Spectromicroscopy for biochemical analysis of sperm

描述（申请人提供）：我们建议开发 X射线光谱显微镜，使其能够应用于生物化学成像， 正常和异常精子。这将通过聚焦“软”X射线（在此 情况下，光子能量为200-800 eV的X射线）到最小远场 任何波长的电磁辐射的焦点，并扫描干燥或 冷冻的水合样本通过该焦点以形成图像。图片A 一系列的光子能量将被结合， 化学状态敏感的X射线吸收光谱从整个精子在 空间分辨率优于50 nm。从这些数据来看，主要的生物化学 将通过拟合参考值来确定单个精子中的成分 分离化合物的光谱;此外，预期和可能非预期 将使用principal来研究组件之间的空间相关性 成分分析法这项工作将使用X射线进行 我们在斯托尼布鲁克的研究小组研制的显微镜， 布鲁克海文国家同步加速器光源的波荡器光束线 国家实验室。显微镜将通过增加更好的 顺序排序光学定量光谱，并通过装备低温 具有更精确的压电载物台和更高效率的检测器的显微镜 用于研究冷冻水合标本，而不会造成过度辐射损伤。 这些仪器的发展将遵循我们的目标，即研究 精子形态和生化变化之间的相关性， 更好地了解男性不育的原因。通过获得X射线 在不育男性不同精子形态的光谱显微镜数据，我们 希望能对体外受精（IVP）临床医生在精子选择上起到指导作用 用于卵胞浆内单精子注射（ICSI），以提高成功率 程序的速度。制作了光谱显微数据分析软件 其他研究人员可以免费下载。

Description (provided by applicant): We propose to develop the capabilities of x-ray spectromicroscopy to allow it to be applied to biochemical imaging of normal and abnormal sperm. This will be done by focusing "soft" X rays (in this case, x rays with a photon energy of 200-800 eV) to the smallest far-field focus of electromagnetic radiation of any wavelength, and scanning a dry or frozen hydrated specimen through that focal spot to form an image. Images at a series of photon energies will then be combined to deliver chemical-state-sensitive x-ray absorption spectra from an entire sperm at better than 50 nm spatial resolution. From this data, major biochemical constituents in individual sperm will be determined by fitting to reference spectra of isolated compounds; furthermore, expected and possibly unexpected spatial correlations between components will be studied using principal component analysis methods. This work will be carried out using x-ray microscopes developed by our group at Stony Brook which operate at a soft x-ray undulator beamline at the National Synchrotron Light Source at Brookhaven National Laboratory. The microscopes will be improved by the addition of better order sorting optics for quatitative spectroscopy, and by equipping a cryo microscope with a more accurate piezo stage and a higher efficiency detector for studies of frozen hydrated specimens without excessive radiation damage. These instrumentation developments will be guided by our goal of studying the correlation between morphological and biochemical variations in sperm, in order to better understand the causes of male infertility. By obtaining x-ray spectromicroscopic data on different sperm morphologies in infertile males, we hope to guide the in vitro fertilization (IVP) clinician in the choice of sperm use for intracytoplasmic sperm injection (ICSI) so as to improve the success rate of the procedure. Spectromicroscopic data anaysis software will be made available for free downloading by other researchers as it is developed.