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Isolation and Imaging of Microvessels From Brain Tissue
Josephine K Buff1,2, Carolyn R Bertozzi3,4,5, Tony Wyss-Coray1,2,4,6
1Department of Neurology and Neurological Sciences, Stanford University School of Medicine, Stanford, CA, USA.
Bio-Protocol
|September 15, 2025
Summary
This study presents a new protocol for isolating brain microvessels, crucial for understanding the blood-brain barrier (BBB). This method allows for detailed molecular analysis of BBB integrity in various conditions.
Area of Science:
- Neuroscience
- Vascular Biology
- Biochemistry
Background:
- The blood-brain barrier (BBB) is vital for brain function, comprising specialized microvessels.
- Studying BBB composition and function requires reliable isolation of these microvessels.
- Current methods may limit comparative analyses across species or disease states.
Purpose of the Study:
- To develop and validate a protocol for isolating intact microvessels from human and murine brain tissue.
- To enable molecular and imaging analyses of microvessels for studying BBB integrity.
- To facilitate translational research by allowing direct comparisons across species and disease models.
Main Methods:
- Mechanical dissociation of brain tissue.
- Density-based separation to isolate microvessels.
- Application to fresh or frozen human and murine brain samples.
Main Results:
- Isolated microvessels maintain molecular integrity.
- The protocol is compatible with downstream applications like fluorescence imaging, RNA sequencing, proteomics, western blotting, and ELISA.
- The method was successfully applied to study molecular changes in neurodegeneration and aging.
Conclusions:
- This protocol provides a robust method for microvessel isolation from diverse brain tissues.
- The isolated microvessels are suitable for comprehensive molecular and imaging analyses.
- This technique supports translational research by enabling cross-species and cross-disease comparisons of BBB biology.
