仿刺参体腔细胞染色方法的研究

Staining methods of coelomocytes in sea cucumber Apostichopus japonicus

通过改变固定时间、孵育时间、染色时间及温度、分色方式等染色条件，研究不同染料、不同染色条件对仿刺参Apostichopus japonlcus体腔细胞的染色效果，并以观察体腔细胞内的颗粒、细胞核、细胞质和伪足的着色情况和细胞整体轮廓清晰程度为标准，筛选了适用于仿刺参体腔细胞的理想染色方法。结果表明：25℃室温下，采用滴片制片法，用湿盒孵育15～30min，用甲醇固定1～2min，用φ（Wright’s）：φ（Giemsa）=1：1的混合液染色1～2min，水洗，用中性树胶封片，在此条件下对仿刺参体腔细胞的染色效果最好。

The morphology and classification of coelomocytes are considered as the foundations for study on immune system of sea cucumber Apostichopus japonicus. In this study, the results of coelomocytes staining were compared using different dyes including Giemsa, Wright＇s, Wright＇s - Giemsa at different conditions （ fixed time, culture time, staining time and temperature, and decoloration method and time）. The perfect staining method of coelomocytes in the sea cucumber was obtained on the criteria of the staining of granules, nuclei, cytoplasm and pseudopodium and the definition of cell outline, which included cell drop technique, incubating in wet chamber for 15 - 30 min at 25 ℃, fixing in methanol for 1 -2 min, staining in φ（Wright＇s） ： φ（Giemsa） = 1：1 for 1 -2 min, washing in water and sealing up with colophony.