A Guide to Perform 3D Histology of Biological Tissues with Fluorescence Microscopy

Year: 2023

Authors: Laurino A., Franceschini A., Pesce L., Cinci L., Montalbano A., Mazzamuto G., Sancataldo G., Nesi G., Costantini I., Silvestri L., Pavone FS.

Autors Affiliation: LENS, European Lab Nonlinear Spect, I-50019 Sesto Fiorentino, Italy; Univ Florence, Dept Phys & Astron, I-50019 Florence, Italy; Careggi Univ Hosp, Dept Expt & Clin Biomed Sci, Radiodiagnost Unit n 2, I-50134 Florence, Italy; Univ Florence, Dept Neurofarba Sect Pharmacol & Toxicol, I-50139 Florence, Italy; Natl Inst Opt CNR INO, Natl Res Council, I-50125 Sesto Fiorentino, Italy; Univ Florence, Dept Hlth Sci, I-50139 Florence, Italy; Univ Florence, Dept Biol, I-50019 Florence, Italy; Univ Siena, Dept Mol & Dev Med, I-53100 Siena, Italy; Scuola Normale Super Pisa, NEST Lab, Piazza San Silvestro 12, I-56127 Pisa, Italy; Univ Florence, Dept Expt & Clin Med, I-50134 Florence, Italy; Univ Palermo, Dept Phys & Chem, I-90128 Palermo, Italy.

Abstract: The analysis of histological alterations in all types of tissue is of primary importance in pathology for highly accurate and robust diagnosis. Recent advances in tissue clearing and fluorescence microscopy made the study of the anatomy of biological tissue possible in three dimensions. The combination of these techniques with classical hematoxylin and eosin (H&E) staining has led to the birth of three-dimensional (3D) histology. Here, we present an overview of the state-of-the-art methods, highlighting the optimal combinations of different clearing methods and advanced fluorescence microscopy techniques for the investigation of all types of biological tissues. We employed fluorescence nuclear and eosin Y staining that enabled us to obtain hematoxylin and eosin pseudo-coloring comparable with the gold standard H&E analysis. The computational reconstructions obtained with 3D optical imaging can be analyzed by a pathologist without any specific training in volumetric microscopy, paving the way for new biomedical applications in clinical pathology.

Journal/Review: INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES

Volume: 24 (7)      Pages from: 6747-1  to: 6747-18

More Information: The research leading to these results received funding from the European Union’s Horizon 2020 Framework Programme for Research and Innovation under the Specific Grant Agreement No. 785907 (Human Brain Project SGA2), No. 945539 (Human Brain Project SGA3), and No. 871124 (Laserlab-Europe); the General Hospital Corporation Center of the National Institutes of Health under the award number U01 MH117023; the Italian Ministry for Education in the framework of the Euro-Bioimaging Italian Node (ESFRI research infrastructure); and Fondazione CR Firenze (private foundation, project title: Human Brain Optical Mapping). The content of this work is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
KeyWords: 3D histology; clearing methods; H&E staining; Light Sheet Microscopy; Two Photon Fluorescence Microscopy
DOI: 10.3390/ijms24076747

Citations: 5
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