Imaging multi-spectral fluorescent probes in living cells at video rates

A fast digital imaging fluorescence microscope capable of obtaining multispectral images at video rates (30/s) has been developed. It integrates commercially available equipment with custom-made circuitry and software to record freeze-frame images of 100-ms duration every 1/30 s. The components involved and the sources of noise have been fully characterized to use their models in offline processing. From the raw images, spatial changes in concentration or distribution can be studied as a function of time. The system will be tested with calcium images obtained by loading single isolated smooth muscle cells with Fura-2 fluorescent dye. The system is capable of generating a movie in which the calcium distribution can be seen changing with time.<
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