Scattering polarimetry enables correlative nerve fiber imaging and multimodal analysis
Franca auf der Heiden, Markus Axer, Katrin Amunts, Miriam Menzel
Abstract
Mapping the intricate network of nerve fibers is crucial for understanding brain function. 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 3D-fiber orientations, while ComSLI disentangles multiple directions per pixel. So far, these imaging techniques have been realized in separate setups. A combination within a single device would facilitate faster measurements, pixelwise mapping, cross-validation of fiber orientations, and leverage the advantages of each technique while mitigating their limitations. Here, we introduce the Scattering Polarimeter, a microscope that facilitates correlative large-area scans by integrating 3D-PLI and ComSLI measurements into a single system. Based on a Mueller polarimeter, it incorporates variable retarders and a large-area light source for direct and oblique illumination, enabling combined 3D-PLI and ComSLI measurements. 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 multimodal 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 multimodal brain imaging.