Development of a label-free, impedance-based biosensor to identify harmful effects of pesticides on insect cells

Sandra Friedrich, Neha Malagimani, Stefanie Michaelis, Joachim Wegener
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Abstract

Insects are a major part of the planet's ecosystem and their vital role as pollinators for agriculture is undisputed. Alongside factors as climate change or loss of habitats, rising use of pesticides emerges as a key threat to insect populations. For fighting this man-made problem, development of an easy, fast, sensitive, and non-invasive biosensor for determining pesticide toxicity may help to ban harmful substances and formulations. Here, a biosensor based on Sf21 (Spodoptera frugiperda) insect cells as sensors and electric cell-substrate impedance sensing (ECIS) as physical transducer is described. Sf21 cell suspensions and well-defined pesticide solutions were mixed immediately before seeding on planar gold-film electrodes. The capacitance at 20 kHz was recorded as a function of time as a measurand for cell adhesion providing dose–response profiles of pesticide impact. For future in-field applications, decoupling of the cell culture routines from the actual cytotoxicity assay is mandatory. Thus, suspensions of Sf21 cells were cryopreserved at –80°C in the wells of multielectrode arrays and thawed anytime for conducting the assays. Five pesticides were tested for their concentration-dependent cytotoxicity expressed as EC50 values by ECIS and validated using the well-established WST-1 cell viability assay. Results were found to be in good agreement. Our studies revealed cytotoxic effects of some pesticides sold for home usage far below the recommended concentration and were found to be more toxic than formulations sold for agricultural industry only.

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开发无标记、基于阻抗的生物传感器,以识别农药对昆虫细胞的有害影响
昆虫是地球生态系统的重要组成部分,它们作为农业授粉者的重要作用毋庸置疑。除气候变化或栖息地丧失等因素外,杀虫剂使用量的增加也成为昆虫种群面临的主要威胁。为了解决这一人为问题,开发一种简便、快速、灵敏和非侵入性的生物传感器来确定杀虫剂的毒性,可能有助于禁止使用有害物质和制剂。本文介绍了一种以 Sf21(Spodoptera frugiperda)昆虫细胞为传感器、以电细胞-基底阻抗传感(ECIS)为物理换能器的生物传感器。将 Sf21 细胞悬浮液和定义明确的杀虫剂溶液混合后立即播种到平面金膜电极上。记录 20 kHz 频率下的电容与时间的函数关系,作为细胞粘附的测量值,提供杀虫剂影响的剂量反应曲线。对于未来的实地应用,必须将细胞培养程序与实际的细胞毒性检测解耦。因此,将 Sf21 细胞悬浮液冷冻保存在零下 80 摄氏度的多电极阵列孔中,并随时解冻以进行检测。用 ECIS 测试了五种农药的浓度依赖性细胞毒性(以 EC50 值表示),并用成熟的 WST-1 细胞活力测定法进行了验证。结果表明,这五种农药的浓度依赖性细胞毒性与 ECIS 的 EC50 值非常吻合。我们的研究揭示了一些家用杀虫剂的细胞毒性效应远远低于推荐浓度,而且比仅用于农业的制剂毒性更大。本文受版权保护。
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