Giant vesicles formed by gentle hydration and electroformation: A comparison by fluorescence microscopy
Giant vesicles formed by gentle hydration and electroformation: A comparison by fluorescence microscopy
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DOI：
10.1016/j.colsurfb.2005.01.010
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发表时间：
2005-05-10
影响因子：
5.8
通讯作者：
Cribier, S
中科院分区：
文献类型：
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作者：
Rodriguez, N;Pincet, F;Cribier, S
巨大的单层囊泡（直径为几十微米）通常由干燥脂质膜的水合作用产生。加入水溶液后，使用两种主要方案：（i）温和水合法，其中囊泡自发形成;和（ii）电铸法，其中施加交流电场。已知电形成可提高囊泡的单层率，尽管它对囊泡的微生物组成施加了更多限制条件。在这里，我们通过使用荧光显微镜进一步表征这些方法。它不仅能够灵敏地检测缺陷，而且能够评估这些缺陷中的脂质量。提出了一个分类的缺陷和统计数据的相对重要性方面的方法和脂质组合物：它显示，例如，80%的囊泡从98%的1，2-二油酰基-sn-甘油-3-磷酸胆碱通过电铸获得的没有对只有40%的囊泡与温和的水合方法的显着缺陷。它还表明，存在太多的带负电荷的脂质不利于形成单层囊泡与这两种方法。对于温和的水合，我们检查了带负电荷的脂质是否以与形成它们的脂质混合物相同的比例插入囊泡膜中。尽管带负电荷的1，2-二油酰基-sn-甘油-3-[磷酸-L-丝氨酸]的存在增加，但带负电荷的标记脂质的恒定掺入倾向于证实囊泡的组成确实接近于初始混合物的组成。(c)2005年由Elsevier B. V.出版
Giant unilamellar vesicles (diameter of a few tens of micrometers) are commonly produced by hydration of a dried lipidic film. After addition of the aqueous solution, two major protocols are used: (i) the gentle hydration method where the vesicles spontaneously form and (ii) the electroformation method where an ac electric field is applied. Electroformation is known to improve the rate of unilamellarity of the vesicles though it imposes more restricting conditions for the lipidic composition of the vesicles. Here we further characterize these methods by using fluorescence microscopy. It enables not only a sensitive detection of the defects but also an evaluation of the quantity of lipids in these defects. A classification of the defects is proposed and statistics of their relative importance in regard to both methods and lipid composition are presented: it shows for example that 80% of the vesicles obtained by electroformation from 98% 1,2-Dioleoyl-sn-Glycero-3-Phosphocholine are devoid of significant defects against only 40% of the vesicles with the gentle hydration method. It is also shown that the presence of too many negatively charged lipids does not favor the formation of unilamellar vesicles with both methods. For the gentle hydration, we checked if the negatively charged lipids were inserted in the vesicles membrane in the same proportion as that of the lipid mixture from which they are formed. The constant incorporation of a negatively charged labeled lipid despite an increasing presence of negatively charged 1,2-Dioleoyl-sn-Glycero-3-[Phospho-L-Serine] tends to confirm that the composition of vesicles is indeed close to that of the initial mixture. (c) 2005 Published by Elsevier B.V.