What is the significance of dendritic spines?

Dendritic spines are crucial for synaptic connections and play a key role in neuronal communication.

How does focused ion beam scanning electron microscopy work?

It uses an ion beam to remove layers of tissue, allowing for detailed imaging of the exposed surfaces.

What are the benefits of using osmium for staining?

Osmium binds to lipids and enhances contrast, making it easier to visualize cellular structures.

Can this method be applied to other types of tissues?

Yes, the techniques can be adapted for imaging various types of biological tissues.

What are the limitations of this imaging technique?

The process can be time-consuming and requires specialized equipment and expertise.

Focused Ion Beam Scanning Electron Microscopy for 3D Reconstruction of Synaptically Linked Neurons

JoVE Encyclopedia of Experiments

Neuroscience

0 views • 1:21 min • May 30th, 2025

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Take a fixed section of mouse cortical brain tissue exhibiting neuronal axons and dendrites. The dendrites possess protrusions, known as dendritic spines, which form synapses with axons.

Incubate the tissue with osmium-ferrocyanide, which binds to lipids, followed by another osmium application, which enhances structural contrast for imaging.

Stain with a heavy metal compound that binds to macromolecules and enhances contrast.

Dehydrate using increasing alcohol concentrations for resin embedding.

Embed the tissue in resin, mount it on a holder, and coat it with gold to optimize imaging quality.

Use focused ion beam scanning electron microscopy for three-dimensional imaging.

Employ an ion beam to remove a thin layer of resin-embedded tissue, exposing an imaging surface.

Focus an electron beam on the exposed surface and detect the backscattered electrons to generate an image.

Perform serial slicing and imaging and reconstruct a three-dimensional image, revealing a synaptically linked cortical neuronal network.