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Whole-brain block-face serial microscopy tomography at subcellular resolution using FAST.

Kaoru Seiriki1,2, Atsushi Kasai3, Takanobu Nakazawa1,4

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We developed a fast, high-resolution imaging technique called block-face serial microscopy tomography (FAST) for whole mouse brains. This method allows detailed analysis of neural structures and cell types for brain system research.

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Area of Science:

  • Neuroscience
  • Microscopy
  • Bioimaging

Background:

  • High-speed imaging of cleared brains is limited by spatial resolution.
  • Conventional confocal microscopy offers high resolution but is slow.
  • A method is needed to combine speed and high resolution for brain imaging.

Purpose of the Study:

  • To describe an optimized protocol for block-face serial microscopy tomography (FAST).
  • To enable high-speed, axonal-resolution fluorescence imaging of fixed brains.
  • To facilitate brain-wide anatomical and functional analyses.

Main Methods:

  • Detailed protocol for FAST hardware assembly, sample preparation, and imaging.
  • Serial section fluorescence imaging of fixed and cleared brains.
  • Image data processing for analysis.

Main Results:

  • FAST achieves axonal spatial resolution in fluorescence imaging.
  • A single mouse brain can be imaged in as little as 2.4 hours.
  • Sample preparation is adaptable, allowing simultaneous processing of multiple samples.

Conclusions:

  • FAST overcomes limitations of existing microscopy techniques for brain imaging.
  • The protocol enables detailed structural and cell-type-specific brain analyses.
  • FAST supports unbiased, hypothesis-free research in systems neuroscience.