index n of the liquid can then be calculated from the maximum transmission angle θ as , where nG is the refractive index of the prism. This type of device is commonly used in chemical laboratories for identification of substances and for quality control. Handheld variants are used in agriculture by, e.g., wine makers to determine sugar content in grape juice, and inline process refractometers are used in, e.g., chemical and pharmaceutical industry for process control. In gemology, a different type of refractometer is used to measure the index of refraction and birefringence of gemstones. The gem is placed on a high refractive index prism and illuminated from below. A high refractive index contact liquid is used to achieve optical contact between the gem and the prism. At small incidence angles most of the light will be transmitted into the gem, but at high angles total internal reflection will occur in the prism. The critical angle is normally measured by looking through a telescope. Refractive index variations Unstained biological structures appear mostly transparent under Bright-field microscopy as most cellular structures do not attenuate appreciable quantities of light. Nevertheless, the variation in the materials that constitute these structures also corresponds to a variation in the refractive index. The following techniques convert such variation into measurable amplitude differences: To measure the spatial variation of the refractive index in a sample phase-contrast imaging methods are used. These methods measure the variations in phase of the light wave exiting the sample. The phase is proportional to the optical path length the light ray has traversed, and thus gives a measure of the integral of the refractive index along the ray path. The phase cannot be measured directly at optical or higher frequencies, and therefore needs to be converted into intensity by interference with a reference beam. In the visual spectrum this is done using Zernike phase-contrast microscopy, differential interference contrast microscopy (DIC), or interferometry. Zernike phase-contrast microscopy introduces a phase shift to the low spatial frequency components of the image with a phase-shifting annulus in the Fourier plane of the sample, so that high-spatial-frequency parts of the image can interfere with the low-frequency reference beam. In DIC the illumination is split up into two beams that are given different polarizations, are phase shifted differently, and are shifted transversely with slightly different amounts. After the specimen, the two parts are made to interfere, giving an