An integrated lab-on-a-disc for automated cell-based allergen screening bioassays
An integrated lab-on-a-disc for automated cell-based allergen screening bioassays
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DOI：
10.1016/j.talanta.2012.03.061
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发表时间：
2012-08-15
期刊：
影响因子：
6.1
通讯作者：
Ho, H. P.
中科院分区：
文献类型：
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作者：
Chen, Q. L.;Cheung, K. L.;Ho, H. P.
我们利用各种阀门方案来利用纯粹的旋转调节流量控制，以在基于离心机的磁盘实验室（LOAD）上实现全面的基于细胞的生物测定（CBB）。首次开发了LOAD来检查活嗜碱性粒细胞的过敏性脱颗粒以进行过敏原筛查，还可以对其进行调整以适应广泛的CBB。在该系统中，控制过敏反应连同介质分离嗜碱性粒细胞使用虹吸阀和离心沉降内实现微结构网络。整个脱颗粒分析过程包括按需释放样品、反应和脱颗粒、过敏介质分离和检测，在单次运行中以自动顺序执行。为了验证我们的基于细胞的方法，检测脱粒介导的已知分泌，离子霉素或趋化肽甲酰甲硫氨酸亮氨酸苯丙氨酸（fMLP），首先证明。使用真实的过敏原屋尘螨蛋白（Der p1）及其相应的人血清IgE的进一步实验也显示阳性结果。该测定的总效率为80.6%，与其他常规方法相当。该设备在一个磁盘上以并行格式运行4个相同的单元，提供了单步多路过敏原筛查的可能性。该设备能够在30分钟内报告结果。它具有许多理想的优点，包括快速和多路分析，成本低，单步操作，最小的样本量，更少的不适，最重要的是增加了安全性，因为患者不再容易受到常见的皮肤点刺试验引起的过敏性休克反应。设备内流量控制的灵活性使其适用于各种CBB。(C)2012爱思唯尔有限公司版权所有。
We have utilized various valving scheme to leverage purely rotation-regulated flow control to enable comprehensive cell-based bioassays (CBBs) on centrifuge-based lab-on-a-disc (LOAD). A LOAD has been developed to examine allergic degranulation from live basophils for allergens screening for the first time, which can also be adjusted to suit a wide range of CBBs. In this system, controlled allergic reaction together with mediator separation from basophils using siphon valving and centrifugal sedimentation are realized inside microstructured network. The entire degranulation analysis process including on-demand release of samples, reaction and degranulation, allergic mediator separation and detection is executed in an automatic sequence within a single run. To validate our cell-based approach, detection of degranulation mediated by known secretagagues, ionomycin or chemotatic peptide formyl-methionine-leucine-pheylalanine (fMLP), is first demonstrated. Further experiments using real allergens house dust mite protein (Der p1) and its corresponding human serum IgE also show positive results. The overall efficiency of the assay is 80.6%, which is comparable to other conventional methods. With 4 identical units on a disc running in a parallel format, the device offers the possibility of single-step, multiplexed allergens screening. The device is capable of reporting a result within 30 min. It has many desirable merits including fast and multiplexed analysis, low cost, single-step operation, minimal sample volume, less discomfort and most importantly increased safety as patients are no longer susceptible to possible anaphylactic shock reactions induced by the common skin-prick-test. The flexibility of the flow control within the device makes it suitable to a wide range of CBBs. (C) 2012 Elsevier B.V. All rights reserved.