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. 2019 Apr 3;9:5561. doi: 10.1038/s41598-019-41466-w

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© The Author(s) 2019

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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Human fetal femur-derived skeletal cells were cultured as a three-dimensional pellet over 21 days in chondrogenic media to generate cartilage tissue. (a) Raman spectrum in the ‘fingerprint’ region. The three marked bands were targeted for CARS imaging. (b) CARS images acquired at the ‘fingerprint’ vibrations of: 1061 cm⁻¹ (ν_s(OSO₃⁻)), 1450 cm⁻¹ (δ(CH₂)), and 1668 cm⁻¹ (ν(C=O)). Scale bars correspond to 50 µm.