Abstract
This chapter introduces fluorescence imaging of live cells, defining its various features. An inverted microscope generally allows more flexibility and workspace for the culture and manipulation of live cells than an upright microscope. All lenses for fluorescence imaging should be checked upon delivery for the quality of point-spread function, using fluorescent beads as the sample. The system should include both a mercury arc lamp and a 100-W quartz-halogen lamp for epi-illumination, coupled through a selection mirror to the microscope. Contrary to common practice, the most suitable lamp for fluorescence imaging of live cells is often a 100-W quartz-halogen lamp. The mercury arc lamp should consist of a well shielded housing, power supply, and power supply cable to minimize the potentially damaging electromagnetic wave during ignition. Cooled CCD cameras are used in most fluorescent imaging applications. In choosing a camera, important parameters include quantum efficiency, noise level, pixel size and full-well capacity, and scanning frequency.
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