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

Lea Peko, Sharon Katz, Roni Gattegno, Tali Ilovitsh

Research output: Contribution to journalArticlepeer-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 languageEnglish
Article number102770
JournalSTAR Protocols
Volume5
Issue number1
DOIs
StatePublished - 15 Mar 2024
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2023 The Author(s)

Funding

This work was supported by funding from Insightec Ltd. the Israel Science Foundation (grant numbers 192/ 22 and 3450/20), the Israel Ministry of Science and Technology (grant number 101716), an ERC StG grant (no. 101041118) (NanoBubbleBrain), and a Zimin Institute grant, and was partially supported by a grant from the Nicholas and Elizabeth Slezak Super Center for Cardiac Research and Biomedical Engineering at Tel Aviv University. L.P. assisted with the in vivo experiments, microscopy, and image processing and wrote this manuscript. S.K. designed and performed the research. R.G. revised the manuscript. T.I. advised and designed the research. All authors have approved the final version of this manuscript. The authors declare no competing interests. This work was supported by funding from Insightec Ltd., the Israel Science Foundation (grant numbers 192/ 22 and 3450/20 ), the Israel Ministry of Science and Technology (grant number 101716 ), an ERC StG grant (no. 101041118 ) (NanoBubbleBrain), and a Zimin Institute grant, and was partially supported by a grant from the Nicholas and Elizabeth Slezak Super Center for Cardiac Research and Biomedical Engineering at Tel Aviv University .

FundersFunder number
Insightec Ltd.
Insightec Ltd. the Israel Science Foundation
European Commission101041118
Israel Science Foundation192/ 22, 3450/20
Tel Aviv University
Ministry of science and technology, Israel101716
Nicholas and Elizabeth Slezak Super Center for Cardiac Research and Biomedical Engineering

    Keywords

    • Biotechnology and bioengineering
    • Microscopy
    • Neuroscience
    • Physics

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