page_title: Differential interference contrast microscopy
section_title: Image
hierarchical_section_title: Differential interference contrast microscopy / Image
caption_attribution_description: Example of image construction by en:differential interference contrast microscopy.
context_page_description: Differential interference contrast microscopy, also known as Nomarski interference contrast or Nomarski microscopy, is an optical microscopy technique used to enhance the contrast in unstained, transparent samples. DIC works on the principle of interferometry to gain information about the optical path length of the sample, to see otherwise invisible features. A relatively complex optical system produces an image with the object appearing black to white on a grey background. This image is similar to that obtained by phase contrast microscopy but without the bright diffraction halo. The technique was developed by Polish physicist Georges Nomarski in 1952.
  DIC works by separating a polarized light source into two orthogonally polarized mutually coherent parts which are spatially displaced at the sample plane, and recombined before observation. The interference of the two parts at recombination is sensitive to their optical path difference.
context_section_description: The image has the appearance of a three-dimensional object under very oblique illumination, causing strong light and dark shadows on the corresponding faces. The direction of apparent illumination is defined by the orientation of the Wollaston prisms.
  As explained above, the image is generated from two identical bright field images being overlaid slightly offset from each other (typically around 0.2 μm), and the subsequent interference due to phase difference converting changes in phase (and so optical path length) to a visible change in darkness. This interference may be either constructive or destructive, giving rise to the characteristic appearance of three dimensions.
  The typical phase difference giving rise to the interference is very small, very rarely being larger than 90° (a quarter of the wavelength). This is due to the similarity of refractive index of most samples and the media they are in: for example, a cell in water only has a refractive index difference of around 0.05. This small phase difference is important for the correct function of DIC, since if the phase difference at the joint between two substances is too large then the phase difference could reach 180° (half a wavelength), resulting in complete destructive interference and an anomalous dark region; if the phase difference reached 360° (a full wavelength), it would produce complete constructive interference, creating an anomalous bright region.
  The image can be approximated (neglecting refraction and absorption due to the sample and the resolution limit of beam separation) as the differential of optical path length with respect to position across the sample along the shear, and so the differential of the refractive index (optical density) of the sample.
  The contrast can be adjusted using the offset phase, either by translating the objective Nomarski prism, or by a lambda/4 waveplate between polarizer and the condenser Normarski prism (De-Senarmont Compensation). The resulting contrast is going from dark-field for zero phase offset (intensity proportional to the square of the shear differential), to the typical relief seen for phase of ~5–90 degrees, to optical staining at 360 degrees, where the extinguished wavelength shifts with the phase differential.
caption: The process of image production in a DIC microscope
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