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A Nut-and-Bolt Microfluidic Mixing System for the Rapid Labeling of Immune Cells with Antibodies

1
Department of Integrative Biomedical Science and Engineering, Graduate School, Kookmin University, 77 Jeongneung-ro, Seongbuk-gu, Seoul 02707, Korea
2
School of Mechanical Engineering and Department of Integrative Biomedical Science and Engineering, Graduate School, Kookmin University, 77 Jeongneung-ro, Seongbuk-gu, Seoul 02707, Korea
*
Author to whom correspondence should be addressed.
Micromachines 2020, 11(3), 280; https://doi.org/10.3390/mi11030280
Received: 22 January 2020 / Revised: 1 March 2020 / Accepted: 8 March 2020 / Published: 9 March 2020
(This article belongs to the Special Issue Microfluidics-Based Point-of-Care Diagnostics)
A nut-and-bolt microfluidic system was previously developed for a point-of-care (POC) human immunodeficiency virus (HIV) test and was able to acquire images of CD4 (cluster of differentiation 4) + T-lymphocytes in a sample drop of blood followed by image analysis. However, as the system was not fully integrated with a sample reaction module, the mixing of the sample with the antibody reagent was carried out manually. To achieve a rapid reaction with a reduced amount of costly reagent in a POC diagnostic system, an efficient sample mixing function must be implemented. Here, we propose a novel method to drastically accelerate the process of sample mixing and increase the reaction rate in the nut-and-bolt microfluidic system, where the sample is mixed with the reagent in a reaction chamber formed by connecting a nut with a bolt-like sample cartridge. The mixing is facilitated by rotating the sample cartridge bidirectionally using a DC motor, which agitates the sample in a chaotic manner. A microbead complex formed by the avidin–biotin interaction was used as a model reaction system to examine the feasibility of our mixing module. We found that the reaction time for the avidin–biotin binding by mixing was 7.5 times shorter than in the incubation method, achieving a reaction efficiency of over 95%. The performance of our mixing system was further demonstrated by measuring the concentration of CD4 cells labeled with a fluorescent antibody in the blood sample. The antigen–antibody reaction mixing was faster by a factor of 20, reaching a reaction efficiency comparable to the conventional incubation method. View Full-Text
Keywords: nut-and-bolt microfluidics; reaction efficiency; mixing; antibody labeling; CD4+ T-cells; fluorescence imaging nut-and-bolt microfluidics; reaction efficiency; mixing; antibody labeling; CD4+ T-cells; fluorescence imaging
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Imran, J.H.; Kim, J.K. A Nut-and-Bolt Microfluidic Mixing System for the Rapid Labeling of Immune Cells with Antibodies. Micromachines 2020, 11, 280.

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