<em>In vivo</em> Imaging of the Mouse Spinal Cord Using Two-photon Microscopy

Dimitrios Davalos; Katerina Akassoglou

doi:10.3791/2760

Dans l'imagerie in vivo de la moelle épinière de souris utilisant microscop...

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JoVE Journal Neuroscience

In vivo Imaging of the Mouse Spinal Cord Using Two-photon Microscopy

Dans l'imagerie in vivo de la moelle épinière de souris utilisant microscopie à deux photons

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10:24 min

January 5, 2012

DOI: 10.3791/2760-v

Dimitrios Davalos¹, Katerina Akassoglou^1,2

¹Gladstone Institute of Neurological Disease,University of California, San Francisco , ²Department of Neurology,University of California, San Francisco

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Overview

This article describes a minimally invasive protocol for stabilizing the mouse spinal column and conducting repetitive in vivo spinal cord imaging using two-photon microscopy. The method aims to minimize respiratory-induced movements through a specific anesthetic regimen.

Key Study Components

Area of Science

Neuroscience
Imaging Techniques
Spinal Cord Research

Background

Stable in vivo imaging is crucial for observing cellular dynamics.
Two-photon microscopy allows for detailed imaging of living tissues.
Respiratory movements can interfere with imaging quality.
A spinal stabilization device can enhance imaging stability.

Purpose of Study

To achieve stable in vivo imaging of the mouse spinal cord.
To detail the dynamic behavior of cells in the spinal cord.
To facilitate time-lapse imaging of cellular interactions.

Methods Used

Anesthetizing the mouse with a mix for steady breathing.
Performing a laminectomy to expose the spinal cord.
Using a spinal stabilization device for imaging.
Isolating the exposed spinal cord segment for observation.

Main Results

Successful stabilization of the spinal column for imaging.
High-quality imaging data without the need for post-processing.
Detailed observations of cellular dynamics in the spinal cord.
Enhanced understanding of cell interactions in vivo.

Conclusions

The protocol allows for effective in vivo imaging of the spinal cord.
Minimizing respiratory movements is key to obtaining clear images.
This method can advance research on spinal cord dynamics.

Frequently Asked Questions

What is the main goal of this imaging protocol?

The main goal is to achieve stable in vivo imaging of the mouse spinal cord using two-photon microscopy.

How does the anesthetic regimen contribute to the procedure?

The anesthetic mix helps maintain a steady and calm breathing rhythm, minimizing respiratory-induced movements.

What is a laminectomy?

A laminectomy is a surgical procedure that involves removing a portion of the vertebra to expose the spinal cord for imaging.

Why is spinal stabilization important?

Spinal stabilization is crucial for reducing motion artifacts during imaging, leading to clearer and more reliable data.

What can researchers learn from this imaging technique?

Researchers can observe the dynamic behavior of cells and their interactions within the spinal cord over time.

Is post-processing required for the imaging data?

No, the method produces raw imaging data that require no alignment or other post-processing.

Un protocole minimalement invasive pour stabiliser la colonne vertébrale de souris et d'effectuer répétitives

Le but de cette procédure est d’effectuer une imagerie in vivo stable dans la moelle épinière de la souris à l’aide de la microscopie à deux photons. Ceci est accompli en anesthésant d’abord l’animal avec un mélange d’anesthésiques qui permet d’obtenir un rythme respiratoire stable mais calme. La deuxième étape de la procédure consiste à exposer la colonne vertébrale au niveau souhaité, puis à effectuer une laminectomie pour exposer la moelle épinière sous-jacente afin qu’elle puisse être imagée in vivo.

La troisième étape consiste à placer l’animal sur le dispositif de stabilisation de la colonne vertébrale. La dernière étape est l’isolement du segment exposé de la moelle épinière. En préparation de l’imagerie in vivo, on peut finalement obtenir des résultats qui détaillent le comportement dynamique des cellules et leurs interactions avec d’autres types de cellules ou structures de la moelle épinière vivante grâce à l’imagerie in vivo en accéléré à l’aide de la microscopie à deux photons.

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