基于免疫功能异常性不育症检测的新型微悬臂梁阵列传感器的关键技术及机理研究

Immune dysfunction is a clinically infertility and difficult to cure. Anti-sperm antibodies (AsAb) detection for this disease is common clinical tool. However, a common detection method can't reflect real time binding process between AsAb and sperm antigens, and the activity of sperm in real time. In previous studies, we have set up a microcantilever sensor system and development immobilization antibody on the gold surface of a microcantilever. As microcantilever technology have advantages real-time, high sensitivity, label-free and other advantages, we investigate binding studies of anti-sperm antibodies and antigens using this technique. Firstly, a deflection detection system for microcantilever array is setup based on mirror translational mode scanning. A controllable humidity and temperatures environment is built for the reaction process in suitable condition. Antibody molecules are modified on the gold surface of microcantilever in micro pipeline. A micro-droplet is injected by micro-flow module for micro reactor for banding reaction between antigen and antibody on the gold surface of microcantilever. The banding site on the sperm for antibody bound to the antigen is characterized by fluorescence -labeled and sperm activity and anti-sperm antibody is evaluated by enzyme-linked immune sorbent assay.The subject can be studied on activity of different sperm, the banding site on the sperm for antibody and sperm antigens interfacial stress on the mode of transmission on the surface of microcantilever. A new method of detection immune dysfunction infertility will provides in this work.

免疫功能异常是一种临床难治愈的不孕症。抗精子抗体（AsAb）检测是该病情确诊的常用临床手段。常见检测手段不能实时反映AsAb绑定过程和不同精子的活性。在前期研究中，我们开展了微梁系统搭建和微悬臂梁免疫传感的研究。基于实时、高灵敏度和非标定等优势，本课题拟搭建新型微悬臂梁阵列传感系统进行AsAb和精子的免疫研究。首先采用基于平面镜平动扫描方式实现微梁阵列偏转照射；设计一种可控温和控湿反应环境，使反应在适宜的条件下进行；通过微流动技术在微梁表面修饰抗体分子，利用微流动实现抗原微液滴滴到微梁上修饰抗体的表面，实现抗原抗体的微反应，利用微梁弯曲实时检测免疫反应过程。利用荧光标记抗体研究参与绑定的抗原在精子上具体位置，利用酶联免疫吸附法对精子活性和抗精子抗体进行表征。本课题可以研究不同精子活性及精子上精子抗原存在位置及其和抗体作用时对界面应力的传播方式，为免疫功能异常的不孕症提供一个新型研究手段。

不孕症检测是解决全世界低出生率和老龄化人口的重要挑战。免疫功能异常是一种临床难治愈的不孕症。精液分析是诊断评估男性对不孕症贡献的基石。在本项目研究中，首先搭建了一种新型基于平面镜平动扫描方式实现微梁阵列偏转扫描的系统。其次并实现高精度控温反应环境下，对精子在纳米量级上的运动导致微梁的涨落信号进行实时检测。研究中发现微梁涨落的幅度大小与精子活力的大小相关，通过计算微梁涨落曲线的方差来量化精子活力。利用微梁传感器研究了精子在不同药物（酒精与伟哥）作用下的活力变化，实现对增强或抑制精子活力的药物筛选。最后研究了环境温度对精子活力的影响。实验表明，精子活动导致不同的在微梁的不同程度的涨落，而对微梁这种信号的分析可以得到精子活性的数据。这项技术为精子的特性研究提供了一个全新的视角。通过结合传统的测量方法，生殖医学医生和研究人员可以在不同的角度对精子活性特性进行研究。.此外在本项目资助下，还利用微悬臂梁传感技术对偶氮苯顺反异构化过程和橡胶弹性粒子溶胀特性等进行研究，并进行了废食用油热解产物作为一种有潜力的浮选捕收剂研究。