biostudies-literatureUnknown6(1)Schueth AThe ability to image human tissue samples in 3D, with both cellular resolution and a large field of view (FOV), can improve fundamental and clinical investigations. Here, we demonstrate the feasibility of light-sheet imaging of ~5 cm³ sized formalin fixed human brain and up to ~7 cm³ sized formalin fixed paraffin embedded (FFPE) prostate cancer samples, processed with the FFPE-MASH protocol. We present a light-sheet microscopy prototype, the cleared-tissue dual view Selective Plane Illumination Microscope (ct-dSPIM), capable of fast 3D high-resolution acquisitions of cm³ scale cleared tissue. We used mosaic scans for fast 3D overviews of entire tissue samples or higher resolution overviews of large ROIs with various speeds: (a) Mosaic 16 (16.4 µm isotropic resolution, ~1.7 h/cm³), (b) Mosaic 4 (4.1 µm isotropic resolution, ~ 5 h/cm³) and (c) Mosaic 0.5 (0.5 µm near isotropic resolution, ~15.8 h/cm³). We could visualise cortical layers and neurons around the border of human brain areas V1&V2, and could demonstrate suitable imaging quality for Gleason score grading in thick prostate cancer samples. We show that ct-dSPIM imaging is an excellent technique to quantitatively assess entire MASH prepared large-scale human tissue samples in 3D, with considerable future clinical potential.Communications biology170https://www.ebi.ac.uk/biostudies/studies/S-EPMC9925784biostudies-literatureEfficient 3D light-sheet imaging of very large-scale optically cleared human brain and prostate tissue samples.PMC9925784Zur Hausen ASchueth AKiessling AHildebrand SSamarska IHerrler ASengupta SCapalbo MRoebroeck AfalseEfficient 3D light-sheet imaging of very large-scale optically cleared human brain and prostate tissue samples.The ability to image human tissue samples in 3D, with both cellular resolution and a large field of view (FOV), can improve fundamental and clinical investigations. Here, we demonstrate the feasibility of light-sheet imaging of ~5 cm³ sized formalin fixed human brain and up to ~7 cm³ sized formalin fixed paraffin embedded (FFPE) prostate cancer samples, processed with the FFPE-MASH protocol. We present a light-sheet microscopy prototype, the cleared-tissue dual view Selective Plane Illumination Microscope (ct-dSPIM), capable of fast 3D high-resolution acquisitions of cm³ scale cleared tissue. We used mosaic scans for fast 3D overviews of entire tissue samples or higher resolution overviews of large ROIs with various speeds: (a) Mosaic 16 (16.4 µm isotropic resolution, ~1.7 h/cm³), (b) Mosaic 4 (4.1 µm isotropic resolution, ~ 5 h/cm³) and (c) Mosaic 0.5 (0.5 µm near isotropic resolution, ~15.8 h/cm³). We could visualise cortical layers and neurons around the border of human brain areas V1&V2, and could demonstrate suitable imaging quality for Gleason score grading in thick prostate cancer samples. We show that ct-dSPIM imaging is an excellent technique to quantitatively assess entire MASH prepared large-scale human tissue samples in 3D, with considerable future clinical potential.2023-01-01T00:00:00Z2023 Feb2025-04-18T21:32:11.219Z2025-04-07T09:24:45.677ZS-EPMC99257843678193910.1038/s42003-023-04536-4