Chapter 9 High-Resolution Video-Enhanced Differential Interference Contrast (VE-DIC) Light Microscopy

Abstract

Publisher Summary Differential interference contrast (DIC) light microscopy was an immediate success after its introduction in the 1960s because it could produce high-contrast optical images of the edges of objects and fine-structural detail within transparent specimens. DIC contrast depends on gradients in optical path and not on absolute values of optical path (OP). As a result, DIC methods can produce clear optical sections of relatively thick transparent specimens. Resolution in DIC microscopy is also superior to phase contrast. VE–DIC has been important in many aspects of cell biology, including measuring the assembly dynamics of individual microtubules and forced production by microtubule assembly/disassembly. The quality of images in VE–DIC microscopy depends critically on both the quality of the image projected onto the camera detector by the microscope as well as on the electronic contrast enhancement capabilities of the camera. The chapter describes the basic concepts of DIC image formation and analog and digital contrast enhancement, the practical aspects of component selection, alignment and specimen chambers for VE–DIC, and several test specimens for measuring microscope and video performance.