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Super-resolution chemical imaging via Raman spectroscopy provides a significant ability to simultaneously or pseudosimultaneously monitor numerous label-free analytes while elucidating their spatial distribution on the surface of the sample. However, spontaneous Raman is an inherently weak phenomenon making trace detection and thus superresolution imaging extremely difficult, if not impossible. To circumvent this and allow for trace detection of the few chemical species present in any sub-diffraction limited resolution element of an image, we have developed a surface enhanced Raman scattering (SERS) coherent fiber-optic imaging bundle probe consisting of 30,000 individual fiber elements. When the probes are tapered, etched and coated with metal, they provide circular Raman chemical images of a sample with a field of view of approximately 20μm (i.e. diameter) via the array of 30,000 individual 50 nm fiber elements. An acousto-optic tunable filter is used to rapidly scan or select discrete frequencies for multi- or hyperspectral analysis. Although the 50nm fiber element dimensions of this probe inherently provide spatial resolutions of approximately 100nm, further increases in the spatial resolution can be achieved by using a rapid dithering process. Using this process, additional images are obtained one-half fiber diameter translations in the x- and y- planes. A piezostage drives the movement, providing the accurate and reproducible shifts required for dithering. Optimal probability algorithms are then used to deconvolute the related images producing a final image with a three-fold increase in spatial resolution. This paper describes super-resolution chemical imaging using these probes and the dithering method as well as its potential applications in label-free imaging of lipid rafts and other applications within biology and forensics.
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