Three-beam interference with circular polarization for structured illumination microscopy

Abstract

Three-dimensional structured illumination microscopy (3D-SIM) is a wide-field technique that can provide doubled resolution and improved image contrast. In this work, we demonstrate a simple approach to 3D-SIM - using three-beam interference with circular polarization to generate the pattern of structured illumination, so that the modulation contrast is routinely maintained at all orientations without a complicated polarization rotator or mechanical motion. We derive the resultant intensity distribution of the interference pattern to confirm the modulation contrast independent of orientation, and compare the result with those using interfering beams of linear polarization. To evaluate the influence of the modulation contrast on imaging, we compare the simulated SIM images of 100-nm beads. Experimental results are presented to confirm the simulations. Our approach requires merely a λ/4-wave plate to alter the interfering beams from linear to circular polarization. This simplicity together with the use of a spatial light modulator to control the interference pattern facilitates the implementation of a 3D-SIM system and should broaden its application.