Hyperspectral Oblique Plane Microscopy Enables Spontaneous, Label-Free Imaging of Biological Dynamic Processes in Live Animals
Hyperspectral Oblique Plane Microscopy Enables Spontaneous, Label-Free Imaging of Biological Dynamic Processes in Live Animals
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高光谱斜平面显微镜能够对活体动物的生物动态过程进行自发、无标记成像
DOI：
10.1101/2023.03.15.532804
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Guo K
中科院分区：
文献类型：
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作者：
Guo K
自发拉曼成像已成为研究药物和生物样本分子组成的强大无标记技术。尽管拉曼成像可以促进对体内复杂生物现象的理解，但当前的成像方式在速度和样品兼容性方面受到限制。在这里，我们介绍了一种单物镜线扫描光片显微镜，名为-OPM，它可以在分钟到秒的时间尺度上记录拉曼图像。为了演示其功能，我们使用-OPM 根据微塑料颗粒的拉曼光谱特征来绘制和识别微塑料颗粒。在活体斑马鱼胚胎中，我们证明-OPM 可以以五分钟的间隔捕获伤口动态，揭示受伤区域细胞和细胞外基质组成的快速变化。最后，我们使用-OPM 同步并平均 36,800 个单独的帧，以有效每秒 28 帧的速度获得斑马鱼胚胎跳动心脏的高光谱视频，记录整个心动周期的成分变化。
Spontaneous Raman imaging has emerged as powerful label-free technique for investigating the molecular composition of medicines and biological specimens. Although Raman imaging can facilitate understanding of complex biological phenomena in vivo, current imaging modalities are limited in speed and sample compatibility. Here, we introduce a single-objective line-scanning light-sheet microscope, named-OPM, which records Raman images on a timescale of minutes to seconds. To demonstrate its function, we use-OPM to map and identify microplastic particles based on their Raman spectral characteristics. In live zebrafish embryos, we show that-OPM can capture wound dynamics at five-minute intervals, revealing rapid changes in cellular and extracellular matrix composition in the wounded region. Finally, we use-OPM to synchronize and average 36,800 individual frames to obtain hyperspectral videos of a zebrafish embryo’s beating heart at an effective 28 frames per second, recording compositional changes throughout the cardiac cycle.