0000000000123035
AUTHOR
Irene Costantini
Swift light sheet volumetric charting of large human brain portions
Using a custom light sheet fluorescence microscope, we image large stained human brain portions, labelled for NeuN and GAD67 neuronal markers, discerning the inhibitory population via neural-network based image analysis and exposing the brain connectivity.
A Guide to Perform 3D Histology of Biological Tissues with Fluorescence Microscopy
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 …
3D molecular phenotyping of cleared human brain tissues with light-sheet fluorescence microscopy
AbstractThe combination of optical tissue transparency with immunofluorescence allows the molecular characterization of biological tissues in 3D. However, adult human organs are particularly challenging to become transparent because of the autofluorescence contributions of aged tissues. To meet this challenge, we optimized SHORT (SWITCH—H2O2—antigen Retrieval—TDE), a procedure based on standard histological treatments in combination with a refined clearing procedure to clear and label portions of the human brain. 3D histological characterization with multiple molecules is performed on cleared samples with a combination of multi-colors and multi-rounds labeling. By performing fast 3D imaging…