Three-dimensional imaging of solvent-cleared organs using 3DISCO.

Autor: Ertürk A; Department of Neuroscience, Genentech, South San Francisco, California, USA. erturk.ali@gene.com, Becker K, Jährling N, Mauch CP, Hojer CD, Egen JG, Hellal F, Bradke F, Sheng M, Dodt HU
Jazyk: angličtina
Zdroj: Nature protocols [Nat Protoc] 2012 Nov; Vol. 7 (11), pp. 1983-95. Date of Electronic Publication: 2012 Oct 11.
DOI: 10.1038/nprot.2012.119
Abstrakt: The examination of tissue histology by light microscopy is a fundamental tool for investigating the structure and function of organs under normal and disease states. Many current techniques for tissue sectioning, imaging and analysis are time-consuming, and they present major limitations for 3D tissue reconstruction. The introduction of methods to achieve the optical clearing and subsequent light-sheet laser scanning of entire transparent organs without sectioning represents a major advance in the field. We recently developed a highly reproducible and versatile clearing procedure called 3D imaging of solvent-cleared organs, or 3DISCO, which is applicable to diverse tissues including brain, spinal cord, immune organs and tumors. Here we describe a detailed protocol for performing 3DISCO and present its application to various microscopy techniques, including example results from various mouse tissues. The tissue clearing takes as little as 3 h, and imaging can be completed in ∼45 min. 3DISCO is a powerful technique that offers 3D histological views of tissues in a fraction of the time and labor required to complete standard histology studies.
Databáze: MEDLINE