DIG FREQ DOMAIN HETERODYNING FLUOR LIFETIME ACQUISITION MODULE

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. FULL TITLE: Digital Frequency Domain Heterodyning Fluorescence Lifetime Acquisition Module for Laser Scanning Microscopy System This project is to design and implement a fluorescence lifetime acquisition module for laser scanning confocal microscopes based on Field Programmable Gate Array (FPGA) technology. The basic challenge of measuring fluorescence lifetime is to accurately determine the time between the excitation and the emission on the nanosecond timescale. The FPGA chip allows the implementation of digital frequency-domain heterodyning on the order of 100MHz to measure fluorescence lifetime decays with an improved duty-cycle and ease of use over analog frequency-domain techniques, and significantly lower cost than time-domain techniques. The principle of the digital frequency domain heterodyning method is described. The operation was implemented in a particular FPGA chip and used in a commercial laser scanning microscope. Preliminary results include the measurement of known lifetime samples to illustrate the sensitivity and noise characteristics of this device. Future work will extend to the biological applications to demonstrate the determination of cellular activities with fluorescence lifetime imaging microscopy (FLIM).