小鼠和人脑微血管分离和RNA荧光可视化的方案。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2025-01-07 DOI:10.1016/j.xpro.2024.103530

Olivia M Osborne, Oandy Naranjo, Silvia Torices, Sarah Schmidlin, Destiny Tiburcio, Minseon Park, Michal Toborek

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引用次数: 0

摘要

在这里，我们提出了一种从小鼠和人的新鲜或快速冷冻脑组织中分离微血管的方案，然后利用RNAscope杂交技术可视化RNA以定量mRNA。我们描述了样品制备和分离，固定和杂交的步骤。该方案是专门设计与RNAscope在原位杂交整合。虽然该方案是在小鼠模型中开发的，但它可以优化用于其他生物体，包括人类大脑样本。

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Protocol for the isolation of brain microvessels and visualization of RNA fluorescence in mice and humans.

Here, we present a protocol for isolating microvessels from fresh or snap-frozen brain tissue from mice and humans, followed by visualization of RNA utilizing RNAscope hybridization for quantification of mRNA. We describe the steps for sample preparation and isolation, fixation, and hybridization. This protocol was specifically designed to integrate with RNAscope in situ hybridization. Although the protocol was developed in a mouse model, it can be optimized for use in other organisms, including human brain samples.

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