Polarized THG microscopy identifies compositionally different lipid droplets in mammalian cells.
Polarized THG microscopy identifies compositionally different lipid droplets in mammalian cells.
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DOI：
10.1016/j.bpj.2014.10.009
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发表时间：
2014-11-18
影响因子：
3.4
通讯作者：
Kauranen, Martti
中科院分区：
文献类型：
--
作者：
Bautista, Godofredo;Pfisterer, Simon G.;Huttunen, Mikko J.;Ranjan, Sanjeev;Kanerva, Kristiina;Ikonen, Elina;Kauranen, Martti
细胞将多余的脂质储存为两种主要化合物，三酰甘油（TAG）和胆固醇酯（CE），在脂滴（LD）内。脂质有序化的程度被认为在LD中脂质的流动性和酶促加工中起主要作用。在这里，我们提供的证据表明，偏振三次谐波产生（THG）显微镜区分天然TAG和CE富集LD在细胞中，由于两种脂质的不同顺序。我们首先证明，从合成TAG和CE富集LD使用三倍频显微镜与线性和圆偏振的响应不同，根据其不同的内在顺序。然后，我们采用模拟解剖偏振效应如何影响三倍频从各向同性LD。最后，我们在小鼠巨噬细胞中诱导TAG和CE富集的LD，并证明极化THG反应随着CE/TAG比率的增加以非线性方式增加。这表明，随着CE含量的增加，有一个相当尖锐的过渡增加LD订购。我们的研究结果表明，偏振三倍频显微镜能够无标记的定量分析LD排序和区分组成不同的LD在完整的哺乳动物细胞。
Cells store excess lipids as two major compounds, triacylglycerols (TAGs) and cholesteryl esters (CEs), inside lipid droplets (LDs). The degree of lipid ordering is considered to play a major role in the mobility and enzymatic processing of lipids in LDs. Here, we provide evidence that polarized third-harmonic generation (THG) microscopy distinguishes between native TAG- and CE-enriched LDs in cells due to the different ordering of the two lipid species. We first demonstrate that the responses from synthetic TAG- and CE-enriched LDs using THG microscopy with linear and circular polarizations differ according to their different intrinsic ordering. We then employ simulations to dissect how polarization effects influence the THG from an isotropic LD. Finally, we induce TAG- and CE-enriched LDs in murine macrophages and demonstrate that polarized THG responses increase in a nonlinear fashion with increasing CE/TAG ratio. This suggests that with an increasing CE content, there is a rather sharp transition toward increased LD ordering. Our results demonstrate that polarized THG microscopy enables label-free quantitative analysis of LD ordering and discriminates between compositionally different LDs in intact mammalian cells.
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影响因子：
64.5
作者：
Moore KJ;Tabas I
通讯作者：
Tabas I
影响因子：
3.6
作者：
Débarre, D;Supatto, W;Beaurepaire, E
通讯作者：
Beaurepaire, E
影响因子：
3.4
作者：
Debarre, Delphine;Beaurepaire, Emmanuel
通讯作者：
Beaurepaire, Emmanuel
影响因子：
1.9
作者：
Bowden JA;Shao F;Albert CJ;Lally JW;Brown RJ;Procknow JD;Stephenson AH;Ford DA
通讯作者：
Ford DA
影响因子：
8.6
作者：
Verbiest, T;Kauranen, M;Persoons, A
通讯作者：
Persoons, A