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. 2018 Feb 7;8:2541. doi: 10.1038/s41598-018-20899-9

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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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Characterization of microcrystals. (A) HA crystals detected by light microscopy, UV-TPEF, and SHG (left to right). Scale bars: 50 μm. (B) Lysozyme microcrystals detected by UV-TPEF and light microscopy in the crystallization buffer, 0.1 M sodium acetate (pH 4.8), 18% NaCl, and 6% PEG 400. Supersaturation-controlled microcrystallization was examined by using evaporation times of 0 to 22 min in 2 min delay steps.