In situ visualization of intracellular morphology of epidermal cells using stimulated Raman scattering microscopy
In situ visualization of intracellular morphology of epidermal cells using stimulated Raman scattering microscopy
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DOI：
10.1117/1.jbo.21.8.086017
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发表时间：
2016-08-01
影响因子：
3.5
通讯作者：
Ozeki, Yasuyuki
中科院分区：
文献类型：
--
作者：
Egawa, Mariko;Tokunaga, Kyoya;Ozeki, Yasuyuki
表皮细胞的可视化是重要的，因为角质形成细胞（KCs）的分化模式被认为与皮肤的功能和状态有关。光学显微镜已被广泛用于研究表皮细胞，但其适用性仍然是有限的，因为需要样品固定和染色。在这里，我们报告了我们的组织和文化中的表皮细胞的无染色观察受激拉曼散射（SRS）显微镜，提供分子振动对比。SRS使我们能够观察到皮肤组织中的各种细胞形态，包括棘层中的梯状结构、颗粒层中的KC的去核以及角质层中的三维细胞柱结构。我们注意到棘层中的一些细胞在细胞质中具有比KCs更亮的信号。为了检验表皮层观察的相关性，我们还观察了培养的表皮细胞，包括处于不同分化阶段的KC、黑素细胞和朗格汉斯细胞样细胞。他们的SRS图像还显示了各种形态，表明在组织中观察到的形态差异与细胞谱系相对应。这些结果表明，可能应用SRS显微镜皮肤病学调查的细胞谱系和类型的表皮细胞水平的分析。(C)2016年，美国光电仪器工程师学会（SPIE）
Visualization of epidermal cells is important because the differentiation patterns of keratinocytes (KCs) are considered to be related to the functions and condition of skin. Optical microscopy has been widely used to investigate epidermal cells, but its applicability is still limited because of the need for sample fixation and staining. Here, we report our staining-free observation of epidermal cells in both tissue and culture by stimulated Raman scattering (SRS) microscopy that provides molecular vibrational contrast. SRS allowed us to observe a variety of cellular morphologies in skin tissue, including ladder-like structures in the spinous layer, enucleation of KCs in the granular layer, and three-dimensional cell column structures in the stratum corneum. We noticed that some cells in the spinous layer had a brighter signal in the cytoplasm than KCs. To examine the relevance of the observation of epidermal layers, we also observed cultured epidermal cells, including KCs at various differentiation stages, melanocytes, and Langerhans cell-like cells. Their SRS images also demonstrated various morphologies, suggesting that the morphological differences observed in tissue corresponded to the cell lineage. These results indicate the possible application of SRS microscopy to dermatological investigation of cell lineages and types in the epidermis by cellular-level analysis. (C) 2016 Society of Photo-Optical Instrumentation Engineers (SPIE)