We report a method for extracting the birefringence properties of biological samples with micrometer-scale resolution in three dimensions, using a new form of polarization-sensitive optical coherence tomography. The method measures net retardance, net fast axis, and total reflectivity as a function of depth into the sample. Polarization sensing is accomplished by illumination of the sample with at least three separate polarization states during consecutive acquisitions of the same pixel, A scan, or B scan. The method can be implemented by use of non-polarization-maintaining fiber and a single detector. In a calibration test of the system, net retardance was measured with an average error of 7.5° (standard deviation 2.2°) over the retardance range 0° to 180°, and a fast axis with average error of 4.8° over the range 0° to 180°.
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