Visualization Study Of Spermatogonia Computer Model And Its Clinical Application

PartⅠSeparation and purification of spermatogonia in ratsObjective In order to study the mechanism of spermatogenesis and its influence factors, the spermatogonia should be separated and purified to provide technological groundwork and cell sources.Methods Continuous enzymatic digestion was used to prepare spermatogenic cell suspension of 9-day-old rats postnatal, and Percoll density gradient centrifugation combined with plating culture method was used to isolate and purify spermatogonia. Results Spermatogonia were mainly distributed in Percoll gradient between 25%-35%. These cells are spherical with large nuclei containing single or multiple nucleoli and small flakes of associated heterochromatin. The Golgi apparatus is distinct, and cytoplasmic organelles are concentrated in a perinuclear region. Conclusion Percoll density gradient centrifugation combined with continuous enzymatic digestion and plating culture method could effectively isolate and purify spermatogonia from 9-day-old rats postnatal. PartⅡComputer-assisted 3D reconstruction of rat testis from serial ultra-thin slice imagesObjective To obtain three-dimensional reconstruction of rat testis from serial sections on the PC-Windows platform.Methods Serial slice images of rat testis were obtained by a digital camera, which were inputted to the computer. After image preprocess in Adobe Photoshop CS, we aligned the serial images and reconstructed the 3D image in Amira 3.1(TGS).Results Internal and external structures of rat testis can be observed from any angles. Conclusion Within the help of Amira 3.1(TGS), 3D reconstructions of tissue can be performed in PC-Windows platform of a common PC, which is easy for popularizing.PartⅢThree-dimensional reconstruction of rat spermatogonia from ultra-thin serial slicesObjective To obtain three-dimensional reconstruction views of rat spermatogonia from ultra-thin serial sections on the PC-Windows platform.Methods Spermatogonia were isolated and purified from 9-day-old rats postnatal. Then serial ultra-thin slices were prepared and every single spermatogonium was observed and photographed by a Tecnai G2 12 TEM with magnification of 5800×, which were all inputted to a computer. After systemic numbering, we aligned the serial images by the naked eye and reconstructed the 3D image in Amira 3.1(TGS).Results A total of 7741 spermatogonia pictures in 120 serial sections were obtained. A spermatogonium which has 12 serial slices images was selected for 3D reconstruction. We reconstructed a part of cell stereostructure, furthermore internal and external structures of the model could be observed from any angles.Conclusion Within the help of Amira 3.1(TGS) and other softwares, 3D reconstructions of spermatogonia could be performed in PC-Windows platform of a common PC. Nevertheless, more convenient computer aided registration method should be raised, and the number of ultra-thin serial slices should be enriched for more abundant cell stereostructure information.PartⅣThree-dimensional reconstruction and virtual-reality technology facilitate living donor nephrectomyObjective To find a handy way to display the anatomy of the renal arterial and venous systems of potential donors on PC-Windows platform.Methods Dual-phase serial computed tomography (CT) images from ten living-related donors were loaded into TGS Amira 3.1 running on a PC with Windows XP. Image crop, segmentation and 3D reconstruction followed.Results We got the video clips as well as the 3D images of donor kidneys and blood vessels. According to the display which was confirmed by latter intraoperative findings, eight donors had single renal vessels, one had a left accessory renal artery and one had a right accessory renal artery.Conclusion This project offered a new approach to evaluate the renal vessel anatomy in living donor kidney transplantation, and it was favorable for accuracy and popularizing. Compared with expensive imaging workstations, it was much easier to be learned and manipulated by clinicians, and the clinicians can do their own interpretations according to the raw image data, it absolutely should be favorable for the accuracy of operation.