Real-time registration and display of confocal microscope imagery for multiple-band analysis

Multiple images acquired in real-time from a confocal microscope in different illumination wavebands are used to construct a three-dimensional understanding of living cell signal propagation. Registration, fusion, and display of these images are performed in real-time using field programmable gate array (FPGA) technology to allow the biological scientist to observe time critical signaling processes as they occur. The registration and display system operates at a rate of up to 15 frames/s, and performs dark offset correction and noise reduction, followed by non-linear correction for lens warp and focal plane tilt, and finally correction for spatial mismatch between the different wavebands. Parameters for the registration process are determined automatically from calibration images at the start of each experimental session in real-time. The resulting fused images exhibit significantly enhanced detail as illustrated by examples presented in the paper.