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Optical clearing of fixed brain samples using SeeDB.

Meng-Tsen Ke1, Takeshi Imai

  • 1Laboratory for Sensory Circuit Formation, RIKEN Center for Developmental Biology, Kobe, Japan; Graduate School of Biostudies, Kyoto University, Kyoto, Japan.

Current Protocols in Neuroscience
|February 11, 2014
PubMed
Summary
This summary is machine-generated.

We developed SeeDB, a novel optical clearing agent for brain tissue. This method enables deep-tissue fluorescence imaging of neuronal circuitry with preserved morphology.

Keywords:
fluorescent imagingoptical clearingtwo-photon microscopy

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Area of Science:

  • Neuroscience
  • Biotechnology
  • Microscopy

Background:

  • Understanding neuronal circuitry requires advanced imaging techniques.
  • Current methods often struggle with deep tissue penetration and dye preservation.

Purpose of the Study:

  • To introduce SeeDB, a water-based optical clearing agent.
  • To demonstrate its efficacy in preserving fluorescent signals and cellular morphology.
  • To enable large-scale 3D imaging of neural networks.

Main Methods:

  • Developed a novel water-based optical clearing agent, SeeDB.
  • Applied SeeDB to fixed brain samples.
  • Utilized confocal and two-photon microscopy for imaging.

Main Results:

  • SeeDB effectively clears fixed brain samples within days.
  • It preserves a wide range of fluorescent dyes, including fluorescent proteins and lipophilic tracers.
  • Maintains constant sample volume, preserving cellular morphology.
  • Achieved imaging depths exceeding 1000 µm with confocal microscopy.
  • Enabled millimeter-scale imaging of fixed mouse brains using two-photon microscopy.

Conclusions:

  • SeeDB is a robust method for optical clearing of brain tissue.
  • Facilitates deep-tissue, high-resolution 3D fluorescence imaging of neuronal structures.
  • Significantly advances the study of complex neural circuits.