Broadband Electrical Sensing of Nuclear Morphology and DNA Content in a Single Live Cell

需要以紧凑和非侵入性方式感测细胞内特性（例如核特征）的新方法来提高癌症诊断的速度和准确性。这种方法还将实现对细胞核的实时动态监测，这可以有助于对细胞发育和恶性进展的基本理解。目前，核形态和DNA含量的检测通过光学显微镜和流式细胞术，这两个都是庞大的，微妙的，需要细胞标记。为了满足便携式和廉价的癌症筛查技术的需求，宽带电感测在这里提出了从单个细胞中提取有关核形态和DNA含量的信息。拟议的工作是建立在研究小组的工作在宽带电感测单细胞内的电阻和电容。所提出的电传感器可以很容易地与芯片实验室系统集成，使其能够在护理点广泛使用，这将对医疗保健产生更广泛的影响。在不物理穿透细胞膜的情况下监测细胞内细胞器的能力不仅将有助于癌症细胞学，而且还将允许实时监测核动力学，这可以改变对细胞发育，癌症治疗和细胞生物学的许多其他方面的研究。通过课堂教学，会议，出版物和其他推广工作，从这项研究的知识将传播给学生，专业人士和公众。该研究将允许参与K-12，本科生和研究生，特别是女性和少数民族学生的多学科培训。PI和co-PI在招募代表性不足的学生进行研究和研究生教育方面有着良好的记录。拟议的研究旨在建立一个微波等效的共聚焦显微镜用于单细胞深度分析，揭示细胞内的细节，如核形态和DNA含量的变化。这项工作是基于这样的假设，即宽带电感测可以揭示具有不同复介电常数和弛豫频率的不同细胞内隔室的介电性质。1）设计并制作出灵敏度和空间分辨率均满足9 kHz ~ 9 GHz蜂窝内宽带电学传感要求的多端口共面波导。2)建立一个多尺度模型，以了解电场与细胞膜、细胞质和细胞核的相互作用，包括细胞核大小、形状和位置以及DNA含量的变化。3)对单个活的人类细胞进行宽带电感测和信号分析，以验证多尺度模型并提取核形态和DNA含量。这项研究的知识将提供电场如何在广泛的频率范围内与细胞及其细胞器相互作用的基本理解。宽带电生物传感器和多尺度模型将被开发，以允许具有高空间分辨率、信噪比、通量和再现性的细胞内隔室的介电表征，这将广泛影响生物工程和电气工程，特别是生物电子学、生物物理学、该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的知识产权进行评估来支持。优点和更广泛的影响审查标准。

New approaches to sense intra-cellular properties such as nuclear characteristics in a compact and non-invasive fashion are needed to increase the speed and accuracy of cancer diagnosis. Such approaches will also enable real-time dynamic monitoring of a cell nucleus, which can contribute to fundamental understanding of cell development and malignancy progression. Currently, nuclear morphology and DNA content are inspected through optical microscopy and flow cytometry, both are bulky, delicate and require cell labeling. To satisfy the need of portable and inexpensive technologies for cancer screening, broadband electrical sensing is proposed here to extract information about nuclear morphology and DNA content from a single cell. The proposed work is built on the research team's work in broadband electrical sensing of single-cell intra-cellular resistance and capacitance. The proposed electrical sensor can be readily integrated with a lab-on-chip system to enable its wide-spread use at the point of care, which will have broader impacts on healthcare. The capability to monitor intra-cellular organelles without physically penetrating a cell membrane not only will contribute to cancer cytology, but also will permit real-time monitoring of nuclear dynamics, which can transform the research on cell development, cancer therapeutics, and many other aspects of cell biology. Through classroom teaching, conferences, publications, and other outreach efforts, knowledge from this research will be disseminated to students, professionals, and general public. The research will allow multi-disciplinary training to participating K-12, undergraduate and graduate students, especially female and minority students. The PI and co-PI have strong track records in recruiting underrepresented students to research and graduate education.The proposed research intends to build a microwave equivalent of confocal microscopy for single cell depth profiling, revealing intracellular details such as alterations in nuclear morphology and DNA content. This work is based on the hypothesis that broadband electrical sensing can reveal dielectric properties of different intra-cellular compartments that have distinct complex permittivities and relaxation frequencies. The following aims will be achieved: 1) Design and fabricate multi-port coplanar waveguides with sensitivity and spatial resolution meeting the needs for intra-cellular broadband electrical sensing from 9 kHz to 9 GHz. 2) Develop a multi-scale model to understand the interaction of electric fields with cell membrane, cytoplasm and nucleus, including variation of the size, shape and location of the nucleus as well as its DNA content. 3) Perform broadband electrical sensing and signal analysis of single live human cells to validate the multi-scale model and to extract nuclear morphology and DNA content. Knowledge from this research will provide fundamental understanding of how electric fields interact with a cell and its organelles in a broad range of frequencies. Broadband electrical biosensors and multi-scale models will be developed to allow dielectric characterization of intra-cellular compartments with high spatial resolution, signal-to-noise ratio, throughput and reproducibility, which will broadly impact both bioengineering and electrical engineering, especially bioelectronics, biophysics, cancer biology and cell biology.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.