High-speed scattering polarimetry for correlative nerve fiber imaging and multi-modal analysis.

Three-Dimensional Polarized Light Imaging (3D-PLI) and Computational Scattered Light Imaging (ComSLI) map dense nerve fibers in brain sections with micrometer resolution using visible light. 3D-PLI reconstructs single fiber orientations, while ComSLI captures multiple directions per pixel, offering deep insights into brain tissue structure. Here, we introduce the Scattering Polarimeter, a high-speed correlative microscope to leverage the strengths of both methods. Based on a Müller polarimeter, it incorporates variable retarders and a large-area light source for direct and oblique illumination, enabling rapid 3D-PLI and ComSLI measurements as well as measuring the Müller matrix per pixel. Applied to human and vervet monkey brain sections, the Scattering Polarimeter generates results comparable to state-of-the-art 3D-PLI and ComSLI setups and creates a multi-modal fiber direction map, integrating the robust fiber orientations obtained from 3D-PLI with fiber crossings from ComSLI. Furthermore, we discuss applications of the Scattering Polarimeter for unprecedented correlative and multi-modal brain imaging.