Research Papers: Imaging

In situ visualization of intracellular morphology of epidermal cells using stimulated Raman scattering microscopy

[+] Author Affiliations
Mariko Egawa, Junichi Hosoi, Shinya Iwanaga

Shiseido Global Innovation Center, 2-2-1 Hayabuchi, Tsuzuki-ku, Yokohama, Kanagawa 224-0025, Japan

Kyoya Tokunaga, Yasuyuki Ozeki

The University of Tokyo, Department of Electrical Engineering and Information Systems, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan

J. Biomed. Opt. 21(8), 086017 (Aug 31, 2016). doi:10.1117/1.JBO.21.8.086017
History: Received April 12, 2016; Accepted August 10, 2016
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Abstract.  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.

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© 2016 Society of Photo-Optical Instrumentation Engineers

Citation

Mariko Egawa ; Kyoya Tokunaga ; Junichi Hosoi ; Shinya Iwanaga and Yasuyuki Ozeki
"In situ visualization of intracellular morphology of epidermal cells using stimulated Raman scattering microscopy", J. Biomed. Opt. 21(8), 086017 (Aug 31, 2016). ; http://dx.doi.org/10.1117/1.JBO.21.8.086017


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