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Correlative light and volume electron microscopy to study brain development.

Shuichi Hayashi1, Nobuhiko Ohno2,3, Graham Knott4

  • 1Department of Anatomy, Kawasaki Medical School, 577 Matsushima, Kurashiki, Okayama 701-0192, Japan.

Microscopy (Oxford, England)
|January 9, 2023
PubMed
Summary
This summary is machine-generated.

Correlative light and volume electron microscopy (vCLEM) advances brain architecture understanding by linking 3D ultrastructure with physiological data. This technique, enhanced by genetic tools, aids in studying brain development and gene function.

Keywords:
brain developmentcerebral cortexcorrelative light and electron microscopy (CLEM)serial block-face scanning electron microscope (SBF-SEM)thalamusvolume electron microscopy

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Area of Science:

  • Neuroscience
  • Cell Biology
  • Microscopy

Background:

  • Volume electron microscopy (EM) provides high-resolution 3D ultrastructural details of brain architecture.
  • Correlating light microscopy (LM) with EM data is crucial for linking cellular function with structure.
  • Understanding brain development requires advanced imaging techniques to visualize complex neural circuits.

Approach:

  • Correlative light and volume electron microscopy (vCLEM) integrates LM and EM for multi-modal brain analysis.
  • Immunostaining-free vCLEM utilizes natural landmarks and genetic tools (fluorescent proteins, soybean ascorbate peroxidase) for seamless correlation.
  • vCLEM is applied to in vivo two-photon Ca2+ imaging and synaptic connection studies in thalamic neurons.

Key Points:

  • vCLEM enables the study of developmental processes in the brain by combining ultrastructural and physiological data.
  • Genetic manipulation tools, including CRISPR-Cas9, combined with vCLEM, facilitate protein localization and gene function studies.
  • Natural landmarks and genetic reporters streamline vCLEM, reducing the need for complex staining protocols.

Conclusions:

  • Volume EM and vCLEM significantly enhance the understanding of brain development and neural connectivity.
  • Future applications include advanced gene function analysis and protein localization studies in developmental neuroscience.
  • Combinatorial use of genetic tools with vCLEM promises deeper insights into the regulatory mechanisms of brain development.