Phenotype Identification of HeLa Cells Knockout CDK6 Gene Based on Label-Free Raman Imaging,Analytical Chemistry

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Phenotype Identification of HeLa Cells Knockout CDK6 Gene Based on Label-Free Raman Imaging
Analytical Chemistry ( IF 6.7 ) Pub Date : 2022-06-15 , DOI: 10.1021/acs.analchem.2c00188
Ying Xu ₁ , Xiaoli Hou ₂ , Qiaoqiao Zhu ₁ , Shijie Mao ₁ , Jie Ren ₁ , Jidong Lin ₁ , Ning Xu ₁

Affiliation

Identifying cell phenotypes is essential for understanding the function of biological macromolecules and molecular biology. We developed a noninvasive, label-free, single-cell Raman imaging analysis platform to distinguish between the cell phenotypes of the HeLa cell wild type (WT) and cyclin-dependent kinase 6 (CDK6) gene knockout (KO) type. Via large-scale Raman spectral and imaging analysis, two phenotypes of the HeLa cells were distinguished by their intrinsic biochemical profiles. A significant difference was found between the two cell lines: large lipid droplets formed in the knockout HeLa cells but were not observed in the WT cells, which was confirmed by Oil Red O staining. The band ratio of the Raman spectrum of saturated/unsaturated fatty acids was identified as the Raman spectral marker for HeLa cell WT or gene knockout type differentiation. The interaction between organelles involved in lipid metabolism was revealed by Raman imaging and Lorentz fitting, where the distribution intensity of the mitochondria and the endoplasmic reticulum membrane decreased. At the same time, lysosomes increased after the CDK6 gene knockout. The parameters obtained from Raman spectroscopy are based on hierarchical cluster analysis and one-way ANOVA, enabling highly accurate cell classification.

中文翻译：

基于无标记拉曼成像的HeLa细胞敲除CDK6基因表型鉴定

识别细胞表型对于了解生物大分子和分子生物学的功能至关重要。我们开发了一种无创、无标记、单细胞拉曼成像分析平台，以区分 HeLa 细胞野生型 (WT) 和细胞周期蛋白依赖性激酶 6 (CDK6) 基因敲除 (KO) 型的细胞表型。通过大规模拉曼光谱和成像分析，HeLa 细胞的两种表型通过其内在的生化特征来区分。发现两种细胞系之间存在显着差异：在敲除的 HeLa 细胞中形成大脂滴，但在 WT 细胞中未观察到，这通过油红 O 染色得到证实。饱和/不饱和脂肪酸的拉曼光谱的带比被确定为HeLa细胞WT或基因敲除型分化的拉曼光谱标记。拉曼成像和洛伦兹拟合揭示了参与脂质代谢的细胞器之间的相互作用，其中线粒体和内质网膜的分布强度降低。同时，CDK6基因敲除后溶酶体增加。从拉曼光谱获得的参数基于层次聚类分析和单向方差分析，可实现高度准确的细胞分类。CDK6基因敲除后溶酶体增加。从拉曼光谱获得的参数基于层次聚类分析和单向方差分析，可实现高度准确的细胞分类。CDK6基因敲除后溶酶体增加。从拉曼光谱获得的参数基于层次聚类分析和单向方差分析，可实现高度准确的细胞分类。

更新日期：2022-06-15

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