A portable and sensitive DNA-based electrochemical sensor for detecting piconewton-scale cellular forces

IF 5.7 2区 化学 Q1 CHEMISTRY, ANALYTICAL Analytica Chimica Acta Pub Date : 2024-11-04 DOI:10.1016/j.aca.2024.343392
Mahmoud Amouzadeh Tabrizi , Ahsan Ausaf Ali , Murali Mohana Rao Singuru , Lan Mi , Priyanka Bhattacharyya , Mingxu You
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Abstract

Background

Cell-generated forces are a key player in cell biology, especially during cellular shape formation, migration, cancer development, and immune response. The measurement of forces exerted and experienced by cells is fundamental in understanding these mechanosensitive cellular behaviors. While cell-generated forces can now be detected based on techniques like fluorescence microscopy, atomic force microscopy, optical/magnetic tweezers, however, most of these approaches rely on complicated instruments or materials, as well as skilled operators, which could limit their potential broad applications in regular biological laboratories.

Results

A new type of smartphone-based electrochemical sensor is developed here for cellular force measurement. In this system, a double-stranded DNA-based force probe, known as tension gauge tether, is attached to the surface of a gold screen-printed electrode, which is then incorporated into a portable smartphone-based electrochemical device. Cellular force-induced DNA detachment on the sensor surface results in multiple redox reporters to reach the surface of the electrode and generate enhanced electrochemical signals. To further improve the sensitivity, a CRISPR-Cas12a system has also been incorporated to cleave the remaining surface-attached anchor DNA strand. Using integrin-mediated tension as an example, piconewton-scale adhesion forces generated by ≤ 10 HeLa cells could now be reliably detected. Meanwhile, the threshold forces of these electrochemical sensors can also be modularly tuned to detect different levels of cellular forces.

Significance

These novel DNA-based highly sensitive, portable, cost-efficient, and easy-to-use electrochemical sensors can be potentially powerful tools for detecting different cell-generated molecular forces. Functioning as complementary tools with traction force microscopy and fluorescent probes, these electrochemical sensors can be straightforwardly applied in regular biological laboratories for understanding the basic mechanical principles of cell signaling and for developing novel strategies and materials in tissue engineering, regenerative medicine, and cell therapy.

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基于 DNA 的便携式灵敏电化学传感器,用于检测皮牛顿级细胞力
背景细胞产生的力是细胞生物学中的关键因素,尤其是在细胞形状形成、迁移、癌症发展和免疫反应过程中。测量细胞施加和经历的力是了解这些机械敏感性细胞行为的基础。虽然现在可以通过荧光显微镜、原子力显微镜、光学/磁性镊子等技术检测细胞产生的力,但是这些方法大多依赖于复杂的仪器或材料,以及熟练的操作人员,这可能会限制它们在常规生物实验室中的广泛应用。在这一系统中,一个基于 DNA 的双链力探针(称为张力计系绳)被连接到一个金色丝网印刷电极的表面,然后被整合到一个基于智能手机的便携式电化学装置中。由细胞力引起的 DNA 在传感器表面脱落,导致多个氧化还原报告物到达电极表面,产生增强的电化学信号。为了进一步提高灵敏度,还加入了 CRISPR-Cas12a 系统,以切割剩余的表面附着锚 DNA 链。以整合素介导的张力为例,现在可以可靠地检测到≤10 个 HeLa 细胞产生的皮牛顿级粘附力。同时,这些电化学传感器的阈值力也可以进行模块化调整,以检测不同水平的细胞力。 意义这些基于 DNA 的新型电化学传感器灵敏度高、便于携带、成本效益高且易于使用,是检测不同细胞产生的分子力的有力工具。作为牵引力显微镜和荧光探针的补充工具,这些电化学传感器可直接应用于常规生物实验室,用于了解细胞信号传导的基本机械原理,以及开发组织工程、再生医学和细胞疗法方面的新型策略和材料。
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来源期刊
Analytica Chimica Acta
Analytica Chimica Acta 化学-分析化学
CiteScore
10.40
自引率
6.50%
发文量
1081
审稿时长
38 days
期刊介绍: Analytica Chimica Acta has an open access mirror journal Analytica Chimica Acta: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. Analytica Chimica Acta provides a forum for the rapid publication of original research, and critical, comprehensive reviews dealing with all aspects of fundamental and applied modern analytical chemistry. The journal welcomes the submission of research papers which report studies concerning the development of new and significant analytical methodologies. In determining the suitability of submitted articles for publication, particular scrutiny will be placed on the degree of novelty and impact of the research and the extent to which it adds to the existing body of knowledge in analytical chemistry.
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