Overview

This protocol describes a detailed step-by-step approach for creating a chronic cranial window and microvascular embolism mouse model, optimized for in vivo two-photon microscopy imaging. The method utilizes fluorescent polystyrene microspheres to investigate the effects of capillary plugging.

Key Study Components

Area of Science

Neuroscience
Microvascular research
In vivo imaging techniques

Background

Understanding the consequences of microvascular occlusions is crucial for developing new treatment strategies.
Real-time imaging allows for tracking of fluorescent particles in the cerebral capillary network.
Previous studies have shown that capillary plugging can lead to local tissue damage.
This research aims to elucidate the mechanisms of embolus clearance in the brain.

Purpose of Study

To investigate the short and long-term effects of capillary plugging.
To understand how microvascular occlusions affect blood flow and the blood-brain barrier.
To explore potential treatment strategies for conditions related to microvascular damage.

Methods Used

Creation of a chronic cranial window in mice for imaging.
Injection of fluorescent microspheres to induce microvascular embolism.
Utilization of two-photon microscopy for real-time imaging.
Post-surgery imaging to confirm microsphere lodging in the brain.

Main Results

Microspheres were predominantly lodged in the ipsilateral hemisphere.
Increased microsphere counts were observed with the addition of Tween20.
Mice injected with Tween20 showed faster recovery post-surgery.
Fluorescence microscopy revealed occlusion of capillaries by microspheres.

Conclusions

This method allows for detailed study of microvascular occlusions in vivo.
Findings may lead to new insights into treatment strategies for cerebral microvascular conditions.
The study enhances understanding of the dynamics of blood flow and tissue damage in the brain.

Frequently Asked Questions

What is the significance of using fluorescent microspheres?

Fluorescent microspheres allow for real-time tracking of microvascular occlusions and their effects on cerebral blood flow.

How does this method contribute to neuroscience research?

It provides insights into the mechanisms of microvascular occlusions and potential therapeutic approaches for related conditions.

What imaging technique is employed in this study?

Two-photon microscopy is used for high-resolution in vivo imaging of the cerebral capillary network.

What are the expected outcomes of this research?

The research aims to clarify the effects of capillary plugging on local tissue damage and blood flow dynamics.

How long can the effects of microsphere injection be monitored?

The local consequences of microsphere injection can be followed for weeks after the procedure.

What role does Tween20 play in the microsphere mixture?

Tween20 enhances the effectiveness of the microsphere mixture, leading to better recovery and increased microsphere counts in the brain.

Institution - Amsterdam Neuroscience, neurovascular disorders

Articles Published in JoVE