MAI-SIM: interferometric multicolor structured illumination microscopy for everybody

Abstract

Sub-diffraction resolution, gentle sample illumination, and the possibility to image in multiple colors make Structured Illumination Microscopy (SIM) an imaging technique which is particularly well suited for live cell observations. Here, we present Machine learning Assisted Interferometric-SIM (MAI-SIM), an easy-to-implement method for high speed SIM imaging in multiple colors. The instrument is based on an interferometer design in which illumination patterns are generated, rotated, and stepped in phase through movement of a single galvanometric mirror element. The design is robust, flexible, and the pattern generation process works for all wavelengths. We complement the unique properties of interferometric SIM with a machine learning toolbox that is simple and efficient to use and is superior to existing methods for the reconstruction of super-resolved images recorded by the instrument. The framework permits real-time SIM reconstructions to be performed in multiple colors, providing the user with instant visualization of the super-resolved images. We demonstrate the capability of MAI-SIM on live biological samples and capture super-resolution images in multiple colors simultaneously over large fields of view. Finally, we embrace a fully open design philosophy to bring the advantages of MAI-SIM to as many users as possible and provide full details on system design and software.