摘要
基于可视化血小板超微结构的血液活检技术在急性缺血性脑卒中等疾病的监测与诊断领域有重要应用。但目前血小板的免疫荧光染色严重依赖手动操作,任务繁重、效率低且一致性差,难以应对日益增长的血液样本监测需求。为了解决人工染色的不足以及弥补国内全自动血小板免疫荧光染色设备的空白,首创了一种血小板全自动免疫荧光染色控制系统,其主要包括电源模块、主控模块、传感反馈模块、运动执行模块和上位机软件等。该控制系统能够实现十样本通量的免疫荧光染色操作的精准控制,操作步骤覆盖打孔、封闭、一抗标记、一抗漂洗、二抗标记、二抗漂洗、PFA(polyformaldehyde fixative,多聚甲醛固定液)固定、漂洗等。测试结果表明,所设计的控制系统可将X、Y、Z运动轴的重复定位精度控制在±0.1 mm内,滴加微量试剂精度达到1μL,滴加大量试剂精度达到1 mL,多次实验的故障率小于1%,这可在降低血小板免疫荧光染色成本的同时获得清晰度、可判度与人工染色相当的染色结果。血小板全自动免疫荧光染色控制系统具有稳定性高、准确性高及效率高的优点,所获得的血小板超微结构荧光染色结果在超分辨率显微镜下的清晰度很高,这可为基于超分辨技术的血小板超微结构检测提供高效的实验工具,具有重要的工程意义。
Blood biopsy technology based on visualized platelet ultrastructure has important applications in the monitoring and diagnosis field of acute ischemic stroke and other diseases.However,current platelet immunofluorescence staining heavily relies on manual operation,with heavy workload,low efficiency and poor consistency,and it is difficult to cope with the increasing demand for blood sample monitoring.In order to solve the shortage of manual staining and make up for the blank of automatic platelet immunofluorescence staining equipment in China,an automatic platelet immunofluorescence staining control system was invented,which mainly included power module,main control module,sensor feedback module,motion execution module and upper computer software.The control system could realize the accurate control of the immunofluorescence staining operations with ten sample fluxes,including drilling,sealing,primary antibody labeling,primary antibody rinsing,secondary antibody labeling,secondary antibody rinsing,PFA(polyformaldehyde fixative)fixation and rinsing.The test results showed that the designed control system could control the repeated positioning accuracy of the X,Y and Z axes within±0.1 mm,the accuracy of adding trace reagents reached 1μL,the accuracy of adding a large amount of reagents reached 1 mL,and the failure rate of multiple experiments was less than 1%.It could reduce the cost of platelet immunofluorescence staining while obtaining a clear and determinable staining result comparable to manual staining.The automatic platelet immunofluorescence staining control system has the advantages of high stability,high accuracy and high efficiency,and the obtained fluorescence staining results of platelet ultrastructure have high clarity under super-resolution microscope,which can provide an efficient experimental tool for the detection of platelet ultrastructure based on super-resolution technology,and has important engineering significance.
作者
周文韬
樊帆
林成旭
廖广兰
刘智勇
ZHOU Wentao;FAN Fan;LIN Chengxu;LIAO Guanglan;LIU Zhiyong(School of Mechanical Science and Engineering,Huazhong University of Science and Technology,Wuhan 430074,China)
出处
《工程设计学报》
CSCD
北大核心
2024年第5期681-688,共8页
Chinese Journal of Engineering Design
关键词
血小板超微结构检测
全自动免疫荧光染色
控制系统
platelet ultrastructure detection
automatic immunofluorescence staining
control system
