Department of Chemistry, Saint Louis University, 3501 Laclede Ave St. Louis, MO, USA, 63103.
Department of Biomedical Engineering, Institute for Quantitative Health Science & Engineering, Michigan State University, USA.
Anal Methods. 2022 Aug 25;14(33):3171-3179. doi: 10.1039/d2ay00931e.
In this paper, a 3D-printed multi-modal device was designed and fabricated to simultaneously detect nitric oxide (NO) and adenosine triphosphate (ATP) in red blood cell suspensions prepared from whole blood. Once a sample was injected into the device, NO was first detected ( amperometry) using a three-electrode, dual-opposed, electrode configuration with a platinum-black/Nafion coated gold working electrode. After in-line amperometric detection of NO, ATP was detected a chemiluminescence reaction, with a luciferin/luciferase solution continuously pumped into an integrated mixing T and the resulting light being measured with a PMT underneath the channel. The device was optimized for mixing/reaction conditions, limits of detection (40 nM for NO and 30 nM for ATP), and sensitivity. This device was used to determine the basal (normoxic) levels of NO and ATP in red blood cells, as well as an increase in concentration of both analytes under hypoxic conditions. Finally, the effect of storing red blood cells in a commonly used storage solution was also investigated by monitoring the production of NO and ATP over a three-week storage time.
本文设计并制造了一种 3D 打印的多模态设备，用于同时检测全血制备的红细胞悬浮液中的一氧化氮 (NO) 和三磷酸腺苷 (ATP)。一旦将样品注入设备，首先使用具有铂黑/Nafion 涂层金工作电极的三电极、双对电极构型进行电流检测法 (amperometry) 检测 NO。在对 NO 进行在线电流检测后，通过连续泵入整合混合 T 的荧光素/荧光素酶溶液来检测 ATP，并通过通道下方的 PMT 测量产生的光。该设备针对混合/反应条件、检测限 (NO 为 40 nM，ATP 为 30 nM) 和灵敏度进行了优化。该设备用于测定红细胞中基础 (常氧) 水平的 NO 和 ATP，以及在低氧条件下两种分析物浓度的增加。最后，还通过监测在常用储存溶液中储存红细胞时的 NO 和 ATP 产生情况，研究了储存红细胞的影响，储存时间为三周。
