The three-dimensional reconstruction of large volumes of the human neural networks at cellular resolution is one of thebiggest challenges of our days. Commonly, fine slices of samples marked with colorimetric techniques are individuallyimaged. This approach in addition to being time-consuming does not consider space cell organization, leading to loss ofinformation. The aim of this work was to develop a methodology that allows analyzing the cytoarchitecture of the humanbrain in three dimensions at high resolution. In particular, we exploit the possibility of combining high-resolution 3Dimaging techniques with clearing methodologies. We successfully integrate the SWITCH immunohistochemistrytechnique with the TDE clearing method to image a large volume of human brain tissue with two-photon fluorescencemicroscopy. In conclusion, this new approach enables to characterize large human brain specimens with high-resolutionoptical techniques, giving the possibility to expand the histological studies to the third dimension.
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