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通过光学显微镜估计的细胞特征。目前将精子计数、形态学和活力确定为主要终点。虽然这些参数提供了关于睾丸中配子产生的有价值的读数，但精子的功能特征，特别是它们的受精能力还没有得到充分的解决。近年来，一些生化测试被开发出来，以更好地识别精子的功能能力。这些策略对于改善人类和兽医学中的精子分析和精子选择是强制性的。我们的团队已经进入了另一条轨道，探索应用新颖和创新的光学策略进行精子分析。我们在许多物种中获得了许多关于精子完整性和功能的新见解，并创造了几种新的诊断策略。我们假设，非侵入性的光学策略可以应用于获得个人精子的功能参数。这些终点将使我们能够更精确地描述精子在受精发生之前需要经过的一连串事件。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，我们描述了各个精子的各种参数，为精子质量的完整性和适应症提供了见解。我们认为，如多参数流式细胞术（MFC）结合细胞分选和拉曼显微光谱（RM）技术提供了出色的诊断机会。我们期望，这些发现改变了对精子受精能力机制的理解。几个终点的组合提供了评估精子质量/功能的可能性，远远超出了细胞学/显微镜参数。新的光学技术免除了技术限制，现在可以探索单个精子。我们可以定量地鉴定没有缺陷、有一种缺陷或有几种缺陷的精子亚群。这是第一次，我们可以同时确定各种功能细胞特征之间的关系。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选的新应用。

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.