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Key Words: confocal, fluorescence, brightfield, phase contrast; Wollaston prism, de Sénarmont compensator, polarizing microscopy

Definition:DIC is a phase contrast technique that produces images with a characteristic '3-D' relief optical effect


DIC is a phase contrast technique that allows structure in transparent to be visualized by exploiting changes in refractive index. In contrast to conventional phase contrast techniques, which are based on changes in optical path length, DIC is based on the gradient of the optical path length (rate of change in wavefront shear). Steep gradients produce the high contrast and 3-D relief effect that is characteristic of DIC.


Contrast in DIC imaging is produced solely through an optical effect. This technique, therefore, is ideal for unstained living specimens (for example, cultured cells, embryos, blood smears, diatoms, protozoa). DIC provides morphological information without the need to use potentially toxic dyes and fluorophores. DIC is, however, often used in association with fluorescence to reveal morphological features of the specimen. It is also used for the examination of polymers and other materials.


DIC imaging requires several optical components; a polarizer, prism and specialized condenser. The correct set up and alignment of these components is central to successful DIC imaging. DIC capability can be installed on almost any brightfield transmitted, reflected, upright or inverted microscope.


Nikon's Eclipse 90i and 80i upright microscopes, TE2000 inverted series, and LV series have DIC capability. EZC1 software used in Nikon's EC1, C1, and C1si confocal systems enables easy switching between DIC and fluorescence imaging modes. Nikon's LiveScan Sweptfield system running on NIS Elements software also enables switching between DIC and fluorescence.


Differential Interference Contrast (DIC) Microscopy

DIC Microscope Component Alignment

Comparison of phase contrast and DIC

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