From Ultraviolet to Near-Infrared: Label-Free Reflection-Mode Hyperspectral Photoacoustic Microscopy for Single-Cell Biochemical Mapping
From Ultraviolet to Near-Infrared: Label-Free Reflection-Mode Hyperspectral Photoacoustic Microscopy for Single-Cell Biochemical Mapping
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DOI：
10.1089/genbio.2023.0035
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发表时间：
2023-10
期刊：
影响因子：
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通讯作者：
Qiangzhou Rong;Carlos Taboada;Ángela del Águila;Ilaria Merutka;N. Jayasundara;Yushun Zeng;Wei Yang;Qifa Zhou;Junjie Yao
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文献类型：
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作者：
Qiangzhou Rong;Carlos Taboada;Ángela del Águila;Ilaria Merutka;N. Jayasundara;Yushun Zeng;Wei Yang;Qifa Zhou;Junjie Yao
高光谱成像已成为一种有价值的技术，用于分析生物组织组成的探测内在或外源的生物分子。然而，传统的高光谱成像方法主要依赖于荧光特征，限制了它们在非荧光样品中的应用。为了克服这一限制，开发了一种无标记反射模式高光谱光声显微镜（RHS-PAM）系统。RHS-PAM能够使用从紫外到近红外的激发波长对具有宽范围内禀对比度的厚生物样品进行成像。RHS-PAM消除了对组织染色的需要，并在所有波长下实现了细胞级空间分辨率和自动图像共配准。RHS-PAM的概念验证应用已经在多种模式生物上得到证实，包括秀丽隐杆线虫、青蛙蝌蚪、斑马鱼和老鼠。该技术利用核酸、蛋白质、血红蛋白、黑色素和脂质的光学吸收对比，成功地对这些生物的丰富结构和分子特征进行了成像。结果表明，RHS-PAM能够提供丰富的光学对比度、高空间分辨率和扩展的无标签成像光谱范围。我们相信RHS-PAM是一种非常有前途的工具，可以用于多种生物组织的单细胞生化制图。
Hyperspectral imaging has emerged as a valuable technique for analyzing biological tissue compositions by probing intrinsic or exogenous biomolecules. However, conventional hyperspectral imaging methods predominantly rely on fluorescent signatures, limiting their application to nonfluorescent samples. To overcome this limitation, a label-free reflection-mode hyperspectral photoacoustic microscopy (RHS-PAM) system has been developed. RHS-PAM enables the imaging of thick biological samples with a wide range of intrinsic contrasts using excitation wavelengths ranging from ultraviolet to near infrared. RHS-PAM eliminates the need for tissue staining, and has achieved cellular-level spatial resolution and automatic image coregistrations at all wavelengths. Proof-of-concept applications of RHS-PAM have been demonstrated on various model organisms, includingCaenorhabditis elegans, frog tadpole, zebrafish, and mouse. The technique has successfully imaged a wealth of structural and molecular features in these organisms, utilizing the optical absorption contrast of nucleic acids, proteins, hemoglobin, melanin, and lipids. The results highlight the capability of RHS-PAM to provide rich optical contrast, high spatial resolution, and an extended spectral range for label-free imaging. We believe that RHS-PAM represents a highly promising tool for single-cell biochemical mapping of diverse biological tissues.