Optical Clearing and Labeling for Light-sheet Fluorescence Microscopy in Large-scale Human Brain ImagingDanila Di Meo *1, Josephine Ramazzotti *1, Marina Scardigli *1,2, Franco Cheli 1, Luca Pesce 1,3, Niamh Brady 1, Giacomo Mazzamuto 1,4,5, Irene Costantini 1,4,6, Francesco S. Pavone 1,4,5
1European Laboratory for Non-linear Spectroscopy (LENS), University of Florence, 2Division of Physiology, Department of Experimental and Clinical Medicine, University of Florence, 3Department of Physics, University of Pisa, 4National Research Council - National Institute of Optics (CNR-INO), 5Department of Physics and Astronomy, University of Florence, 6Department of Biology, University of Florence
The present protocol provides a step-by-step procedure for rapid and simultaneous optical clearing, muti-round labeling, and 3D volumetric reconstruction of tens of postmortem human brain sections by combining the (SWITCH - H2O2 - Antigen Retrieval - 2,2'-thiodiethanol [TDE]) SHORT tissue transformation technique with light-sheet fluorescence microscopy imaging in a routinely high-throughput protocol.
