Optical sectioning in differential interference contrast microscopy

While developing a three-dimensional (3-D) measurement technique for the retardation-modulated differential interference contrast (RM-DIC) microscope, we encountered a problem. The problem was that the measurement range was restricted in λ because it applies weak phase approximation.To overcome this drawback, we propose a 3-D reconstruction method with z-axis scanning. This technique needs high optical sectioning, like confocal microscopy.We investigated the characteristic of optical sectioning in a DIC microscope, then we confirmed experimentally that a DIC microscope has high optical sectioning. We also confirmed that a RM-DIC microscope has higher optical sectioning.By combining the optical sectioning of a RM-DIC microscope and z-scanning, we developed a new 3-D reconstruction method. This novel technique overcomes the observed problem as the measurement range is increased to micron order.