Clear cell DIC observation

Clear cell DIC observation.


Phase contrast microscopy produces image intensity (amplitude) values that vary as a function of specimen optical path length magnitude, with very dense regions (those having large path lengths) appearing darker than the background. Alternatively, specimen features that have relatively low thickness values, or a refractive index less than the surrounding medium, are rendered much lighter when superimposed on the standard (positive) phase contrast medium gray background.


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Phase contrast microscopy produces image intensity (amplitude) values that vary as a function of specimen optical path length magnitude, with very dense regions (those having large path lengths) appearing darker than the background. Alternatively, specimen features that have relatively low thickness values, or a refractive index less than the surrounding medium, are rendered much lighter when superimposed on the standard (positive) phase contrast medium gray background.



The situation is quite dissimilar for differential interference contrast (DIC) microscope A19.0204, where optical path length gradients (in effect, the rate of change in the direction of wavefront shear) are primarily responsible for contrast. Steep gradients in path length generate excellent contrast, and images display the pseudo three-dimensional relief shading, which is characteristic of the DIC technique. Regions having very shallow optical path slopes, such as those observed in extended, flat specimens, produce insignificant contrast and often appear in the image at the same intensity level as the background.


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