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. 2018 Oct 8;8:14974. doi: 10.1038/s41598-018-33226-z

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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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Mechanical properties of healthy RBCs at PL values for DMSO, glycerol and sodium azide: Healthy RBCs were treated at PL, 10*PL and 20*PL values of (A) Glycerol, (B) DMSO and (C) Sodium azide and membrane shear modulus determined through micropipette aspiration technique. A total sample size of at least 20 cells per measurement was taken for individual experiments. The cell membrane was monitored by Olympus IX71 microscope and processed by QCapture Pro 6.0. The maximum time span before the whole sample was replaced by a fresh sample was 2 hours. From the high-resolution recordings, the leading edge of the aspired RBC membrane was tracked manually for calculating the elastic shear modulus using the Hochmuth model. All RBCs treated at PL value showed no significant changes in the membrane deformability and were observed to have comparable shear modulus to control population (untreated control). Significant results are indicated as follows: ****<0.0001; ***<0.001; **0.001 to 0.01; *0.01 to 0.05. All other comparisons show no significant differences (Kruskal-Wallis test with Dunn’s multiple comparison test).