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. 2021 Mar 4;22(5):2574. doi: 10.3390/ijms22052574

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Atomic force microscopy (AFM) height images of supported lipid bilayers. Images of phase separation in binary and ternary lipid bilayers of DOPC:SM:Chol (1,2-dioleoyl-sn-glycero-3-phosphocholine:sphingomyelin:cholesterol) or 25HC (top), and respective height differences between the two membrane phases and their area percentage of the whole supported lipid bilayer (SLB), calculated by the fit of a sum of two Gaussians (bottom). (A) Bilayer with 20% Chol (DOPC:SM:Chol 2:2:1 in molar ratio, corresponding to 4.32:4.10:1 in molar volume ratios) exhibiting phase separation of liquid-ordered (L_o) smaller domains. (B) Larger-sized L_o domains in a phase separation with Chol content increased to 33% (DOPC:SM:Chol 1:1:1 in molar ratio; 2.16:2.05:1 in molar volume ratios). (C) Bilayers with 33% 25HC (DOPC:SM:25HC 1:1:1 in molar ratio; 2.16:2.05:1 in molar volume ratios), displaying smaller phase separated domains. (D) Bilayer with 50% SM (DOPC:SM 1:1 in molar ratio; 1.05:1 in molar volume ratios) exhibiting a phase separation of solid-ordered (s_o) domains. All images are 30 × 30 µm².