Practical design criteria for a dynamic ratio imaging system

Problems encountered and solutions devised during the construction of a productive Ca2(+)-imaging system are discussed. Many of these relate to the rapid and interactive nature of experiments on cytosolic Ca2+. The emphasis on accurate photometric quantitation of dynamically changing images contrasts with that of most image-processing software packages, which concentrate either on leisurely massage of static images or on descriptions of lateral motions of objects without concern for their brightnesses. Particularly important goals for Ca2(+)-imaging include real-time ratioing, psychophysically effective display formats, ease of experiment annotation, mass storage of image sequences, automated extraction of time courses and population statistics, aids to presenting images for seminars and publications, and program modifiability.