We are developing an ultra-high-sensitivity and ultra-high-speed imaging system for bioscience, mainly for imaging of microbes with visible light and cells with fluorescence emission. Scarcity of photons is the most serious problem in applications of high-speed imaging to the scientific field. To overcome the problem, the system integrates new technologies consisting of (1) an ultra-high-speed video camera with sub-ten-photon sensitivity with the frame rate of more than 1 mega frames per second, (2) a microscope with highly efficient use of light applicable to various unstained and fluorescence cell observations, and (3) very powerful long-pulse-strobe Xenon lights and lasers for microscopes. Various auxiliary technologies to support utilization of the system are also being developed. One example of them is an efficient video trigger system, which detects a weak signal of a sudden change in a frame under ultra-high-speed imaging by canceling high-frequency fluctuation of illumination light. This paper outlines the system with its preliminary evaluation results.
Although we have aspired to observe dynamic changes in fluorescent images at the cellular level for a long time, the commercially available video cameras are not at all suitable for this purpose because of their low frame rates and photosensitivity. The present work tackles this issue and describes our attempt to find a solution by using our high-speed video camera and an ultrabright illumination system. We used light sources with considerably higher energy because conventional mercury lamps cannot produce sufficient brightness for our video cameras working a rate of more than 4,500 fps to obtain fluorescent images of cells. We observed that the flagellar movement of mice sperms ceased and multiple kinks developed in their tails when exposed to 2.7W of laser illumination for 1 s. In contrast, no significant alterations could be detected when the sperms were subjected to the same amount of energy by intermittent illumination. Since we found that cells can survive short-duration exposure to high-energy light, we attempted to construct an ultrabright Xenon-strobe illumination system. Our fluorescence studies are currently being extended to other types of animal cells, e.g., observation of the conduction of action potentials in the peripheral nerves of frog.