Single rare cell assays and biomarkers detection using microfluidic nanotechnology

使用微流控纳米技术进行单一稀有细胞测定和生物标志物检测

• 批准号: 216925-2011
• 负责人: Li, Paul
• 金额: $ 4.01万
• 依托单位: Simon Fraser University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2011
• 资助国家: 加拿大
• 起止时间: 2011-01-01 至 2012-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=477827
• 关键词: Single; rare; cell; assays; biomarkers

The microfluidics nanotechnology is employed to enable two streams of research: (1) information-rich assays on single rare cells, and (2) high-throughput microarray-based assays for biomarkers (e.g. nucleic acids and proteins). STREAM 1 is to use a single cell chip to identify and measure single tumor cells contained within a nanolitre-sized cell retention chamber. The measured data may provide information for early disease diagnosis and for prognosis during treatment. Cells such as circulating tumor cells (CTCs) found in blood samples are rare. Our single cell biochip is able to measure a rare cell for multiple cellular parameters, such as cell size, intracellular drug concentration, and cell surface biomarker for multidrug resistance (MDR). We will perform drug accumulation measurements on multiple single cells using MDR inhibitors in order to enhance accumulation; we expect to correlate these measurement results with the immunoassay-based MDR biomarker determination. The measured data of drug accumulation on a single cancer cell with or without the MDR inhibitor may provide information for effective chemotherapy such as the use of the appropriate drug-inhibitor combination. STREAM 2 is to conduct analysis of DNA and proteins biomarkers for high throughput diagnosis. The analysis will be conducted on a microfluidic microarray (MMA) chip developed for fungal pathogen detection. We propose to detect DNA biomarkers on the MMA chip, and to increase detection specificity enhanced by using gold nanoparticles. One application is to detect diseases related to salmon health. The MMA chip is also employed to analyze protein biomarkers in human serum samples for disease detection and treatment. We will also perform computer simulation study of the microfluidic liquid flow process for efficient reagent/sample delivery. Both platforms on single rare cell assays and MMA are suitable for incorporation in point-of-care (POC) systems. The novelty of all projects involves the efficient manipulation of inherently low liquid volumes (in sub-nanolitre) within these microfluidic devices, thus demanding less starting materials and generating less wastes, conforming to the notion of green chemistry.

微流控纳米技术被用于两个研究流：(1)针对单个稀有细胞的信息丰富的分析；(2)基于高通量微阵列的生物标记物(如核酸和蛋白质)的分析。Stream 1是使用单个细胞芯片来识别和测量纳升大小的细胞保留室中包含的单个肿瘤细胞。测量数据可为疾病的早期诊断和治疗过程中的预后提供信息。在血液样本中发现的循环肿瘤细胞(CTCs)等细胞很少见。我们的单细胞生物芯片能够测量罕见细胞的多个细胞参数，如细胞大小、细胞内药物浓度和细胞表面多药耐药性(MDR)生物标志物。我们将使用多药耐药抑制剂在多个单细胞上进行药物积累测量，以促进药物积累；我们希望将这些测量结果与基于免疫分析的多药耐药生物标志物测定相关联。使用或不使用MDR抑制剂的单个癌细胞上药物累积的测量数据可能为有效的化疗提供信息，例如使用适当的药物-抑制剂组合。Stream 2用于高通量诊断的DNA和蛋白质生物标记物的分析。分析将在为检测真菌病原体而开发的微流控微阵列(MMA)芯片上进行。我们建议在MMA芯片上检测DNA生物标记物，并通过使用金纳米颗粒来提高检测特异性。其中一个应用是检测与鲑鱼健康相关的疾病。MMA芯片还被用于分析人类血清样本中的蛋白质生物标记物，用于疾病检测和治疗。我们还将对微流控液体流动过程进行计算机模拟研究，以实现高效的试剂/样品输送。基于单一稀有细胞分析和MMA的两个平台都适合用于医疗点(POC)系统。所有项目的新颖性都涉及有效地处理这些微流控装置内固有的低液体体积(以亚纳升为单位)，因此需要更少的起始材料和产生更少的废物，这符合绿色化学的概念。