Encyclopedia of Experiments: Biology
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- 要固定苍蝇的石蜡嵌入,小心地推动麻醉的德罗索菲拉通过苍蝇领的缝隙,使他们的头均匀地放置在设备的顶部,同时仍然连接到身体的其余部分。对于组织固定,将衣领放在卡诺伊的溶液中。卡诺伊的是一种由乙醇、氯仿和醋酸组成的变性固定剂。
为了准备石蜡渗透的样品,通过分级乙醇浓度转移衣领,使组织脱水。 然后,将衣领转移到有机溶剂甲基苯甲酸酯中。接下来,将其放置在甲基苯甲酸酯和融化的石蜡溶液中加热,以确保石蜡保持熔化。 继续使用预热石蜡单独进行一系列精心设计的洗涤步骤,以去除甲基苯甲酸酯。
接下来,迅速将衣领放在模具中,倒入融化的石蜡以完全覆盖它。 石蜡硬化后,从模具中取出方块,使用剃须刀刀片修剪多余的石蜡。 将领子脱落,将一排嵌入式头留在石蜡块中。在示例协议中,我们将看到用于分析神经退行性表型的程序。
-首先, 首先,使用钳子将受试者从颈部飞入衣领。将所有头部对齐相同的方向,并确保头部或眼睛不会受到损伤。包括无眼正弦奥库利斯苍蝇在随机已知的位置,以便苍蝇的顺序可以很容易地识别在部分。 此外,如果感兴趣的苍蝇有光或白眼睛,线状红眼苍蝇每隔一段时间沿着条,以确保有足够的色素存在污渍幻灯片。
如果使用多个领子,则将苍蝇的顺序与领号一起记录。 完成衣领后,将其放入卡诺伊的溶液中3.5至 4 小时。 执行一系列乙醇、甲基苯甲酸酯和石蜡洗涤,为苍蝇组织嵌入做好准备。接下来,将衣领放入橡胶冰块托盘中,每节一个。
轻轻地将熔融的石蜡倒在衣领上,避免气泡,并允许它在一夜之间变硬。从托盘中取出包含衣领的石蜡块。使用剃须刀刀片将石蜡块从衣领上分离,轻轻折断衣领。 尸体将留在衣领中,而头部将留在石蜡块中。
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