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Related Concept Videos

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Transmission electron microscopy (TEM) can be used to determine the 3D structure of biological samples with the help of techniques such as electron microscope tomography and single-particle reconstruction. While single-particle reconstruction can examine macromolecules and macromolecular complexes in vitro conditions only, tomography permits the study of cell components or small cells in vivo.
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Related Experiment Video

Updated: Sep 24, 2025

Targeted Labeling of Neurons in a Specific Functional Micro-domain of the Neocortex by Combining Intrinsic Signal and Two-photon Imaging
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Single-Neuron Labeling in Fixed Tissue and Targeted Volume Electron Microscopy.

Marta Turegano-Lopez1,2, Andrea Santuy3, Asta Kastanauskaite1

  • 1Laboratorio Cajal de Circuitos Corticales, Centro de Tecnología Biomédica, Universidad Politécnica de Madrid, Pozuelo de Alarcón, Spain.

Frontiers in Neuroanatomy
|May 9, 2022
PubMed
Summary

This study introduces a new method for 3D reconstruction of neural tissue, enabling detailed analysis of synaptic connections across different scales. The technique combines single-neuron labeling with advanced imaging for precise ultrastructural mapping.

Keywords:
3D reconstructionFIB-SEMintracellular injectionquantitative neuroanatomyserial sectioning

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Area of Science:

  • Neuroscience
  • Cell Biology
  • Microscopy

Background:

  • Nervous tissue complexity hinders understanding neural connectivity across scales.
  • Existing methods struggle to correlate light and electron microscopic data for single cells.
  • Bridging mesoscopic and nanoscopic scales in neural circuits is technically challenging.

Purpose of the Study:

  • To develop an effective method for 3D reconstruction of subcellular structures at the ultrastructural level.
  • To enable correlation of light microscopic labeling with electron microscopic synaptic analysis in fixed tissue.
  • To facilitate detailed mapping of single-neuron synaptic connectivity and neuropil analysis.

Main Methods:

  • Single-neuron labeling via intracellular injection of horseradish peroxidase (HRP) in fixed vibratome sections.
  • Flat-embedding, re-sectioning, and automated focused ion beam milling and scanning electron microscopy (FIB-SEM) imaging.
  • 3D reconstruction and analysis of high-resolution image series for ultrastructural detail.

Main Results:

  • Successful 3D reconstruction of labeled subcellular structures at the ultrastructural level.
  • Accurate correlation of light microscopic cellular segments with their synaptic connections at the electron microscopic level.
  • Facilitation of analysis of synaptic connectivity for individual neurons and surrounding neuropil.

Conclusions:

  • The presented method effectively bridges the gap between light and electron microscopy for neural circuit analysis.
  • This technique allows detailed investigation of synaptic organization within specific cellular compartments and brain regions.
  • It offers a powerful tool for understanding neural connectivity and ultrastructural features of neurons.