Kasai Shigenobu, Prasad Ankush, Kumagai Ryoma, Takanohashi Keita
Graduate Department of Electronics, Tohoku Institute of Technology, Sendai 982-8577, Japan.
Department of Biophysics, Faculty of Science, Palacký University, 783 71 Olomouc, Czech Republic.
Biology (Basel). 2022 Apr 1;11(4):549. doi: 10.3390/biology11040549.
The method to diagnose mastitis is generally the somatic cell count (SCC) by flow cytometry measurement. When the number of somatic cells in raw milk is 2.0 × 10 cells/mL or more, the condition is referred to as mastitis. In the current study, we created a milk cell chip that serves as an electrochemical method that can be easily produced and used utilizing scanning electrochemical microscopy (SECM). The microelectrode present in the cell chip scans, and the difference between the oxygen concentration near the milk cell chip and in bulk is measured as the oxygen (O) reduction current. We estimated the relationship between respiratory activity and the number of somatic cells in raw milk as a calibration curve, using scanning electrochemical microscopy-somatic cell count (SECM-SCC). As a result, a clear correlation was shown in the range of 10 cells/mL to 10 cells/mL. The respiration rate (F) was estimated to be about 10-16 mol/s per somatic cell. We also followed the increase in oxygen consumption during the respiratory burst using differentiation inducer phorbol 12-myristate 13-acetate (PMA) as an early stage of mastitis, accompanied with an increase in immune cells, which showed similar results. In addition, we were able to discriminate between cattle with mastitis and without mastitis.
乳腺炎的诊断方法通常是通过流式细胞术测量体细胞计数（SCC）。当原料乳中的体细胞数为2.0×10⁵个细胞/毫升或更多时，这种情况被称为乳腺炎。在当前的研究中，我们创建了一种乳细胞芯片，它作为一种电化学方法，利用扫描电化学显微镜（SECM）可以很容易地生产和使用。细胞芯片中的微电极进行扫描，并测量乳细胞芯片附近和本体中的氧浓度差异作为氧（O）还原电流。我们使用扫描电化学显微镜-体细胞计数（SECM-SCC）作为校准曲线，估计了呼吸活性与原料乳中体细胞数之间的关系。结果，在10⁵个细胞/毫升至10⁷个细胞/毫升的范围内显示出明显的相关性。估计每个体细胞的呼吸速率（F）约为10⁻¹⁶摩尔/秒。我们还使用分化诱导剂佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）作为乳腺炎的早期阶段，跟踪呼吸爆发期间氧气消耗的增加，同时免疫细胞也增加，结果相似。此外，我们能够区分患有乳腺炎和未患乳腺炎的奶牛。
