Time-Resolved Mid-Infrared Photothermal Microscopy for Imaging Water-Embedded Axon Bundles.
Time-Resolved Mid-Infrared Photothermal Microscopy for Imaging Water-Embedded Axon Bundles.
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DOI：
10.1021/acs.analchem.3c02352
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发表时间：
2023-11-14
影响因子：
7.4
通讯作者：
Sander, Michelle Y.
中科院分区：
文献类型：
--
作者：
Samolis, Panagis D.;Zhu, Xuedong;Sander, Michelle Y.
由于水的高红外吸收，与主要的蛋白质和脂质带重叠，在富含水的环境中存在很少的实验工具用于执行生物样品的无标记成像。介绍了一种基于时间分辨中红外光热显微镜的新型成像模式，并将其应用于盐水浴环境中的轴突束成像。光热诱导的空间梯度的轴突束膜与盐水和周围的生物组织的接口进行观察，并通过高速boxcar检测系统的时间特征。本地化的时间配置文件与增强的信噪比，超时间图像堆栈，和二维映射的时间衰减配置文件的采集，而不需要复杂的后图像处理。轴突束被发现有一个更大的时间衰减分布相比，水的背景，允许这些瞬态动力学的基础上的背景分化。随着时间的推移的信号演变的定量分析允许表征在不同区域的热约束的水平。当轴突束被复杂的异质组织包围时，其包含较小的特征，与水环境相比，观察到更强的热约束，从而揭示了水生物界面的热传递动力学。
Few experimental tools exist for performing label-free imaging of biological samples in a water-rich environment due to the high infrared absorption of water, overlapping with major protein and lipid bands. A novel imaging modality based on time-resolved mid-infrared photothermal microscopy is introduced and applied to imaging axon bundles in a saline bath environment. Photothermally induced spatial gradients at the axon bundle membrane interfaces with saline and surrounding biological tissue are observed and temporally characterized by a high-speed boxcar detection system. Localized time profiles with an enhanced signal-to-noise, hyper-temporal image stacks, and two-dimensional mapping of the time decay profiles are acquired without the need for complex post image processing. Axon bundles are found to have a larger distribution of time decay profiles compared to the water background, allowing background differentiation based on these transient dynamics. The quantitative analysis of the signal evolution over time allows characterizing the level of thermal confinement at different regions. When axon bundles are surrounded by complex heterogeneous tissue, which contains smaller features, a stronger thermal confinement is observed compared to a water environment, thus shedding light on the heat transfer dynamics across aqueous biological interfaces.
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