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

Nat Protoc. 2012 Nov;7(11):1983-95. doi: 10.1038/nprot.2012.119. Epub 2012 Oct 11.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / anatomy & histology*
  • Brain / blood supply
  • Furans / chemistry*
  • Green Fluorescent Proteins / analysis
  • Green Fluorescent Proteins / chemistry
  • Half-Life
  • Imaging, Three-Dimensional / methods*
  • Mice
  • Mice, Transgenic
  • Microscopy, Confocal
  • Microscopy, Fluorescence / methods*
  • Phenyl Ethers / chemistry*
  • Solvents / chemistry*
  • Spinal Cord / anatomy & histology*
  • Spinal Cord / blood supply


  • Furans
  • Phenyl Ethers
  • Solvents
  • Green Fluorescent Proteins
  • dibenzyl ether
  • tetrahydrofuran