Efficient 3D light-sheet imaging of very large-scale optically cleared human brain and prostate tissue samples
Anna Schueth, Sven Hildebrand, Iryna Samarska, Shubharthi Sengupta, Annemarie Kiessling, Andreas Herrler, Axel zur Hausen, Michael Capalbo, Alard Roebroeck
Abstract
Abstract 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 cm 3 sized formalin fixed human brain and up to ~7 cm 3 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 cm 3 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/cm 3 ), (b) Mosaic 4 (4.1 µm isotropic resolution, ~ 5 h/cm 3 ) and (c) Mosaic 0.5 (0.5 µm near isotropic resolution, ~15.8 h/cm 3 ). 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.