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Research
Enhanced Fluorescence Microscope
Using a photonic crystal structure with an optical resonance that matches the wavelength of a laser used to excite fluorescent molecules, we can excite adsorbed fluorescent-tagged biomolecules and cells to emit more light than they would on an ordinary surface. At the same time, a photonic crystal surface can also have a resonance at the wavelength of light given off by fluorescent molecules, so that emitted photons can be directed toward a microscope objective. The enhanced excitation and extraction phenomena can be used to detect fluorescent-tagged proteins and DNA with greater sensitivity than currently-used methods. However, commercially available instrumentation currently represents a significant bottleneck in maximizing the performance of photonic-crystal enhanced fluorescence. We are currently developing a fluorescence microscope-based enhanced fluorescence instrument that provides highly efficient light coupling to and extraction from a photonic crystal. CCD-based imaging enables large-area, high-resolution and high-throughput analysis. The prototype instrument also provides label-free imaging using a nearly identical beam path as that used for enhanced fluorescence. This allows complimentary images to be precisely overlaid in order to provide spatially registered images of enhanced fluorescence and surface-bound molecular density. Furthermore, additional imaging techniques available on the microscope, including brightfield and phase contrast, can also be overlaid. We anticipate these new capabilities will significantly reduce current DNA and protein microarray detection limits and will open the door to new techniques for studying cellular adhesion, motility, and membrane-bound protein expression.
Funding for this project is provided by NSF, NIH, and SRU Biosystems. |
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