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Journal of Biomedical Optics

In situ visualization of intracellular morphology of epidermal cells using stimulated Raman scattering microscopy
Author(s): Mariko Egawa; Kyoya Tokunaga; Junichi Hosoi; Shinya Iwanaga; Yasuyuki Ozeki
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Paper 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.

Paper Details

Date Published: 31 August 2016
PDF: 10 pages
J. Biomed. Opt. 21(8) 086017 doi: 10.1117/1.JBO.21.8.086017
Published in: Journal of Biomedical Optics Volume 21, Issue 8
Show Author Affiliations
Mariko Egawa, Shiseido Co., Ltd. (Japan)
Kyoya Tokunaga, The Univ. of Tokyo (Japan)
Junichi Hosoi, Shiseido Co., Ltd. (Japan)
Shinya Iwanaga, Shiseido Co., Ltd. (Japan)
Yasuyuki Ozeki, The Univ. of Tokyo (Japan)

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