利用生物发光显微镜研究活体小鼠大脑中氧动态的方案。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2024-12-20 Epub Date: 2024-09-26 DOI:10.1016/j.xpro.2024.103334

Antonios Asiminas, Ryszard S Gomolka, Stefanie Gregoriades, Hajime Hirase, Maiken Nedergaard, Felix R M Beinlich

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引用次数: 0

摘要

生物发光成像（BLI）依赖于底物和酶之间的生化反应，这种反应在会聚时引发光发射。在这里，我们介绍一种利用荧光素酶与其底物之间的氧依赖性反应研究体内小鼠大脑中分子氧动态的方案。我们描述了急性开颅手术、病毒转染、底物给药、成像和缺氧袋分析的步骤。与现有的电极和磷光等方法相比，该方案具有更优越的时空特性。有关本方案使用和执行的完整细节，请参阅 Beinlich 等人的文章1。

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Protocol to study oxygen dynamics in the in vivo mouse brain using bioluminescence microscopy.

Bioluminescence imaging (BLI) relies on the biochemical reaction between substrate and enzyme that triggers light emission upon convergence. Here, we present a protocol to study molecular oxygen dynamics in the in vivo mouse brain using the oxygen-dependent reaction between luciferase and its substrate. We describe steps for acute craniotomy, viral transfection, substrate administration, imaging, and analysis of hypoxic pockets. This protocol offers superior spatiotemporal properties compared to established approaches like electrodes and phosphorescence. For complete details on the use and execution of this protocol, please refer to Beinlich et al.¹.

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