【作者】 刘珊；

【导师】 周传清；

【作者基本信息】 上海交通大学 ， 生物医学工程（专业学位）， 2020， 硕士

【摘要】 哺乳动物的输卵管是配子运输、受精和着床前胚胎发育的重要环境,其中输卵管内的纤毛运动在生殖过程中发挥着重要作用,比如将受精卵运送至子宫进行着床。尽管越来越多的证据表明不孕不育与输卵管纤毛受损有着密切的关系,但是人们对于输卵管在纤毛正常情况和受损情况下其生殖功能的机理尚不完全清楚。输卵管纤毛摆动频率(Cilia Beat Frequency,CBF)是反映输卵管纤毛上皮细胞运动的重要的参数。大鼠可作为人类生殖研究的重要模型,在目前我们对于大鼠输卵管内纤毛摆动频率的检测方法中,如传统的光学显微镜和共聚焦显微镜,由于这两种技术的成像深度有限,需要将输卵管从动物体内分离出来,将其打开后纤毛暴露于腔体的表面,这个过程可能会影响甚至改变纤毛的生物力学环境,从而可能导致对生理过程解释的不准确。然而,光学相干层析成像技术(Optical Coherence Tomography,OCT)一种无创的、非侵入式对生物组织进行三维高分辨率成像的技术,能够克服传统显微成像的不足。因而,本文针对传统检测技术的不足之处,基于OCT的大鼠输卵管成像技术,获得大鼠在体输卵管结构与功能成像,为以后临床中输卵管疾病的诊断和治疗提供更多的信息。具体研究内容和创新点包括:首先,完善及优化基于OCT系统的系统和算法,获取生物组织一系列二维横断面图像。因为在OCT系统的图像采集过程中容易混入噪声,为提高成像质量,本文通过图像配准和中值滤波的方法对OCT图像进行处理,抑制噪声,提高计算CBF的准确性。其次,由于大鼠气管组织的易操作和易获取的优点,本文通过对比和分析传统数字视频显微镜成像技术和OCT成像技术,分别计算计算离体大鼠气管组织处纤毛的摆动频率,验证了基于OCT成像技术数算法的有效性,为后续分析输卵管纤毛活动提供可靠的技术手段。最后,以在体大鼠输卵管为研究对象,获取在体大鼠输卵管的三维结构信息,定量分析大鼠输卵管纤毛摆动频率的时空特征,获得输卵管纤毛位置的定位图像。统计输卵管壶腹部纤毛的CBF,揭示该部位纤毛的CBF有一个较大的频率范围,表明纤毛摆动的不均匀性。随机选取壶腹部的4个局部褶皱作为感兴趣区域,经Wilcoxon秩和检验的统计分析,表明在体大鼠输卵管壶腹部内不同褶皱区域纤毛的CBF并无明显差异,说明对于同一大鼠输卵管壶腹部管腔内不同褶皱区域纤毛摆动频率范围具有一致性。本文通过对大鼠输卵管纤毛活动的研究将有助于进一步揭示纤毛活动异常对于输卵管功能的影响的机理,拓展了OCT成像在生殖领域的应用,能够为临床检测和治疗输卵管结构和功能异常的疾病提供新的思路和方法。更多还原

【Abstract】 The oviduct(or fallopian tube)plays a key role in gametes transport,fertilization and preimplantation embryo development with cilia beat at a native frequency in mammals.The ciliary movement in the mammalian fallopian tubes also plays an important role in reproduction,such as transporting fertilized oocytes into the uterus for implantation.Despite the increasing evidence suggests that the infertility has close relationship with the cilia damaged of the mammal oviduct,the ciliary motion of the mechanism in both normal conditions and impaired reproductive process is not fully clear.The mouse is an important model in the research of human reproduction.However,the traditional bright field microscopy and confocal microscope have been used to map the cilia and measure the cilia beat frequency(CBF).However,the oviduct must be dissected from the body,and opened up to expose the surface of the ciliated lumen for imaging.This might change the biomechanical environment of the cilia,which may lead to inaccurate interpretation of physiological processes.Optical coherence tomography(OCT)is a noninvasive high-resolution imaging technique capable of capturing three-dimension micrometer scale cross-sectional images of highly scattering biological tissue.Therefore,we focused on the3 D imaging approach by our custom-designed spectral-domain OCT for the vivo mouse oviduct so as to address the limitation of traditional detection technology in this paper,which can provide more information for the diagnosis and treatment of fallopian tube diseases in the future.The research work can be summarized as follows:Firstly,the algorithm and system of the oviduct OCT imaging is improved and optimized.It is easy to introduce the noises into the OCT signals during the further acquisition.In order to improve the image quality,the OCT image is processed by image registration and median filtering to suppress noise and improve the accuracy of CBF calculation.Secondly,to validate measurements made by our OCT,comparisons were made using ex vivo mouse airway surface imaging between OCT and digital microscopy respectively.The result can provide a theoretical basis for subsequent analysis of the ciliary motion.In vivo,we obtained the three-dimensional structure information of the mouse oviduct,measured the CBF with the OCT system and map the cilia location.Next,we further analyzed the ciliary motion through choosing areas of interest from the mapping of the CBF location.The CBF in multiple regions on the same oviductal fold were compared by the method of the twosided Wilcoxon rank-sum tests.The result suggests that cilia were able to beat at distinct frequencies,which was also observed in smaller folded regions on the same fold.The cilia beat has a similar range of frequencies within a local region in the same mouse oviduct fold.The approach will benefit the study in vivo mammalian reproduction so as to explore the link between infertility and activities.In summary,the study on the ciliary motion of the mouse oviduct will help to further reveal the mechanism of the influence of abnormal ciliary activity on the infertility,and expand the application of the OCT imaging in mammalian reproduction,which might provide a new idea and method for the disease diagnose and treatment about abnormal structure and function of the fallopian tube.更多还原