Aplysia californica pedal ganglion neurons. Bandpass
filters were used to isolate spectral regions of reflected light with
distinctive image contrast changes. Combining of the data from images
acquired using different bandpass filters leads to increase image
contrast and sensitivity to topological differences in interface thicknesses.
This SPR-based imaging technique is restricted in measurable thickness
range (∼100–200 nm) due to the limited plasmonic sensing
volume, but we complement this technique with an interferometric analysis
method. Described here simple reflection imaging techniques show promise
as quantitative methods for analyzing surface thicknesses at nanometer
scale over large areas in real-time and in physicochemical diverse
environments.
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membrane regionsganglion neuronsPolyelectrolyte filmsinterferometric analysis methodnanometer scalereflection contrast responsecell culture surfacescell culture substrateSPR systembiomolecular structuresnanoimprinted plasmonic crystalimage contrast changesinterface thicknessesAplysia californica Pedal Ganglion Neuronsbandpass filterslaboratory microscopesurface thicknessestopological differencesAplysia californicamodel systemreflection imaging techniques show promisecell growthincrease image contrastQuantitative Reflection ImagingBandpass filtersNanostructured Plasmonic CrystalsStudies
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