Autonomous magnetically actuated continuous flow microimmunofluorocytometry assay.
Autonomous magnetically actuated continuous flow microimmunofluorocytometry assay.
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DOI：
10.1007/s10404-009-0543-1
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发表时间：
2010-08-01
影响因子：
2.8
通讯作者：
Zahn, Jeffrey D.
中科院分区：
文献类型：
--
作者：
Sasso, Lawrence A.;Undar, Akif;Zahn, Jeffrey D.
本文提出了一种微流控装置，它集成了连续流动环境中的细胞计数珠的自主系列免疫荧光结合反应与荧光检测和定量。微设备检测旨在减轻进行传统免疫检测时使用的大量实验室工作和大量样品，而无需复杂的外部质控品。该技术基于抗原夹心免疫测定的系列处理步骤的小型化和自动化，使用顺磁性微珠进行集成荧光检测。连续流设计可以使得能够对用于临床应用的连续输注的样品中的时变蛋白质浓度进行时间跟踪，特别是用于监测在涉及心肺转流（CPB）程序的心脏手术期间产生的血液中的炎症标志物蛋白。首先通过单个孵育装置验证装置操作，该孵育装置使用链霉亲和素包被的顺磁性细胞计数珠测量荧光标记的生物素分子的浓度。随后，使用过敏毒素补体蛋白C3 a样本对双孵育装置进行了测试，结果表明，双孵育装置能够区分蛋白质典型全身浓度（1-5 μg/ml）的样本，样本使用量非常低（<6 μl/h）。据信，这种连续流动的自动化微量免疫传感器技术将成为高采样率免疫测定的平台，能够跟踪和更彻底地表征全身炎症过程，并可能有助于开发CPB期间和之后全身炎症的更好治疗方案。
This article presents a microfluidic device which integrates autonomous serial immunofluorocytometry binding reactions of cytometric beads with fluorescence detection and quantification in a continuous flow environment. The microdevice assay is intended to alleviate the extensive benchwork and large sample volumes used when conducting traditional immunoassays, without requiring complex external controls. The technology is based on the miniaturization and automation of the serial processing steps of an antigen sandwich immunoassay, with integrated fluorescence detection using paramagnetic microbeads. The continuous flow design may enable temporal tracking of time-varying protein concentrations in a continuously infused sample for clinical applications, specifically for monitoring inflammation marker proteins in blood produced during cardiac surgeries involving cardiopulmonary bypass (CPB) procedures. The device operation was first validated via a single incubation device which measured the concentration of a fluorescently labeled biotin molecule using streptavidin-coated paramagnetic cytometric beads. Subsequently, a dual incubation device was tested with samples of the anaphylatoxin complement protein C3a, and was shown to be capable of differentiating between samples at typical systemic concentrations of the protein (1–5 μg/ml), with very low sample usage (<6 μl/h). It is believed that this continuous flow, automated microimmunosensor technology will be a platform for high sample rate immunoassays capable of tracking and more thoroughly characterizing the systemic inflammation process, and may aid in the development of better treatment options for systemic inflammation during and after CPB.
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