The study of the ultrastructure of spematozoa by means of transmission electron microscopy often presents with problems of interpretation according to the method employed, depending on whether samples are either centrifuged previously to the fixation or immersed in viscous gels. The major problems of interpretation are: changes in the location of vesicles originated during the maturation process and modifications in the adsorption of seminal plasma proteins to the sperm membrane surface. The aim of our study is to communicate an original new method for the treatment of spermatozoa for ultrastructural study. Our method is based on the use of animal tissues as biological containers, inside which the spermatic suspensions are included. We developed this method using fresh sperm samples taken from mature Rasa aragonesa rams. As biological container, we used 2.5-cm long segments of the intestine of 1-week-old chickens (Gallus gallus) (diameter around 4 mm). To avoid any influence of digestive enzymes of the mucosa on the sperm surface, we put each intestine fragment inside out by means of microdissection forceps under bifocal optical microscope and cold light. One of the edges was tied with thin suture silk. The sperm suspension was injected in the optimal experimental condition and amount. Finally, the still open edge of the intestine segment was tied with silk in the same way as the other segment edge. By using this technique, we can perform a suitable morphological study at an ultrastructural level. In addition, the functional relationship of the ultrastructural components of the target cells is correctly preserved.

译文

:根据透射电子显微镜对血吸虫超微结构的研究通常会遇到根据所用方法进行解释的问题,这取决于样品是预先离心至固定液还是浸入粘性凝胶中。解释的主要问题是:在成熟过程中产生的囊泡位置的变化以及精浆蛋白对精子膜表面的吸附作用的改变。我们研究的目的是为了交流用于超微结构研究的治疗精子的原始新方法。我们的方法是基于将动物组织用作生物容器,其中包含了精子悬浮液。我们使用从成熟的Rasa aragonesa公羊中提取的新鲜精子样本开发了这种方法。作为生物容器,我们使用了2.5周长的1周龄鸡(鸡胆)的肠段(直径约4毫米)。为了避免粘膜消化酶对精子表面的任何影响,我们在双焦点光学显微镜和冷光下通过显微解剖钳将每个肠段内翻。边缘之一用细缝线缝合。以最佳实验条件和最佳剂量注射精子悬浮液。最后,肠段的仍然开放的边缘与另一段边缘的相同方式用丝绸绑在一起。通过使用这种技术,我们可以在超微结构水平上进行合适的形态学研究。另外,正确保留了靶细胞超微结构成分的功能关系。

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