一种新型无标记白细胞计数检测系统的设计

A novel design of label-free white blood cell count detection system

目的人体白细胞(WBC)总数是重要的生理指标,是临床检验的核心依据。针对基层医疗和灾害应急救援伤(病)等条件下对设备便携和操作简便的要求,提出一种基于微流控芯片的"无标记"WBC总数检测方法。方法以微流控芯片为检测载体,芯片内预封装溶血剂,将约10μl新鲜血液注入芯片微腔中进行溶血处理;针对样本特性设计了细胞暗视场散射显微成像光学系统,对于无染色标记的WBC可获得高质量对比度图像;基于边界跟踪算法和分水岭算法原理优化了图像目标识别算法,实现黏连细胞分割和准确计数。结果实际血液样本测试表明,该系统与Sysmex XE-5000血液分析仪的结果比例差异为0. 986,P> 0. 05,表明两组测量具有一致性。在WBC浓度≥3. 0×109/L的重复性测试结果中,CV值<4. 0%,曲线拟合R2=0. 9996。结论该系统对WBC计数的主要技术指标均满足临床检验使用要求。

Objective The white blood cell( WBC) count is an important physiological index of the human body. In order to meet the requirements of portable and easy operation of equipment during basic medical treatment and disaster emergency rescue,this paper proposed a " label-free" WBC count detection method based on microfluidic chips. Methods The microfluidic chip was used as the detection supporter and the hemolytic agent was packaged in the chip. Then,about10 μl fresh blood was injected into the microcavity of the chip for hemolysis. A forward scattering optical cell imaging optical system was designed according to the sample characteristics so that high quality contrast images could be obtained for unlabeled WBCs. Based on the boundary tracking algorithm and watershed algorithm,the algorithm of image target recognition was optimized to achieve segmentation and accurate counting of the adhered cells. Results The proportional differences between the test results of our system and those of the Sysmex XE-5000 blood analyzer was 0. 986 with P >0. 05,indicating consistency between the two groups. The CV was less than 4. 0% in the repeatability test result of the WBC concentration of 3. 0 × 109/L or above,and the value of curve fitting R2 was equal to 0. 9996. Conclusion The main technical indicators of this system for WBC counting all meet the requirements for clinical testing.