There are five developmental stages in the spermatogenesis , namely are primary spermatogonium , secondary spermatogonium , primary spermatocytes , secondaty spermatocytes and spermatids . the spermatids develop into sperm via the process of spermiogenesis . there exist cell junctions between two spermatids 电子显微镜观察表明：大眼鳜的精子发生和形成过程，经历了初级精原细胞，次级精原细胞，初级精母细胞，次级精母细胞和精子细胞阶段。
Except the incomplete maturation of spermatid nuclear and oocyte activation , idendification of a live spermatid is a pivotal procedure . it is difficult to distinguish round spermatids from other round cells such as spermatocytes , monocytes , polymorphonuclear leukocytes and so on 除精子细胞的核蛋白不完全成熟及卵子激活不足等因素外，如何正确选择存活精子细胞是个难题，如圆形精子细胞与精母细胞、单核细胞和多形核白细胞等其他圆形细胞的区分就比较困难。
In the first trial , combination of enzymatic digestion was used to prepare suspensions of spermatogenic cells from adult mouse testis , and then a modified discontinuous percoll gradient centrifugation method ( 15 % , 22 % , 30 % , 40 % , 50 % , 60 % ) was introduced to isolate spermatids from the cellular suspensions . the content of spermatids in each isolated fraction by percoll method was determined by morphology ( wright - giemsa stain ) and flow cytometry analysis , and the viability of spermatogenic cells was assessed by using eosin y exclusion test 在第一部分试验中，首先利用连续3次组合酶消化成年小鼠睾丸制备睾丸细胞悬液，然后经6层非连续percoll梯度离心法( 15 、 22 、 30 、 40 、 50和60 )分离，通过形态学和流式细胞术鉴定南京医科大学硕士学位论文各个percoll组分中精子细胞的含量，并以伊红y排斥试验测定细胞的存活率。
The first reports on successful clinical use of round and elongated spermatids for assisted reproduction in 1995 suggest the benefits that the technique of spermatid conception can bring to some patients suffering from otherwise untreatable types of male factor infertility . men with non - obstructive azoospermia ( noa ) can now be treated by using intra - oocyte round spermatid injection ( rosi ) or elongated spermatid injection ( elsi ) . spermatids can be retrieved from semen or from testis biopsy specimens 目前，完全不能产生精子的非梗阻性无精子症( non - obstmctiveazoospermia , noa )患者可采用卵胞质内圆形精子细胞注射( roundspermatidinjection , rosi )或长形精子细胞注射( elongatedspermatidinection , elsi )治疗，精子细胞可从精液或从睾丸活检组织中获取。
Nucleoli do not emerge in this process . the number of the organelles increase until secondary spermatocyte stage . mitochondria accumulate together , merging together with lysosomes and golgi bodies at the early spermatid stage , and finally the lamellar structure is formed , which forms the acrosome at last 在精子发生过程中，线粒体、内质网和核糖体逐渐增多，其中线粒体数目在次级精母细胞阶段达到顶峰，并形成线粒体区，精细胞早期核内出现膜性泡结构，同时次级溶酶体与高尔基体大量存在，这些细胞器共同形成片层复合体，并参与顶体的形成。
In the second trial , this modified discontinuous percoll gradient centrifugation method was introduced to isolate spermatids from the semen of fifteen male infertile patients . then the effect was identified by wright - giemsa stain , flow cytometry analysis , immunocytochemistry and fluorescence in situ hybridization ( fish ) . similary , the 22 % percoll fraction contained mostly haploid cells [ ( 91 . 85 ? 5 . 18 ) % ] ( p < 0 . 005 ) and the mean density in this fraction was ( 1 . 010 ? 0 . 786 ) x 105 / ml C法，对15例各种类型不育患者的精液细胞进行分离，并利用瑞姬染色法、流式细胞术、免疫细胞化学和荧光原位杂交oisffi等方法，从细胞形态特征、 dna倍体、细胞表面标i己与分化抗原，以及原位杂交信号的数目和位置结合细胞核特有的形态等方面加以鉴定。
The cellular localization of hse was done by using in situ hybridization , the results showed that primary spermtatocytes and spermatids in seminiferous tubular have positive signals . the subcellular localizations of hsei and hseii were identified by gfp fusion protein . hsei - gfp fusion protein was evenly distributing in cytoplasm , while hseii - gfp was not evenly distributing in cytoplasm and there were many bright spots in cytosol 用gfp高合蛋白的技术确定hsei和hsell在细胞中勺定位，结果显示hsei gfp均匀地分布于细胞浆中，而hsell0fp则分布不均匀，在胞浆有聚集成许多荧光信号很强的点。
Although with these methods , spermatids can be isolated manually under the microscope for immediate use , it is not possible to obtain large populations of purified spermatids . but repeated biopsies for retrieving spermatids can further compromise the already deficient seminiferous tubules of patients 然后在显微镜下从中手工选择精子细胞供立即穿刺使用，但不可能获得大量纯化的精子细胞，而反复活检获取精子细胞将进一步损伤患者已有缺陷的睾丸精曲小管( seminiferoustubules ) 。
Nucleoli exist from spermatocyte stage to early spermatid stage . in this process , the numbers of ribosomes , mitochondrions and endoplasmic reticulums increase until secondary spermatocyte stage , golgi apparatus emerges at the secondary spermatocyte stage , and then , these organelles change into lamellar complex which finally forms the acrosome 核糖体，线粒体和内质网等细胞器数量逐渐增多，到次级精母细胞阶段达到最大，在形成精子的过程中，上述细胞器与高尔基体及膜性泡共同分化或参与形成片层小体，并参与顶体的形成。
The number of mitochondrion is more less than the endoplasmic reticulum , and the smooth endoplasmic reticulum is the main kind of the endoplasmic reticulum ; golgi bodies and lysosomes emerge in the secondary spermatocyte stage . finally , these organelles change into pre - acrosome vesicles which become acrosome at last . sinopotamon chekiangense during the spermatogenensis , chronmatins condense at different level until middle spermatid stage 在整个发生过程中细胞器数量较少，内质网数目在各细胞器中所占比例最大，以滑面内质网为主，线粒体在初级精母细胞中最多，自次级精母细胞开始逐渐减少，高尔基体和溶酶体自次级精母细胞始出现，在发育过程中上述细胞器不断分化，在精细胞阶段形成前顶体腔，最后形成圆球形顶体。
