Electronic Microflow Cytometry for Surface Expression Analysis of Tumor Cells

用于肿瘤细胞表面表达分析的电子微流式细胞仪

• 批准号: 1610995
• 负责人: Ali Fatih Sarioglu
• 金额: $ 40万
• 依托单位: Georgia Tech Research Corporation
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1610995&HistoricalAwards=false
• 关键词: Electronic; Microflow; Cytometry; Surface; Expression

Electronic Microflow Cytometry for Surface Expression Analysis of Cancer CellsProposal Objectives:Cellular heterogeneity is the hallmark of cancer with individual cancer cells differing in their tumor initiating potential and therapeutic resistance. Tumor cell heterogeneity is often obscured by population level measurements in systems biology and medicine; yet it has significant implications in disease progression. This proposal is about bio-analytical technologies that are capable of analyzing individual cancer cells and offer exciting opportunities for early-detection and monitoring of cancer while it is still manageable, and for gaining biological insight to develop effective therapies. Non-Technical Abstract:While capable of analyzing single cells in detail, current technologies are exclusively limited to large well-equipped laboratories due to their high cost, operational complexity and bulky instrumentation. This proposal will address these shortcomings by developing a fundamentally different single cell analysis platform that is fast, low-cost, portable and hence more amenable to point-of-care use than conventional technologies. To achieve this, we will develop a lab-on-a-chip platform that integrates microfluidic cell sorting with electronic sensing by utilizing engineering approaches from multiple technical disciplines including microfluidics, electronics and telecommunications. To further enhance the broader impact of this program, we will be involved with outreach activities to (1) attract K-12 and high school students to STEM education by engaging with local and elementary schools, (2) engage undergraduate students in research activities, (3) attract underrepresented students in engineering by utilizing Georgia Tech's diverse student body, (4) promote teaching and training by contributing to teacher education programs, (5) disseminate the results of the proposed research program to the society through media releases and lab website.Technical Abstract:Flow cytometry is an invaluable technique for comprehensive biophysical and biochemical analysis of single cells. In flow cytometry, single cells in a fluid stream are interrogated one by one and subsequently sorted based on the measured properties. The objective of this proposal is to integrate inherently parallel microfluidic cell sorting with a parallel electronic sensor array to create a fundamentally different flow cytometer. The research tasks to be undertaken as part of this project are (1) to develop a scalable electronic sensor that utilizes resistive pulse sensing to simultaneously detect particles in multiple microfluidic channels from a single electrical output, (2) to design a microfluidic cell sorter based on magnetophoresis and integrate it with the developed electronic sensor array to quantify sorted cells, (3) to analyze surface expression of cancer cells by magnetic pre-labeling and benchmark against established laboratory techniques. In this proposal, we combine two traditionally distinct technical disciplines, microfluidics and telecommunications to design a scalable electronic readout mechanism for interfacing microfluidic chips. Our technology relies on multiplexing an array of micromachined Coulter counters each designed to produce a distinct digital code when a particle is detected. These digital codes are developed using the same principles of Code Division Multiple Access (CDMA) telecommunication networks and can be uniquely recovered through simple mathematical calculations. This approach will be used to increase number of sensors without increasing the hardware complexity. Instead, complexity in the hardware will be shifted towards computational algorithms to decode the signal. We will then integrate this sensor with magnetophoretic sorting of tumor cells to quantify the expression of a protein with clinical utility for managing and guiding cancer therapy.

目的：肿瘤细胞的异质性是肿瘤细胞的特征，不同的癌细胞具有不同的致瘤潜能和治疗耐药性。在系统生物学和医学中，肿瘤细胞的异质性常常被种群水平的测量所掩盖；然而，它在疾病进展中有着重要的意义。这项提议是关于生物分析技术，这些技术能够分析单个癌细胞，并提供令人兴奋的机会，在癌症仍可控制的情况下及早发现和监测，并获得生物学洞察力以开发有效的治疗方法。非技术摘要：虽然目前的技术能够详细分析单个细胞，但由于其成本高、操作复杂和仪器庞大，目前的技术仅限于设备齐全的大型实验室。这项提议将通过开发一种根本不同的单细胞分析平台来解决这些缺点，该平台快速、低成本、便携，因此比传统技术更适合使用护理点。为了实现这一目标，我们将开发一个芯片实验室平台，利用微流体、电子学和电信等多个技术学科的工程方法，将微流控细胞分类与电子传感相结合。为了进一步增强这一计划的广泛影响，我们将参与下列外联活动：(1)通过与当地和小学的接触吸引K-12和高中生接受STEM教育，(2)让本科生参与研究活动，(3)利用佐治亚理工学院多样化的学生群体吸引未被充分代表的学生进入工程学，(4)通过促进教师教育计划促进教学和培训，(5)通过媒体发布和实验室网站向社会传播拟议研究计划的结果。在流式细胞术中，流体中的单个细胞被一个接一个地询问，然后根据测量的特性进行分类。这项提议的目标是将本质上并行的微流控细胞分选与并行电子传感器阵列相结合，以创建一种完全不同的流式细胞仪。作为该项目的一部分，将开展的研究任务包括：(1)开发一种可扩展的电子传感器，利用阻性脉冲传感技术从单个电子输出端同时检测多个微流控通道中的颗粒；(2)设计一种基于磁感应原理的微流控细胞分选机，并将其与开发的电子传感器阵列集成，对分选后的细胞进行量化；(3)通过磁性预标记分析癌细胞的表面表达，并以既定的实验室技术为基准。在这个方案中，我们结合了微流体学和电信这两个传统上截然不同的技术学科来设计一种可扩展的电子读出机制，用于连接微流控芯片。我们的技术依赖于多路复用一组微机械库尔特计数器，每个计数器都被设计成在检测到粒子时产生不同的数字代码。这些数字代码是使用码分多址(CDMA)电信网络的相同原理开发的，并且可以通过简单的数学计算唯一地恢复。这种方法将用于在不增加硬件复杂性的情况下增加传感器的数量。取而代之的是，硬件的复杂性将转移到解码信号的计算算法上。然后，我们将把这种传感器与肿瘤细胞的磁感应分选相结合，以量化具有临床实用价值的蛋白质的表达，以管理和指导癌症治疗。