Photothermal Microscopy for High Sensitivity and High Resolution Absorption Contrast Imaging of Biological Tissues
Photothermal Microscopy for High Sensitivity and High Resolution Absorption Contrast Imaging of Biological Tissues
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DOI：
10.3390/photonics4020032
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发表时间：
2017-04
期刊：
影响因子：
2.4
通讯作者：
J. Miyazaki;Takayoshi Kobayahsi
中科院分区：
文献类型：
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作者：
J. Miyazaki;Takayoshi Kobayahsi
光热显微镜可用于可视化生物样本中非荧光色素蛋白的分布。在这里，我们开发了一种高灵敏度和高分辨率的光热显微镜，以低成本和紧凑的激光二极管作为光源。提出了一种新的检测方案，可将信噪比提高 4 倍以上。事实证明，光热显微镜的空间分辨率几乎是传统宽场显微镜的两倍。此外，我们证明了通过同时多波长成像来区分或识别生物分子的能力。对小鼠脑组织进行同步光热和荧光成像，以可视化表达黄色荧光蛋白和内源性非荧光发色团的神经元。
Photothermal microscopy is useful to visualize the distribution of non-fluorescence chromoproteins in biological specimens. Here, we developed a high sensitivity and high resolution photothermal microscopy with low-cost and compact laser diodes as light sources. A new detection scheme for improving signal to noise ratio more than 4-fold is presented. It is demonstrated that spatial resolution in photothermal microscopy is up to nearly twice as high as that in the conventional widefield microscopy. Furthermore, we demonstrated the ability for distinguishing or identifying biological molecules with simultaneous muti-wavelength imaging. Simultaneous photothermal and fluorescence imaging of mouse brain tissue was conducted to visualize both neurons expressing yellow fluorescent protein and endogenous non-fluorescent chromophores.