3D method via time and spatially multiplexed confocal microscope

A confocal microscope can achieve superior contrast when imaging a certain layer in the bulky samples. However, the parallel signal collection feature of the optical system is sacrificed when the sample is scanned pixel by pixel. In this paper, we proposed a novel confocal microscope design that uses a time and spatially multiplexed method, which dramatically increases the time resolution of a confocal microscope. This design has been used to solve a long-standing problem in cardiac research whether or not a small submembranous domain exists with calcium and sodium ion concentrations significantly different from those measured in bulk cytosol. We applied our time and spatially multiplexed confocal microscope to obtain the transient 3-D distribution of calcium ion concentration in rat cardiac myocytes. Our experimental results prove the feasibility of the technique and also demonstrate the huge potential of this design.