An Introduction to Electronic Image Acquisition during Light Microscopic Observation of Biological Specimens

Publisher Summary This chapter introduces the reader to the choices and considerations that need to be made when designing an image acquisition system. It focuses on the use of electronic cameras to record images of biological specimens generated with light microscopy techniques. The ideal image acquisition system would be sensitive enough to acquire beautiful images of very dim fluorescence specimens, fast enough to record the most dynamic processes, have high resolution to capture the finest detail, and have enough useful dynamic range to accurately measure minute differences in intensity. In fluorescence microscopy, cellular components are labeled with molecules that emit photons when illuminated with excitatory light of the appropriate wavelength. Acquiring fluorescence images presents a special challenge because of the relatively low number of photons emitted by the specimen that must be collected by the microscope optics and detected by the camera. An image acquisition system minimally consists of a microscope and a camera, but may also include a computer, imaging software, and a variety of motorized components for automated acquisition.