Abstract

A fast and convenient method of recognizing the tongue tumor tissue based on bioelectrical impedance spectroscopy (BIS) is proposed. According to the difference among the electrical characteristics of tongue tissue under different pathological and physiological conditions, we can judge whether it is pathological. This method can help the surgeon in the clinical resection of tongue cancer not only to remove the tumor completely, but also to retain the patient’s tongue function as much as possible. In this paper, a model of human tongue squamous cell carcinoma (HSC3) xenografted <i>in situ</i> is established in mice. The electrical properties of the normal tissue, mixed tumor tissue and tumor tissue are studied by the BIS technology. The amplitude spectrum shows that none of the three tissues can be distinguished well in a low frequency range of 100–8.09 × 10<sup>5</sup> Hz due to the influence of contact impedance, but they can be distinguished according to their electrical characteristics in a high frequency range of 8.09 × 10<sup>5</sup>–5 × 10<sup>6</sup> Hz. In the process of the experiment, first of all, the tip, middle and root of the normal tongue are detected, and the results show that the impedance values of these three parts are similar in the high frequency band, so the influence of different positions of tongue on the impedance value can be excluded. Then, the same three regions of the cancerous tongue are detected, and three electrical parameters, namely relaxation frequency <i>f</i><sub>relax</sub>, real part spectrum <inline-formula><tex-math id="M3">\begin{document}${{{Z}}'_{\rm{relax}}}$\end{document}</tex-math><alternatives><graphic xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="15-20210297_M3.jpg"/><graphic xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="15-20210297_M3.png"/></alternatives></inline-formula> and imaginary part spectrum <inline-formula><tex-math id="M4">\begin{document}${{{Z}}''_{\rm{relax}}}$\end{document}</tex-math><alternatives><graphic xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="15-20210297_M4.jpg"/><graphic xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="15-20210297_M4.png"/></alternatives></inline-formula> of electrical impedance are extracted from the amplitude spectrum of high frequency band. The quantitative analyses of these three kinds of tissues show that the <inline-formula><tex-math id="M5">\begin{document}${{{Z}}'_{\rm{relax}}}$\end{document}</tex-math><alternatives><graphic xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="15-20210297_M5.jpg"/><graphic xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="15-20210297_M5.png"/></alternatives></inline-formula> and <inline-formula><tex-math id="M6">\begin{document}${{{Z}}''_{\rm{relax}}}$\end{document}</tex-math><alternatives><graphic xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="15-20210297_M6.jpg"/><graphic xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="15-20210297_M6.png"/></alternatives></inline-formula> of tumor tissue are the highest, and those of normal tissue are the lowest. Finally, the cancerous tissue coefficients <i>α</i> and <i>β</i> (the relative change percentage of the real part and imaginary part of the impedance value between the tested tissue and normal tissue) are determined according to these three electrical parameters for tumor tissue identification. The results show that when <i>α</i> ≤ 36.5% and <i>β</i> ≤ 31.2%, the tissue is normal; when <i>α</i> ≥ 36.5% and <i>β</i> ≥ 31.2%, the tissue may be mixed with tumor tissue; when <i>α</i> ≥ 82.7% and <i>β</i> ≥ 73.6%, the tissue is tumor tissue.

Highlights

  • In the process of the experiment, first of all, the tip, middle and root of the normal tongue are detected, and the results show that the impedance values of these three parts are similar in the high frequency band, so the influence of different positions of tongue on the impedance value can be excluded

  • The results show that when a ≤ 36.5% and b ≤ 31.2%, the tissue is normal; when a ≥ 36.5% and b ≥ 31.2%, the tissue may be mixed with tumor tissue; when a ≥ 82.7% and b ≥ 73.6%, the tissue is tumor tissue

  • 1) (College of Electrical and Mechanical Engineering, Nanjing University of Aeronautics and Astronautics, Nanjing 210016, China) 2) (Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing 210008, China) ( Received 8 February 2021; revised manuscript received 19 March 2021 )

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Summary

HSC3 舌体肿瘤制备

HSC3 细胞培养使用了 10% 胎牛血清、100 U/mL 青霉 素和 100 mg/mL 链霉素的 DMEM 培养基, 置于 37 °C、5% CO2 的恒温培养箱中培养. 准备 4 只 Balb/c-nude 雌性小鼠, 当细胞培养完成后, 在其 中 3 只小鼠麻醉状态下注射 1 × 106 细胞数量的 HSC3 细胞于每只小鼠的舌尖部位. 每日观察小鼠 舌尖并用游标卡尺测量肿瘤的长和宽, 然后用公式 (V = (L × W 2)/2 , 其中 L 是肿瘤的长度, W 是肿 瘤的宽度) 计算其大致体积. 待肿物形成后, 用颈 椎脱臼法处死麻痹状态下的小鼠, 最后切取小鼠的 舌体置于磷酸缓冲盐 (PBS) 溶液中以便备用. 准备两个培养皿, 用镊子将 4 只小鼠的舌体夹 入其中 1 个培养皿以待检测, 并往培养皿中倒入 PBS 溶液以保持舌组织的活性. 图 1 BIS 检测仪器 (a) 阻抗分析仪及传感器; (b) 四电极传感器结构图 Fig. 1. Measuring equipment of BIS: (a) Impedance analyzer and sensor; (b) structure diagram of four-electrode sensor. 沿着舌体中线进行切片处理, 切片组织用 95% 乙 醇 固 定 20 min, 之 后 用 PBS 溶 液 洗 涤 , 每 次 1 min. 分色之后 用伊红染液对贴片组织中细胞的细胞质染色, 染 色 1 min 后用流水冲洗. 沿着舌体中线进行切片处理, 切片组织用 95% 乙 醇 固 定 20 min, 之 后 用 PBS 溶 液 洗 涤 , 每 次 1 min. 洗涤后用苏木素染液对切片组织中细胞的 细胞核染色, 染色 2—3 min 后用流水冲洗. 然后 通过显微镜观察, 如果细胞核染色过深, 用 1% 盐 酸酒精溶液分色数秒, 之后用流水冲洗. 分色之后 用伊红染液对贴片组织中细胞的细胞质染色, 染 色 1 min 后用流水冲洗. 最后自然晾干切片组织并 用中性树胶封片

HE 染色结果
BIS 检测结果
HSC3 癌变舌体中不同区域组织的定量 分析
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