A differential interference contrast (DIC)
microscope system is provided comprising: (a) an illumination source for illuminating a sample ; (b) a lens
system for viewing the illuminated sample, including an objective, defining an
optical axis; (c) at least one
detector system for receiving a
sample image; (d) mechanisms for
wavelength multiplexing the shear direction or shear magnitude or both on the sample and demultiplexing the
resultant DIC images on the
detector; and (e) a mechanism for modulating the phase of the interference image. Various approaches are disclosed to accomplish
wavelength multiplexing of shear direction and demultiplexing the two DIC images that result. It is possible for the two,
wavelength multiplexed DIC images to differ in either or both shear direction or magnitude. These approaches include (1) two DIC microscopes, each operating at a different wavelength, but which share a single objective through a
beam splitter; (2) a segmented DIC
prism that is made in four sections where opposite sections are paired and have the same shear direction and amount, and each pair of sections have filters transmitting different wavelengths; (3) a segmented DIC
prism that is located in or near an aperture stop or
pupil of said DIC
microscope to obtain data in two shear directions that is multiplexed by wavelength; (4) a dual field-of-view optical system with two DIC prisms, one in each path to wavelength
multiplex shear direction or shear magnitude through said objective; (5) demultiplexing wavelength multiplexed DIC images through the use of a wavelength selective
beam splitter and two detectors; (6) demultiplexing wavelength multiplexed DIC images through the use of a wavelength
controlled source and a single
detector; and (7) demultiplexing wavelength multiplexed DIC images through the use of dual field-of-view
optics and a single detector. These various approaches permit rapid, robust measurement of slope in two directions. Further, phase shifting and DIC
microscopy are limited to measurements within the
depth of focus (DOF) of the objective while WLI
microscopy is not.