Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN 47907, USA.
J Lipid Res. 2010 Mar;51(3):672-7. doi: 10.1194/jlr.D000638. Epub 2009 Sep 23.
The ubiquity of lipids in biological structures and functions suggests that lipid metabolisms are highly regulated. However, current invasive techniques for lipid studies prevent characterization of the dynamic interactions between various lipid metabolism pathways. Here, we describe a noninvasive approach to study lipid metabolisms using a multifunctional coherent anti-Stokes Raman scattering (CARS) microscope. Using living Caenorhabditis elegans as a model organism, we report label-free visualization of coexisting neutral and autofluorescent lipid species. We find that the relative expression level of neutral and autofluorescent lipid species can be used to assay the genotype-phenotype relationship of mutant C. elegans with deletions in the genes encoding lipid synthesis transcription factors, LDL receptors, transforming growth factor beta receptors, lipid desaturation enzymes, and antioxidant enzymes. Furthermore, by coupling CARS with fingerprint confocal Raman analysis, we analyze the unsaturation level of lipids in wild-type and mutant C. elegans. Our study shows that complex genotype-phenotype relationships between lipid storage, peroxidation, and desaturation can be rapidly and quantitatively analyzed in a single living C. elegans.
脂质在生物结构和功能中的普遍性表明脂质代谢受到高度调控。然而，目前用于脂质研究的侵入性技术阻碍了对各种脂质代谢途径之间动态相互作用的描述。在这里，我们描述了一种使用多功能相干反斯托克斯拉曼散射（CARS）显微镜研究脂质代谢的非侵入性方法。我们使用活体秀丽隐杆线虫作为模型生物，报告了非标记可视化共存的中性和自发荧光脂质种类。我们发现，中性和自发荧光脂质种类的相对表达水平可用于测定缺失编码脂质合成转录因子、低密度脂蛋白受体、转化生长因子β受体、脂质去饱和酶和抗氧化酶的基因的突变型秀丽隐杆线虫的基因型-表型关系。此外，通过将 CARS 与指纹共焦拉曼分析相结合，我们分析了野生型和突变型秀丽隐杆线虫中脂质的不饱和度。我们的研究表明，在单个活体秀丽隐杆线虫中可以快速定量分析脂质储存、过氧化和去饱和之间的复杂基因型-表型关系。
