Biochemical and physiological characterization of human spermatozoa using novel optical technologies - A new understanding of sperm functional competence

临床和实验精子分析依赖于光学显微镜估计的细胞特征。目前，精子数量、形态和活动率被确定为主要终点。虽然这些参数提供了关于睾丸配子产生的有价值的读数，但精子的功能特征，特别是它们受精的能力还没有得到充分的研究。近年来，为了更好地鉴定精子的功能能力，开发了几种生化测试方法。这些策略对于改善人类和兽医中的精子分析和精子选择是强制性的。我们的团队通过探索应用新颖和创新的光学策略进行精子分析，进入了一条替代的轨道。我们对许多物种的精子完整性和功能有了许多新的见解，并创造了几种新的诊断策略。我们假设，非侵入性光学策略可以应用于获取单个人类精子的功能参数。这样的终点将使我们能够更准确地描述精子在受精之前需要通过的一系列事件。这些新发现将改变我们对精子功能的概念，并可能为精子评估提供新的策略。在这个项目中，我们描述了个体精子的各种参数，为了解精子的完整性和精子质量的指标提供了见解。我们认为，多参数流式细胞术(MFC)与细胞分选和拉曼显微光谱分析(RM)相结合的技术提供了绝佳的诊断机会。我们预计，这些发现将改变人们对精子受精机制的理解。几个终点的组合提供了评估精子质量/功能的潜力，远远超出了细胞学/显微参数。新的光学技术免除了技术限制，现在可以探索个人精子。我们可以定量地鉴定出没有、有一个或几个缺陷的精子亚群。第一次，我们可以同时确定各种功能细胞功能之间的关系。我们的假设驱动的方法有可能彻底改变我们对精子功能的理解，并可能导致精子分析和分类的新应用。

Clincial and experimental sperm analysis relies on cellular features estimated by light microscopy. Sperm count, morphology and motility are currently determined as the primary endpoints. While these parameters present valuable readouts on gamete production in the testis, the functional features of spermatozoa and specifically their competence to fertilize is not adequately addressed. In recent years several biochemical tests were developed to better identify sperm functional competence. Such strategies are mandatory to improve sperm analysis and sperm selection in human and veterinary medicine. Our group has entered an alternative track by exploring the application of novel and innovative optical strategies for sperm analysis. We obtained many new insights into sperm integrity and function in a number of species and created several new diagnostic strategies. We hypothesize, that non-invasive optical strategies can be applied to obtain functional parameters in individual human spermatozoa. Such endpoints will enable us to describe more precisely the cascade of events a sperm needs to pass before fertilization can occur. Such new findings will change our concept for sperm function and may provide novel strategies for sperm assessment. In this project we describe various parameters on individual sperm providing insights into the integrity and indications for sperm quality. We consider that techniques such as Multiparametric Flow Cytometry (MFC) coupled to cell sorting and Raman Micro-spectroscopy (RM) offer outstanding diagnostic opportunities. We expect, that such findings change the understanding of the mechanisms rendering spermatozoa fertilization competent. Combination of several endpoints offers the potential to assess sperm quality/function far beyond cytological/microscopic parameters. The new optical technologies waived technical limitations and now enable exploration of individual sperm. We can identify quantitatively subpopulations of sperm with none, one or several defects. For the first time, we can determine simultaneously the relationship between various functional cellular features. Our hypothesis-driven approach has the potential to revolutionize our understanding of sperm function and may lead to novel applications for sperm analysis and sorting.