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Cardiovascular Engineering

Cardiovascular Engineering - Poster Session E

S18 - A Protocol for Isolation and Imaging of Cerebral Microvascular Networks

Saturday, October 11, 2025
10:00 AM - 11:00 AM PDT
Location: Exhibit Hall F, G & H

Introduction: : Precise regulation of cerebral blood flow (CBF) is essential for brain health, with dysregulation linked to stroke and neurodegenerative diseases [3]. Progress in understanding microvascular mechanisms has been limited by challenges in isolating and characterizing intact cerebral microvascular networks [2]. Addressing these limitations is critical for developing advanced disease models and novel therapies.

Materials and

Methods: : We utilized and optimized an experimental protocol for isolating mouse brain microvascular vessels (REF Dabertrand) . The procedure entails mouse brain isolation and dissection of a segment of the pial arteriole tree stemming from the Middle Cerebral Artery.  Attached Penetrating Arterioles (PAs) and capillaries are carefully cleaned from parenchymal tissue with microforcepts. The isolated vascular network is stretched by placement on the top of the solution in the dissecting dish and is picked by a coverslip for transporting it into a perfusion chamber for fluorescence microscopy or electrical recordings

Results, Conclusions, and Discussions:: This approach allowed to reproducibly extract segments of the arteriole network with attached capillaries. The freshly isolated vascular network is ideally suited for investigating signaling between arterioles and capillaries including how electrical and Calcium signals propagate along the vasculature. Preliminary data demonstrate the feasibility of capturing membrane potential changes along the vascular network using fluorescence microscopy (Fluovolt, Molecular Probes [1]) or Sharp electrodes and future work will characterize responses and signal propagation following focal pharmaco-mechanical stimulation. The ex vivo prep will be utilized for comparing responses between healthy and diseased tissue. The proposed protocol provides an ex vivo model for studying cerebral vascular function in health and in cerebrovascular disease.

Acknowledgements and/or References (Optional): : Acknowledgement
The work is supported by the National Institute of Neurological Disorders and Stroke, and the
National Institute of Aging of the NIH under award number R01NS119971.
References
1. Thermo Fisher Scientific. (n.d.). FluoVolt™ Membrane Potential Kit. Retrieved October
21, 2024, from https://www.thermofisher.com/order/catalog/product/F10488
2. Rosehart, A. C. et al., (2019). Ex Vivo Pressurized Hippocampal Capillary-Parenchymal
Arteriole Preparation for Functional Study. Journal of visualized experiments: JoVE,
(154), 10.3791/60676. https://doi.org/10.3791/60676
3. Moshkforoush , A et al., (2020, June 19). The capillary Kir channel as sensor and amplifier
of neuronal signals: Modeling insights on K+-mediated neurovascular communication.”
PNAS. https://www.pnas.org/doi/10.1073/pnas.2000151117