DEVELOPMENT OF NEW AUTOMATED POLARIZED LIGHT MICROSCOPE

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The polarized light microscope has the unique potential to measure submicroscopic molecular arrangements dynamically and non-destructively in living cells and other specimens. With the traditional pol-scope, however, single images display only those anisotropic structures that have a limited range of orientations with respect to the polarization axes of the microscope. Furthermore, rapid measurements are restricted to a single image point or single area that exhibits uniform birefringence or other form of optical anisotropy, while measurements comparing several image points take an inordinately long time. We propose to develop a new kind of polarized light microscope which will combine speed and high resolution in its measurement of the specimen anisotropy, irrespective of its orientation. The design of the new pol-scope is based on the traditional polarized light microscope with two essential modifications: circular polarizers will replace linear polarizers and two electro-optical modulators will replace the traditional compensator. A video camera and computer assisted image analysis will provide measurements of specimen anisotropy once every second for all points of the image comprising the field of view. The measurement process will result in particularly clear images free of shading and background light over the entire viewing field. The images and measurements will document fine structural and molecular organization within a thin optical section of the specimen. The high spatial and temporal resolution of the new pol-scope together with its' ease of use for obtaining quantitative specimen records, concurrently for the whole field of view, will allow new investigations into the dynamics of macromolecular organization of living cells and other systems.