Centre for Molecular Nanometrology, WestCHEM, Department of Pure and Applied Chemistry, Technology and Innovation Centre, University of Strathclyde, Glasgow G1 1RD, U.K.
Anal Chem. 2022 Jun 28;94(25):8899-8908. doi: 10.1021/acs.analchem.2c00236. Epub 2022 Jun 14.
Hyperspectral stimulated Raman scattering (SRS) microscopy is a powerful imaging modality for the analysis of biological systems. Here, we report the application of -means cluster analysis (KMCA) of multi-wavelength SRS images in the high-wavenumber region of the Raman spectrum as a robust and reliable method for the segmentation of cellular organelles based on the intrinsic SRS spectrum. KMCA has been applied to the study of the endogenous lipid biochemistry of prostate cancer and prostate healthy cell models, while the corresponding SRS spectrum of the lipid droplet (LD) cluster enabled direct comparison of their composition. The application of KMCA in visualizing the LD content of prostate cell models following the inhibition of de novo lipid synthesis (DNL) using the acetyl-coA carboxylase inhibitor, 5-(tetradecyloxy)-2-furoic acid (TOFA), is demonstrated. This method identified a reliance of prostate cancer cell models upon DNL for metabolic requirements, with a significant reduction in the cellular LD content after treatment with TOFA, which was not observed in normal prostate cell models. SRS imaging combined with KMCA is a robust method for investigating drug-cell interactions in a label-free manner.
高光谱受激拉曼散射（SRS）显微镜是一种强大的生物系统分析成像方式。在这里，我们报告了多波长 SRS 图像的均值聚类分析（KMCA）在拉曼光谱的高波数区域的应用，该方法是一种基于 SRS 光谱内在特征对细胞细胞器进行分割的强大而可靠的方法。KMCA 已应用于前列腺癌和前列腺健康细胞模型的内源性脂质生物化学研究，而脂质滴（LD）簇的相应 SRS 光谱能够直接比较它们的组成。KMCA 的应用可以可视化前列腺细胞模型在从头合成脂质（DNL）抑制后 LD 含量的变化，DNL 抑制使用乙酰辅酶 A 羧化酶抑制剂 5-（十四烷氧基）-2-呋喃酸（TOFA）。该方法确定了前列腺癌细胞模型对 DNL 的代谢需求的依赖性，TOFA 处理后细胞内 LD 含量显著降低，而在正常前列腺细胞模型中则没有观察到这种现象。SRS 成像结合 KMCA 是一种用于以非标记方式研究药物-细胞相互作用的强大方法。
