Miniature structured illumination microscope for in vivo 3D imaging of brain structures with optical sectioning
Omkar D. Supekar, Andrew Sias, Sean R. Hansen, Gabriel Martinez, Graham C. Peet, Xiaoyu Peng, Victor M. Bright, Ethan G. Hughes, Diego Restrepo, Douglas P. Shepherd, Cristin G. Welle, Juliet T. Gopinath, Emily A. Gibson
Abstract
We present a high-resolution miniature, light-weight fluorescence microscope with electrowetting lens and onboard CMOS for high resolution volumetric imaging and structured illumination for rejection of out-of-focus and scattered light. The miniature microscope (SIMscope3D) delivers structured light using a coherent fiber bundle to obtain optical sectioning with an axial resolution of 18 µm. Volumetric imaging of eGFP labeled cells in fixed mouse brain tissue at depths up to 260 µm is demonstrated. The functionality of SIMscope3D to provide background free 3D imaging is shown by recording time series of microglia dynamics in awake mice at depths up to 120 µm in the brain.