A form of microscopy based on the unique optical properties of a photonic crystal (PC) biosensor provides both label-free (LF) and enhanced fluorescence (EF) imaging modalities for the study of cell-surface interactions and nanoparticle-based assays. The imaging system utilizes an angle-tunable illumination source, either LED or laser, to achieve highly efficient coupling of incident light to PC optical resonances. Detection of the wavelength of resonantly reflected light from PC surfaces enable high-sensitivity label-free biosensing, whereas the enhanced electromagnetic (EM) fields associated with the resonances can be used to strongly improve the fluorescence output of fluorescent labels above PC surfaces. Photonic Crystal Enhanced Microscopy (PCEM) was applied to the study of murine dental stem cells by providing label-free, quantitative, submicron resolution, time-resolved images of the evolution of cell attachment and morphology during chemotaxis and drug-induced apoptosis. It enabled the monitoring of cell behavior with spatial resolution sufficient for observing intracellular attachment strength gradients. PCEM was also used to image fluorescence labeled cells, in which alternating on/off-resonance images was used to derive maps of the surface engagement of cell structures.
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Photonic crystal enhanced microscopy for imaging of live cell adhesion