基于微流控芯片的牛乳体细胞均匀分布与计数方法研究  

A study on the uniform distribution and counting method of raw cow's milk somatic cells based on microfluidic chip

作　　者：周围[1] 王明慧 安广鑫 郑泓飙 李星宇 孟庆宜[2] Zhou Wei;Wang Minghui;An Guangxin;Zheng Hongbiao;Li Xingyu;Meng Qingyi(School of Mechanical Engineering,Hebei University of Technology,Tianjin 300401,China;School of Energy Engineering,Tianjin Sino-German University of Applied Sciences,Tianjin 300350,China)

机构地区：[1]河北工业大学机械工程学院,天津300401 [2]天津中德应用技术大学能源工程学院,天津300350

出　　处：《红外与激光工程》2023年第8期245-257,共13页Infrared and Laser Engineering

基　　金：河北省自然科学基金面上项目(B2021202038)。

摘　　要：生鲜牛乳中的体细胞数量是判断奶牛是否患有乳房炎的重要依据。针对牛乳在取样过程中细胞贴壁沉降等原因造成体细胞分布不均匀,从而导致体细胞计数不具有代表性的问题,文中提出了一种基于九宫格型微流控芯片使体细胞分布均匀并提升计数准确率的方法。首先在Comsol仿真的基础上制备了九宫格型微流控芯片,提高了细胞分布的均匀度。其次研制了集染色、搅拌于一体的负压进样系统,保证在进样过程中持续保持体细胞分布的均匀度和不受空气的污染。并配合芯片研制了微型显微成像系统,对芯片的九个观测腔拍摄图像。最后通过图像处理的方法对体细胞进行计数,并判断奶牛乳房的健康状况。实验结果表明,每组九张图像体细胞数量的标准差系数均小于等于1.61%,系统计数准确率可达到99.23%。该研究方法为奶牛乳房炎的检测与预防奠定了基础。Objective The somatic cell count(SCC)in raw milk is an important basis for determining whether a cow is suffering from mastitis.Identifying cows with mastitis by testing the SCC,then isolating and treating them as early as possible,can effectively prevent the spread of bacteria in the herd to reduce consequential economic losses.However,traditional methods may lead to uneven distribution of somatic cells during milk sampling,such as cell adhesion settlement,and unrepresentative somatic cell count due to lack of matching imaging system.In this paper,a method is proposed which is based on the nine-cell grid microfluidic chip to make somatic cell evenly distributed and develop a two degree of freedom displacement platform equipped with a micro lens to improve the counting accuracy.Methods Firstly,a simulation was performed to verify the uniformity of the somatic cell distribution within the chip observation cavities(Fig.1).And based on the simulation results,a nine palace grid microfluidic chip was prepared(Fig.2).Secondly,a two-degree-of-freedom displacement platform(Fig.6)equipped with a micro-camera lens is developed,which can automatically take images of the nine observation cavities of the chip,making image acquisition more convenient.Finally,somatic cells were counted by image processing(Fig.3),so as to verify the uniformity of somatic cell distribution,obtain the counting accuracy,and judge the health status of cow udder.Results and Discussions 20 cows were randomly selected from local pastures to verify the performance of the proposed method.From the data in Tab.1,it can be seen that the standard deviation coefficient of the SCC in each group of nine images is less than or equal to 1.61%,which verifies the uniformity of the distribution of somatic cells in the nine observation cavities of the microfluidic chip.The system ensures the uniform distribution of somatic cells and renders the taken samples more representative.As can be seen from Fig.9(b),the maximum absolute value of the relative error of the syste

关 键 词：奶牛乳房炎 微流控芯片 微型显微镜系统 图像处理 细胞计数 标准差系数 

分 类 号：TH89[机械工程—仪器科学与技术]