Real-Time Intravital Multiphoton Microscopy to Visualize Focused Ultrasound and Microbubble Treatments to Increase Blood-Brain Barrier Permeability

作者： Charissa Poon1,2, Melina Mühlenpfordt*3, Marieke Olsman*3, Spiros Kotopoulis4,5, Catharina de Lange Davies3, Kullervo Hynynen1,2,6 1Physical Sciences Platform,Sunnybrook Research Institute, 2Institute of Biomedical Engineering,University of Toronto, 3Department of Physics,Norwegian University of Science and Technology, 4Department of Clinical Medicine,University of Bergen, 5Exact Therapeutics AS, 6Department of Medical Biophysics,University of Toronto

简介：

Overview

This protocol outlines the procedures for real-time in vivo multiphoton fluorescence imaging of the rodent brain during focused ultrasound and microbubble treatments. This technique aims to enhance blood-brain barrier permeability, facilitating drug delivery for brain diseases.

Key Study Components

Area of Science

• Neuroscience
• Biomedical Engineering
• Imaging Techniques

Background

• Ultrasound and microbubbles can transiently increase blood-brain barrier permeability.
• This method allows for better drug delivery to the brain.
• Traditional imaging methods do not capture real-time responses during treatment.
• Intravital multiphoton imaging provides a solution for real-time observation.

Purpose of Study

• To demonstrate a protocol for concurrent ultrasound treatment and multiphoton imaging.
• To enable real-time monitoring of blood-brain barrier permeability changes.
• To evaluate the effectiveness of drug delivery methods in preclinical models.

Methods Used

• Preparation of cranial window in mice for imaging.
• Use of a ring transducer for ultrasound application.
• Injection of microbubbles and fluorescent dextran for imaging.
• Real-time imaging of vascular changes during treatment.

Main Results

• Successful imaging of blood-brain barrier permeability changes.
• Quantitative analysis of dextran leakage as a drug delivery model.
• Real-time monitoring of vascular responses to ultrasound treatment.
• High spatial and temporal resolution achieved in imaging.

Conclusions

• The protocol allows for effective real-time imaging of the rodent brain.
• Ultrasound microbubble treatment can enhance drug delivery across the blood-brain barrier.
• This method can be applied to various research models and ultrasound applications.

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