简单的自制工具处理果蝇*果蝇黑色素
Summary
这里描述的是使用几个自制的工具转移,冷却,杀死成年果蝇,以及清洁玻璃养殖瓶和收集鸡蛋。这些工具易于制作,在处理果蝇方面相当有效。
Abstract
果蝇,果蝇黑色素,在生物学研究和生物学教育中被广泛应用。处理成年苍蝇是常见的,但在实践中很难,因为成年苍蝇飞行。这里演示的是如何使一些简单和具有成本效益的工具,以解决在处理果蝇的难题。泡沫塞中的孔被制成,移液器尖端或漏斗插入孔中。苍蝇然后只向一个方向移动到移液器尖端/漏斗组合,允许有效控制成人果蝇进入或流出小瓶。现有的协议已经修改,通过在碎冰中冷却并转移到寒冷、坚硬的冰袋表面来冷却苍蝇。冰袋上覆盖着一块医用纱布,在立体显微镜下检查时,使固定苍蝇远离冷凝水。苍蝇最终被安乐死以计数和分类,或被丢弃微摇。还开发了用于收集鸡蛋的瓶形笼子,以及用于清洁玻璃培养瓶的省力装置和附带的协议。
Introduction
果蝇,果蝇黑色素,是一种模型生物,广泛用于生物研究和生物学教育研究的广泛课题1,2。处理果蝇的基本问题是成年人从小瓶转移到小瓶和苍蝇的固定,使他们更容易处理,因为所有成年人(除了一些突变体3,4)可以飞行。
按照惯例,研究人员通过拿着两个小瓶口对口,轻击苍蝇或让苍蝇飞到另一个小瓶,然后分离和重新插入两个小瓶4,将苍蝇从一个小瓶转移到另一个小瓶。显然,这需要打开两个直径相同的小瓶,并且很难控制转移的苍蝇数量。同时,这需要快速的双手来完成工作,而逃避流浪苍蝇可能导致实验室或教室出现问题。在果蝇实验中,将特级处女苍蝇或雄蝇添加到已经准备好的十字架上是另一项例行任务。按照惯例,在添加额外的苍蝇之前,苍蝇必须固定在十字架小瓶中。
成人果蝇通常用醚、CO2或冷却5麻醉。与醚和CO2接触相比,冷却剂是固定成年果蝇的最经济有效的剂,对苍蝇和研究人员(尤其是年轻学生)的危害最小6,7。然而,在寒冷的表面或室内持续凝结的水会使苍蝇变湿。很难确定湿苍蝇的表型,在操作8、9中很容易损坏。这阻止了冷却方法得到更广泛的接受。
飞转工具和飞行冷却方法已经描述之前10。据报道,一种经过改良的冷却麻醉技术对果蝇实验是安全、可靠和可行的。本文还介绍了 1) 杀死成年人计数、分类或丢弃的方法,2) 清洁玻璃培养瓶的省力设备和规程,以及 3) 收集鸡蛋的简单笼子。此处描述的易于设计和经济高效的工具可用于解决飞行处理的难题,这些方法已经过测试,并且被证明为经验丰富的新手研究人员可靠、可靠且易于操作。
Protocol
Representative Results
Discussion
本文介绍了一些用于处理果蝇饲养和实验中涉及的基本活动的自制工具。这些工具很简单,但相当有效。实际上,任何实验室都可以轻松制作这些工具,而研究或教学实验室不需要找到现成的替代方案,而这种替代方案可能在当地不可用。
苍蝇转移是果蝇实验中最常见的做法和艰巨任务。不幸的是,到目前为止,还没有描述转移工具3,4,12,17在这里,T-和F-塞。…
Disclosures
The authors have nothing to disclose.
Acknowledgements
没有
Materials
| a pair of pliers | |||
| cordless drill driver | max speed: 500 rpm | ||
| electric soldering iron | |||
| file | |||
| funnel | diameter of disk<60mm | ||
| ice box | |||
| insect pins | |||
| infrared thermometer | HCIYET HT-830 | ||
| long cuff rubber gloves | |||
| mechanical pencils | |||
| medical gauze | |||
| microcentrifuge tube | 100 ul | ||
| microwave oven | |||
| Parafilm | |||
| peri dish | internal diameter 60 mm | ||
| pipette tips | 1 ml | ||
| plastic film | |||
| plastic peri dish | Φ36 mm used to cover the empty vial | ||
| point tweezers | |||
| protective work gloves | |||
| re-freezable hard icepacks | 26.5×14.5×2.5 cm or larger | ||
| rubber air blower | |||
| snap cutter | |||
| soft drink bottle | 500 ml, internal diameter c.a. 65 mm | ||
| sponge stopper | |||
| stainless steel sponges | |||
| tube brush | |||
| vial | Φ34 mm × 90 mm |
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