Department of Biomedical Engineering , Kyung Hee University , Yongin-si , Gyeonggi-do 17104 , Republic of Korea.
Department of Biomaterials Science, Life and Industry Convergence Institute , Pusan National University , 1268-50 Samrangjin-ro , Miryang 50463 , Republic of Korea.
Anal Chem. 2018 Jul 3;90(13):8254-8260. doi: 10.1021/acs.analchem.8b01804. Epub 2018 Jun 19.
Major challenges of miniaturizing flow cytometry include obviating the need for bulky, expensive, and complex pump-based fluidic and laser-based optical systems while retaining the ability to detect target cells based on their unique surface receptors. We addressed these critical challenges by (i) using a viscous liquid additive to control flow rate passively, without external pumping equipment, and (ii) adopting an immunobead assay that can be quantified with a portable fluorescence cell counter based on a blue light-emitting diode. Such novel features enable pumpless microflow cytometry (pFC) analysis by simply dropping a sample solution onto the inlet reservoir of a disposable cell-counting chamber. With our pFC platform, we achieved reliable cell counting over a dynamic range of 9-298 cells/μL. We demonstrated the practical utility of the platform by identifying a type of cancer cell based on CD326, the epithelial cell adhesion molecule. This portable microflow cytometry platform can be applied generally to a range of cell types using immunobeads labeled with specific antibodies, thus making it valuable for cell-based and point-of-care diagnostics.
微流控流式细胞术面临的主要挑战包括避免使用庞大、昂贵且复杂的基于泵的流体系统和基于激光的光学系统，同时保留基于其独特表面受体检测靶细胞的能力。我们通过以下两种方法解决了这些关键挑战：（i）使用粘性液体添加剂来被动控制流速，而无需外部泵送设备，以及（ii）采用免疫珠测定法，该方法可以使用基于蓝光发光二极管的便携式荧光细胞计数器进行定量。这些新颖的特征使得无需泵的微流控流式细胞术（pFC）分析可以通过将样品溶液简单地滴加到一次性细胞计数室的入口储液器中即可实现。通过我们的 pFC 平台，我们实现了在 9-298 个细胞/μL 的动态范围内可靠的细胞计数。我们通过基于上皮细胞黏附分子 CD326 来识别一种癌细胞，证明了该平台的实际应用价值。这种便携式微流控流式细胞术平台可以使用标记有特定抗体的免疫珠来应用于各种类型的细胞，因此非常适合基于细胞的即时诊断。
