刺参体腔细胞的电镜制样技术研究

Study on the Electron Microscopyc Sample Preparation Technology of Apostichopus japonicus coelomocytes

设计2种不同的固定方法,比较2种固定方法在刺参体腔细胞的扫描电镜制样和透射电镜制样中固定效果的优劣。选择能使体腔细胞充分分散的固定方法用于扫描电镜制样;而将能使体腔细胞凝聚成团的固定方法用于透射电镜制样。获得了令人满意的体腔细胞扫描和透射电镜照片。以多聚赖氨酸法、明胶法和聚乙烯醇缩甲醛法收集、黏附固定后的体腔细胞,实验结果显示:3种方法均可起到黏附体腔细胞的作用,通过探讨玻片黏附体腔细胞的有效方法,建立适用于海产无脊椎动物体腔细胞、血细胞、生殖细胞和原生动物、细菌等样品的扫描电镜制样方法。

Two kinds of different fixation methods are designed, and their merits and demerits are compared in SEM and TEM sample preparations of Apostichopus japonicus coelomocytes. With these methods the coelomocytes, can be spread around in the SEM sample preparation or assembled in the TEM sample preparation. Satisfying SEM and TEM photographs are obtained using these methods. The fixed coelomocytes are collected with three methods; the poly-L-Lysine method, the glutin method, and the Formvar method. The experimental results show that all these methods are effective in making the coelomocytes to adhere to each other. By finding an effective method that causes the coelomocytes to adhere to the coverslips, a procedure is established for SEM sample preparation of marine invertobrate coelomocytes, haemocytes, reproductive cells, protozoa, etc.