Protocol to assess extravasation of fluorescent molecules in mice after ultrasound-mediated blood-brain barrier opening

Lea Peko; Sharon Katz; Roni Gattegno; Tali Ilovitsh

doi:10.1016/j.xpro.2023.102770

Protocol to assess extravasation of fluorescent molecules in mice after ultrasound-mediated blood-brain barrier opening

Lea Peko, Sharon Katz, Roni Gattegno, Tali Ilovitsh^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

Blood-brain barrier disruption (BBBD) using focused ultrasound (FUS) and microbubbles (MBs) is an effective tool for therapeutic delivery to the brain. Here, we present an optimized protocol for quantifying fluorescent molecules extravasation in mice. We describe steps for ultrasound treatment, injection of MBs and fluorescent dyes, brain harvesting, microscopy imaging, and image postprocessing algorithm. Our protocol has proven to successfully conduct a diameter-dependent analysis that measures vascular leakage following FUS-mediated BBBD at a single blood vessel resolution. For complete details on the use and execution of this protocol, please refer to Katz et al.¹

Original language	English
Article number	102770
Journal	STAR Protocols
Volume	5
Issue number	1
DOIs	https://doi.org/10.1016/j.xpro.2023.102770
State	Published - 15 Mar 2024

Keywords

Biotechnology and bioengineering
Microscopy
Neuroscience
Physics

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10.1016/j.xpro.2023.102770

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abstract = "Blood-brain barrier disruption (BBBD) using focused ultrasound (FUS) and microbubbles (MBs) is an effective tool for therapeutic delivery to the brain. Here, we present an optimized protocol for quantifying fluorescent molecules extravasation in mice. We describe steps for ultrasound treatment, injection of MBs and fluorescent dyes, brain harvesting, microscopy imaging, and image postprocessing algorithm. Our protocol has proven to successfully conduct a diameter-dependent analysis that measures vascular leakage following FUS-mediated BBBD at a single blood vessel resolution. For complete details on the use and execution of this protocol, please refer to Katz et al.1",

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AB - Blood-brain barrier disruption (BBBD) using focused ultrasound (FUS) and microbubbles (MBs) is an effective tool for therapeutic delivery to the brain. Here, we present an optimized protocol for quantifying fluorescent molecules extravasation in mice. We describe steps for ultrasound treatment, injection of MBs and fluorescent dyes, brain harvesting, microscopy imaging, and image postprocessing algorithm. Our protocol has proven to successfully conduct a diameter-dependent analysis that measures vascular leakage following FUS-mediated BBBD at a single blood vessel resolution. For complete details on the use and execution of this protocol, please refer to Katz et al.1

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Protocol to assess extravasation of fluorescent molecules in mice after ultrasound-mediated blood-brain barrier opening

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