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. 2018 Mar 14;8:4476. doi: 10.1038/s41598-018-22264-2

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Imaging of fresh (not-frozen), normal human skin discarded during a Mohs surgical procedure. (a) Air immersion MUSE, (b) water immersion MUSE, (c) NLM and (d) FFPE H&E. The epidermis and multiple hair follicles can be seen using each modality. Magnified images of the epidermis layer (e) air immersion MUSE, (f) water immersion MUSE, (g) NLM, (h) FFPE H&E demonstrate that both air and water immersion MUSE reproduce the appearance of the normal epidermis layer, while the thinner optical sectioning of water immersion MUSE enables visualization of finer details than air immersion. Magnified images of a hair follicle and glands (i) air immersion MUSE, (j) water immersion MUSE, (k) NLM, (l) FFPE H&E demonstrate that both air and water immersion MUSE enable visualization of structures of the hair follicle (hair, sheath and glands), although some glandular structures are less clear in MUSE images because of the stacked appearance of surface and subsurface cell layers. Scale bar 1 mm (a–d), 100 µm (e–h), 200 µm (i–l). Diffuse EY can be seen both in the MUSE images and in the NLM image. However, the effect is more pronounced in the MUSE image because of the thick optical sectioning. Exposure time of MUSE: 0.3 s (water), 0.7 s (air). Air immersion MUSE: https://goo.gl/LCXYKT. Water immersion MUSE: https://goo.gl/TGfSCP. NLM: https://goo.gl/3Z66ED. FFPE H&E: https://goo.gl/UkuVfA.