Monolayer-fluorescence counting for ultrasensitive detection of tumour cell-derived extracellular vesicles using a step-wedge microfluidic platform

IF 8 1区 化学 Q1 CHEMISTRY, ANALYTICAL Sensors and Actuators B: Chemical Pub Date : 2024-10-17 DOI:10.1016/j.snb.2024.136786
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Abstract

Extracellular vesicles have emerged as significant noninvasive tumour biomarkers, and ultrasensitive detection methods are important for early cancer diagnosis. Here, a monolayer-fluorescence counting strategy was developed for the ultrasensitive detection of extracellular vesicles using a microfluidic platform. Extracellular vesicles were specially captured and separated from complex matrix by immune magnetic microbeads (IMBs). With an enhancing acoustic wave microfluidic channel, the magnetic-extracellular vesicles immune complex was adequately mixed and reacted with antibody-modified fluorescence microbeads (IFBs). Using a newly designed step-wedge microfluidic structure, micrometer-size particles were trapped in large numbers with monolayer in the step zone, and the fluidic resistance was reduced through the wedge structure. This detection signal of the fluorescence-labelled immune sandwich complex could be read by counting, avoiding background interference and achieving an ultrasensitive detection with a detection limit of 850 particles/mL. Moreover, this method showed strong reliability and specificity in clinic samples. In addition, a miniaturized detection device was developed based on this microfluidic platform, improving this detection automation.

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利用阶梯楔形微流体平台进行单层荧光计数,超灵敏检测肿瘤细胞衍生的细胞外囊泡
细胞外囊泡已成为重要的非侵入性肿瘤生物标记物,超灵敏检测方法对早期癌症诊断非常重要。在此,我们开发了一种单层荧光计数策略,利用微流控平台对细胞外囊泡进行超灵敏检测。细胞外囊泡被免疫磁性微珠(IMB)从复杂基质中捕获并分离出来。通过增强声波微流体通道,磁性细胞外囊泡免疫复合物与抗体修饰的荧光微珠(IFBs)充分混合并发生反应。利用新设计的阶梯楔形微流体结构,微米大小的颗粒在阶梯区以单层形式被大量截留,并通过楔形结构降低了流体阻力。荧光标记的免疫夹心复合物的检测信号可通过计数读取,避免了背景干扰,实现了超灵敏检测,检测限为 850 颗粒/毫升。此外,这种方法在临床样本中显示出很强的可靠性和特异性。此外,在该微流控平台的基础上还开发了一种微型检测装置,提高了检测的自动化程度。
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来源期刊
Sensors and Actuators B: Chemical
Sensors and Actuators B: Chemical 工程技术-电化学
CiteScore
14.60
自引率
11.90%
发文量
1776
审稿时长
3.2 months
期刊介绍: Sensors & Actuators, B: Chemical is an international journal focused on the research and development of chemical transducers. It covers chemical sensors and biosensors, chemical actuators, and analytical microsystems. The journal is interdisciplinary, aiming to publish original works showcasing substantial advancements beyond the current state of the art in these fields, with practical applicability to solving meaningful analytical problems. Review articles are accepted by invitation from an Editor of the journal.
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