6533b870fe1ef96bd12d05a3
RESEARCH PRODUCT
3D molecular phenotyping of cleared human brain tissues with light-sheet fluorescence microscopy
Luca PesceMarina ScardigliVladislav GavryusevAnnunziatina LaurinoGiacomo MazzamutoNiamh BradyGiuseppe SancataldoLudovico SilvestriChristophe DestrieuxPatrick R. HofIrene CostantiniFrancesco S. Pavonesubject
AdultImaging Three-DimensionalMicroscopy FluorescenceBrainFluorescent Antibody TechniqueHumansMedicine (miscellaneous)Hydrogen PeroxideGeneral Agricultural and Biological SciencesSettore FIS/07 - Fisica Applicata(Beni Culturali Ambientali Biol.e Medicin)General Biochemistry Genetics and Molecular BiologyAgeddescription
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 of the samples with a custom-made inverted light-sheet fluorescence microscope (LSFM), we reveal fine details of intact human brain slabs at subcellular resolution. Overall, we proposed a scalable and versatile technology that in combination with LSFM allows mapping the cellular and molecular architecture of the human brain, paving the way to reconstruct the entire organ.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2022-05-12 | Communications Biology |