Three-dimensional imaging of solvent-cleared organs using 3DISCO.
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.
Department of Neuroscience, Genentech, South San Francisco, California, USA. erturk.ali@gene.com
Ali
A
Ertürk
Klaus
K
Becker
Nina
N
Jährling
Christoph P
CP
Mauch
Caroline D
CD
Hojer
Jackson G
JG
Egen
Farida
F
Hellal
Frank
F
Bradke
Morgan
M
Sheng
Hans-Ulrich
HU
Dodt
D000818:Animals; D001921:Brain / Q000033:anatomy & histology* / Q000098:blood supply; D005663:Furans / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000737:chemistry; D006207:Half-Life; D021621:Imaging, Three-Dimensional / Q000379:methods*; D051379:Mice; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence / Q000379:methods*; D010647:Phenyl Ethers / Q000737:chemistry*; D012997:Solvents / Q000737:chemistry*; D013116:Spinal Cord / Q000033:anatomy & histology* / Q000098:blood supply
D005663:Furans; D010647:Phenyl Ethers; D012997:Solvents; D049452:Green Fluorescent Proteins; C076624:dibenzyl ether; C018674:tetrahydrofuran