X-Clarity for automated tissue clearing and light-sheet microscopy in neurooncology and neuroimmunology
Establishing a pipeline for improved sample preparation and tissue clearing
In the ClearBrainTumor project, we established a pipeline to study primary or secondary tumors using light-sheet microscopy. In collaboration with existing infrastructure and expertise of the DKFZ Light Microscopy Core Facility (LMCF), we set out to improve the sample preparation and tissue clearing of the mouse brain tissues by introducing the X-CLARITY™ (Logos Biosystems Inc.) clearing apparatus. However, we successfully managed to troubleshoot and optimize working with the X-CLARITY on brain tumor samples, and image immune cell infiltration in melanoma brain metastases with light-sheet microscopy. The data was processed using the Zeiss Arivis Pro 3D Image Analysis (acquired with support from the Health and Life Science Alliance Explore!Tech funded project ViLTiS, 2023). Using this pipeline, we hope to better understand how T cells reach brain metastases and glioblastoma, and how we could improve efficacy of immune checkpoint inhibition therapies to treat these devastating diseases.
Light-sheet microscopy of a cleared sample showing T cells (red) infiltrating a melanoma brain tumor (green). By M. Shakouri, C. Thommek, M. Karreman and F. Bestvater.