Issadore David
University of Pennsylvania, 210 South 33rd Street, Suite 240 Skirkanich Hall, Philadelphia, PA, 19104-6321, USA,
Methods Mol Biol. 2015;1256:123-37. doi: 10.1007/978-1-4939-2172-0_9.
The sparse cells that are shed from tumors into peripheral circulation are an increasingly promising resource for noninvasive monitoring of cancer progression, early diagnosis of disease, and serve as a tool for improving our understanding of cancer metastasis. However, the extremely sparse concentration of circulating tumor cells (CTCs) in blood (~1-100 CTC in 7.5 mL of blood) as well as their heterogeneous biomarker expression has limited their detection using conventional laboratory techniques. To overcome these challenges, we have developed a microfluidic chip-based micro-Hall detector (μHD), which can directly measure single, immunomagnetically tagged cells in whole blood. The μHD can detect individual cells even in the presence of vast numbers of blood cells and unbound reactants, and does not require any washing or purification steps. Furthermore, this cost-effective, single-cell analytical technique is well suited for miniaturization into a mobile platform for low-cost point-of-care use. In this chapter, we describe the methodology used to design, fabricate, and apply these chips to cancer diagnostics.
从肿瘤脱落进入外周循环的稀疏细胞,对于癌症进展的无创监测、疾病的早期诊断而言,是一种越来越有前景的资源,并且可作为一种工具,帮助我们更好地理解癌症转移。然而,血液中循环肿瘤细胞(CTC)的浓度极低(每7.5毫升血液中约有1 - 100个CTC),以及它们异质性的生物标志物表达,限制了使用传统实验室技术对其进行检测。为了克服这些挑战,我们开发了一种基于微流控芯片的微霍尔探测器(μHD),它可以直接在全血中测量单个免疫磁珠标记的细胞。μHD即使在存在大量血细胞和未结合反应物的情况下也能检测单个细胞,并且不需要任何洗涤或纯化步骤。此外,这种具有成本效益的单细胞分析技术非常适合小型化到移动平台,用于低成本的即时检测。在本章中,我们描述了用于设计、制造这些芯片并将其应用于癌症诊断的方法。