Kwok Ho Chin, Lau Pui Man, Wu Shu Yuen, Ho Ho Pui, Gao Minghui, Kwan Yiu Wa, Wong Chun Kwok, Kong Siu Kai
Department of Electronic Engineering, Center for Advanced Research in Photonics, The Chinese University of Hong Kong, Shatin, Hong Kong, China.
Programme of Biochemistry, School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong, China.
Micromachines (Basel). 2016 Feb 27;7(3):38. doi: 10.3390/mi7030038.
A lab-on-a-disc (LOAD) is a centrifugal microfluidic set-up based on centrifugal force without using micro-pumps to drive reagents and cells to various chambers through channels and valves for reactions. A LOAD coated with conductive transparent indium tin oxide (ITO) for thermal control was developed to screen allergy-blocking agents. When the acridine orange (AO)-loaded KU-812 human basophilic cells were activated in the LOAD by stimuli, AO trapped in the cytoplasmic granules was released externally as an allergic mediator mimetic to report degranulation. This response was monitored by fluorescence when the released AO in supernatant had been transferred, with a higher spinning speed, from the reaction chamber to detection chamber in the LOAD where AO reacted with exogenous DNA. We report here the principles of the system and an improved LOAD set-up with the ITO-coated glass resistive microheater to run assays at 37 °C. By using this platform, we demonstrate here for the first time that triptolide, an active ingredient from the Chinese medicine herb Hook f. was able to suppress the fMLP-mediated degranulation in basophils. This serves as an example how LOADs can be used to screen agents to alleviate symptoms of allergy.
盘式实验室(LOAD)是一种基于离心力的离心微流控装置,无需使用微型泵来驱动试剂和细胞通过通道和阀门进入各个腔室进行反应。为了筛选抗过敏剂,开发了一种涂覆有用于热控制的导电透明氧化铟锡(ITO)的LOAD。当负载吖啶橙(AO)的KU-812人嗜碱性细胞在LOAD中被刺激激活时,被困在细胞质颗粒中的AO作为模拟过敏介质释放到外部,以报告脱颗粒情况。当上清液中释放的AO以更高的旋转速度从反应腔转移到LOAD中的检测腔时,通过荧光监测这种反应,在检测腔中AO与外源DNA发生反应。我们在此报告该系统的原理以及一种改进的LOAD装置,该装置带有涂覆ITO的玻璃电阻微加热器,可在37°C下进行检测。通过使用这个平台,我们首次证明了中药雷公藤中的活性成分雷公藤内酯醇能够抑制嗜碱性粒细胞中fMLP介导的脱颗粒。这为例示了LOAD可如何用于筛选减轻过敏症状的药物。