Long Preservation of AAV-Transduced Fluorescence by a Modified Organic Solvent-Based Clearing Method

Tao Lu, Munehisa Shinozaki, Narihito Nagoshi, Masaya Nakamura, Hideyuki Okano

Research output: Contribution to journalArticlepeer-review


The development of tissue clearing technologies allows 3D imaging of whole tissues and organs, especially in studies of the central nervous system innervated throughout the body. Although the three-dimensional imaging of solvent-cleared organs (3DISCO) method provides a powerful clearing capacity and high transparency, the rapid quenching of endogenous fluorescence and peroxide removal process decreases its practicability. This study provides a modified method named tDISCO to solve these limitations. The tDISCO protocol can preserve AAV-transduced endogenous EGFP fluorescence for months and achieve high transparency in a fast and simple clearing process. In addition to the brain, tDISCO was applied to other organs and even hard bone tissue. tDISCO also enabled us to visualize the long projection neurons and axons with high resolution. This method provides a fast and simple clearing protocol for 3D visualization of the AAV- transduced long projection neurons throughout the brain and spinal cord.

Original languageEnglish
Article number9637
JournalInternational journal of molecular sciences
Issue number17
Publication statusPublished - 2022 Sept


  • AAV
  • endogenous fluorescence
  • light-sheet fluorescence microscopy
  • retrograde tracing
  • tissue clearing

ASJC Scopus subject areas

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry


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