超高信息含量化学成像与宽带连贯抗斯托克斯拉曼和两光子光寿命显微镜
Haoyu Xu1, Wei-Wen Chen2, Jessica Z Dixon3
1Department of Biomedical Engineering, Georgia Institute of Technology.
Journal of visualized experiments : JoVE
|October 27, 2025
概括
这项研究引入了一个多式成像平台,将拉曼光谱和光终身成像结合起来,用于体内代谢分析. 该系统通过整合化学和生命周期数据来提高亚细胞分析的准确性,改善生物解释.
科学领域:
- 生物光子学 生物光子学
- 频谱学是一种光谱学.
- 显微镜的使用方法
背景情况:
- 拉曼指纹光谱和光终身成像 (FLIM) 是分析生物代谢资料的先进技术.
- 这些方法分别探测分子振动和光体兴奋状态寿命,为细胞化学环境提供了洞察力.
研究的目的:
- 展示一个新的多式成像平台,集成宽带连贯抗斯托克斯拉曼散射 (BCARS) 和两光子光终身成像 (2p-FLIM).
- 为了证明在体内同时获得拉曼和FLIM信号以进行亚细胞代谢表征.
- 通过结合化学和光寿命数据,提高生物系统生物解释的准确性.
主要方法:
- 开发一个结合BCARS和2p-FLIM的多模式显微镜平台.
- 同时在活体中从表达绿色光蛋白 (GFP) 的活体Caenorhabditis elegans (C. elegans) 获得数据.
- 交叉模式成像分析比较BCARS,2p-FLIM和两光子激发光 (TPEF) 信号.
主要成果:
- 成功同时获得生物相关的拉曼指纹光谱和光生命周期信号 in vivo.
- 展示BCARS和2p-FLIM在表征亚细胞区和区分它们之间的能力.
- 通过比较活体C. elegans中染色器官的信号来验证多模式方法的有效性,仅通过光成像来验证潜在的错误阳性.
结论:
- 开发的BCARS/2p-FLIM平台可以在体内以亚细胞分辨率同时进行化学和光终身成像.
- 这种跨模式成像方法显著提高了复杂的生物系统中的生物解释的准确性.
- 该协议为先进的代谢分析和生物标本分析提供了一个强大的框架.
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