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

通过透射电子显微镜研究spematozoa的超微结构通常会根据所采用的方法出现解释问题,具体取决于样品是先离心到固定物还是浸入粘性凝胶中。解释的主要问题是: 在成熟过程中引起的囊泡位置的变化以及精浆蛋白在精子膜表面吸附的改变。我们研究的目的是为超微结构研究提供一种治疗精子的原始新方法。我们的方法基于将动物组织用作生物容器,其中包括精索悬浮液。我们使用从成熟的Rasa aragonesa公羊中采集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1周大鸡 (Gallus gallus) 的2.5厘米长的肠段 (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碎片向内放置。其中一个边缘用细细的缝合丝绑着。以最佳实验条件和数量注射精子悬浮液。最后,以与另一段边缘相同的方式用丝将肠段的仍然开放的边缘绑扎。通过使用这种技术,我们可以在超微结构水平上进行适当的形态学研究。此外,正确保留了靶细胞超微结构成分的功能关系。

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