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首页> 外文期刊>BMC Cancer >Non-destructive, label free identification of cell cycle phase in cancer cells by multispectral microscopy of autofluorescence
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Non-destructive, label free identification of cell cycle phase in cancer cells by multispectral microscopy of autofluorescence

机译：非破坏性的，通过多光谱显微镜的自发荧光显微镜测定癌细胞中细胞周期阶段的自由鉴定

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BACKGROUND:Cell cycle analysis is important for cancer research. However, available methodologies have drawbacks including limited categorisation and reliance on fixation, staining or transformation. Multispectral analysis of endogenous cell autofluorescence has been shown to be sensitive to changes in cell status and could be applied to the discrimination of cell cycle without these steps.METHODS:Cells from the MIA-PaCa-2, PANC-1, and HeLa cell lines were plated on gridded dishes and imaged using a multispectral fluorescence microscope. They were then stained for proliferating cell nuclear antigen (PCNA) and DNA intensity as a reference standard for their cell cycle position (G1, S, G2, M). The multispectral data was split into training and testing datasets and models were generated to discriminate between G1, S, and G2?+?M phase cells. A standard decision tree classification approach was taken, and a two-step system was generated for each line.RESULTS:Across cancer cell lines accuracy ranged from 68.3% (MIA-PaCa-2) to 73.3% (HeLa) for distinguishing G1 from S and G2?+?M, and 69.0% (MIA-PaCa-2) to 78.0% (PANC1) for distinguishing S from G2?+?M. Unmixing the multispectral data showed that the autofluorophores NADH, FAD, and PPIX had significant differences between phases. Similarly, the redox ratio and the ratio of protein bound to free NADH were significantly affected.CONCLUSIONS:These results demonstrate that multispectral microscopy could be used for the non-destructive, label free discrimination of cell cycle phase in cancer cells. They provide novel information on the mechanisms of cell-cycle progression and control, and have practical implications for oncology research.
机译：背景：细胞周期分析对于癌症研究很重要。然而，可用的方法具有缺点，包括有限的分类和依赖固定，染色或转化。已显示内源性细胞自发荧光的多光谱分析对细胞状态的变化敏感，并且可以应用于没有这些步骤的细胞周期的辨别。方法：来自MIA-PACA-2，PANK-1和HELA细胞系的细胞用多光谱荧光显微镜镀在网格上并成像。然后将它们染色以增殖细胞核抗原（PCNA）和DNA强度作为其细胞周期位置（G1，S，G2，M）的参考标准。多光谱数据被分成训练，并生成测试数据集和模型以区分G1，S和G2 + + + +ΔM相位细胞。采用标准决策树分类方法，对每条线产生两步系统。结果：患有癌细胞系的精度为68.3％（MIA-PACA-2）至73.3％（HELA），用于区分G1和G2？+？M，69.0％（MIA-PACA-2）至78.0％（PANC1），以区分S来自G2 + + +。突发的多光谱数据显示自动氟化物NADH，FAD和PPIX在阶段之间存在显着差异。类似地，氧化还原比和蛋白质与游离NADH的蛋白质的比例显着受到影响。结论：这些结果表明，多光谱显微镜可用于癌细胞中的细胞周期阶段的非破坏性标记判断。它们提供了关于细胞周期进展和对照机制的新信息，对肿瘤学研究具有实际影响。

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