Multi-dimensional fluorescence microscope

Southwest Sciences, Inc. proposes development of an innovative five-dimensional fluorescence imaging system that combines spectrally resolved, scanning confocal fluorescence microscopy with fluorescence lifetime detection. Our proposed multi-dimensional fluorescence microscope is unique from previous work because it provides simultaneous measurement of lifetime decay profiles at multiple wavelengths. The combination of spectral and lifetime resolution will provide superior contrast between multiple fluorophores and allow the design of more sophisticated fluorescence experiments for structural and functional imaging of cellular and sub-cellular systems. Complete fluorescence decay profiles will be obtained with one nanosecond resolution at each of 16 wavelengths. The proposed microscope and detection instrumentation are a flexible system that is generally applicable to a wide range of fluorescence microscopy applications. The instrument can be designed to accommodate different excitation and emission wavelengths and is compatible with either single or two photon excitation. A modular design that can accept different excitation methods, optics and detectors can be developed. Diode laser excitation and radio frequency (rf) detection electronics will result in a compact, low power, lightweight instrument.