Microscale Bioseparations Laboratory and Biomedical Engineering Department, Rochester Institute of Technology, 160 Lomb Memorial Drive, Rochester, New York 14623, United States.
Anal Chem. 2024 Oct 1;96(39):15711-15719. doi: 10.1021/acs.analchem.4c03336. Epub 2024 Sep 18.
Cell viability studies are essential in numerous applications, including drug development, clinical analysis, bioanalytical assessments, food safety, and environmental monitoring. Microfluidic electrokinetic (EK) devices have been proven to be effective platforms to discriminate microorganisms by their viability status. Two decades ago, live and dead (. ) cells were trapped at distinct locations in an insulator-based EK (iEK) device with cylindrical insulating posts. At that time, the discrimination between live and dead cells was attributed to dielectrophoretic effects. This study presents the continuous separation between the live and dead . cells, which was achieved primarily by combining linear and nonlinear electrophoretic effects in an iEK device. First, live and dead . cells were characterized in terms of their electrophoretic migration, and then the properties of both live and dead . cells were input into a mathematical model built using COMSOL software to identify appropriate voltages for performing an iEK separation in a T-cross iEK channel. Subsequently, live and dead cells were successfully separated experimentally in the form of an electropherogram, achieving a separation resolution of 1.87. This study demonstrated that linear and nonlinear electrophoresis phenomena are responsible for the discrimination between live and dead cells under DC electric fields in iEK devices. Continuous electrophoretic assessments, such as the one presented here, can be used to discriminate between distinct types of microorganisms including live and dead cell assessments.
细胞活力研究在许多应用中至关重要,包括药物开发、临床分析、生物分析评估、食品安全和环境监测。微流控电动(EK)设备已被证明是通过其生存状态区分微生物的有效平台。二十年前,活细胞和死细胞在基于绝缘体的 EK(iEK)设备中使用圆柱形绝缘柱被捕获到不同的位置。当时,活细胞和死细胞的区分归因于介电泳效应。本研究提出了在 iEK 设备中通过组合线性和非线性电泳效应来实现活细胞和死细胞的连续分离。首先,对活细胞和死细胞的电泳迁移进行了表征,然后将活细胞和死细胞的特性输入到使用 COMSOL 软件构建的数学模型中,以确定在 T 型交叉 iEK 通道中进行 iEK 分离的适当电压。随后,通过电迁移图的形式成功地在实验中分离了活细胞和死细胞,实现了 1.87 的分离分辨率。本研究表明,在线性和非线性电泳现象的作用下,iEK 设备中的直流电场能够区分活细胞和死细胞。连续的电泳评估,如本文所展示的,可以用于区分不同类型的微生物,包括活细胞和死细胞的评估。