Faculty of Biomedical Engineering, Czech Technical University, nam. Sitna 3105, 272 01 Kladno, Czech Republic.
Institute of Chemical Process Fundamentals of the ASCR, Rozvojova 135, 165 00 Prague, Czech Republic.
Biosensors (Basel). 2023 Feb 11;13(2):256. doi: 10.3390/bios13020256.
In this paper, we analysed the steady state fluorescence spectra of cell suspensions containing healthy and carcinoma fibroblast mouse cells, using a genetic-algorithm-spectra-decomposition software (GASpeD). In contrast to other deconvolution algorithms, such as polynomial or linear unmixing software, GASpeD takes into account light scatter. In cell suspensions, light scatter plays an important role as it depends on the number of cells, their size, shape, and coagulation. The measured fluorescence spectra were normalized, smoothed and deconvoluted into four peaks and background. The wavelengths of intensities' maxima of lipopigments (LR), FAD, and free/bound NAD(P)H (AF/AB) of the deconvoluted spectra matched published data. In deconvoluted spectra at pH = 7, the fluorescence intensities of the AF/AB ratio in healthy cells was always higher in comparison to carcinoma cells. In addition, the AF/AB ratio in healthy and carcinoma cells were influenced differently by changes in pH. In mixtures of healthy and carcinoma cells, AF/AB decreases when more than 13% of carcinoma cells are present. Expensive instrumentation is not required, and the software is user friendly. Due to these attributes, we hope that this study will be a first step in the development of new cancer biosensors and treatments with the use of optical fibers.
在本文中，我们使用遗传算法光谱分解软件（GASpeD）分析了含有健康和癌性成纤维细胞的小鼠细胞悬浮液的稳态荧光光谱。与其他解卷积算法（如多项式或线性解混软件）不同，GASpeD 考虑了光散射。在细胞悬浮液中，光散射起着重要作用，因为它取决于细胞数量、大小、形状和凝聚。测量的荧光光谱经过归一化、平滑和解卷积为四个峰和背景。解卷积光谱中脂色素（LR）、FAD 和游离/结合 NAD(P)H（AF/AB）的强度最大值波长与已发表的数据相匹配。在 pH = 7 时的解卷积光谱中，与癌细胞相比，健康细胞的 AF/AB 比值的荧光强度总是更高。此外，pH 值变化对健康细胞和癌细胞的 AF/AB 比值的影响不同。在健康细胞和癌细胞的混合物中，当存在超过 13%的癌细胞时，AF/AB 会降低。不需要昂贵的仪器，并且软件易于使用。由于这些属性，我们希望这项研究将是开发使用光纤的新型癌症生物传感器和治疗方法的第一步。
