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- 首先,将一块带狭缝的湿海绵放在培养皿上。接下来,用三卡因溶液麻醉成年鱼,并小心地将其转移到腹侧朝上进入狭缝中。将培养皿置于立体显微镜下,观察皮下跳动心脏的运动。
将 mincroinjection 针头与位于鳃盖和胸带之间的跳动心脏上方的体轴成 45 度角。轻轻地将针头穿过皮肤进入心包,心包是一层围绕心脏的薄膜。将注射混合物输送到心包中。
该镜头包含示踪染料,例如酚红,用于跟踪溶液的运动。从胸部抽出针头。立即将鱼转移到装有系统水的回收罐中,并监测它是否从麻醉中完全恢复。在下面的方案中,我们将进行胸腔内注射以输送测试化合物并检查其对心脏再生的影响。
- 用网捕捉一条成年鱼并将其转移到麻醉溶液中。一到两分钟后,当鱼停止游泳并且鳃盖的运动减少时,用塑料勺触摸鱼,以确保它不会对任何接触做出反应。接下来,快速小心地将鱼转移到湿海绵的凹槽中,腹侧朝上。鱼的头部应远离作员的惯用手。
在立体显微镜下,仔细观察鱼皮下跳动的心脏的运动。目测确定跳动心脏上方和腹侧软骨板定义的三角形中间的注射点。以相对于本体轴线 30 到 45 度的角度插入毛细管的尖端。用微毛细管的尖端轻轻穿透皮肤进入心包。
针头进入心包后,踩下显微注射器装置的踏板以完成注射。注射后,轻轻地从胸部抽出毛细血管,并立即将鱼转移到装有系统水的水箱中进行回收。监测鱼直到从麻醉中完全恢复。为了分析注射的效果,在所需的时间点收集心脏并准备进一步分析。
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