Name: simple homemade tools to handle fruit flies&#8212;drosophila melanogaster
Uploaded: 2019-07-24T13:30:00
Description: Watch this Scientific Journal Video about Simple Homemade Tools to Handle Fruit Fliesâ€”Drosophila melanogaster at JoVE.com

1. 准备工具和附件
<ol><li>提示/漏斗塞
	<ol>
<li>获得两个海绵插头(插头的直径必须略大于用于传输苍蝇的瓶的内径)。用加热的电焊铁在海绵塞的中心打一个洞。</li>
		<li>获得两个 1 mL 移液器尖端,用锋利的刀横向将一个切成两半,然后丢弃尖端。然后,从第二个移液器尖端上切下 1.5 厘米的尖端。将两个移液器尖端的残余与多用途粘合剂粘合在一起,使移液器尖端拉长(图1A<...

<ol>
	<li>Jennings, B. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drosophila+&#8211;+a+versatile+model+in+biology+&#38;+medicine.">Drosophila &#8211; a versatile model in biology &#38; medicine.</a> Materials Today. 14 (5), 190-195 (2011).</li><li>JoVE Science Education Database. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Biology I: yeast, Drosophila and C. elegans. An Introduction to Drosophila melanogaster. , (2018).</li><li>Ashburner, M., Roote, J., Sullivan, W., Ashburner, M., Hawley, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laboratory+Culture+of+Drosophila.">Laboratory Culture of Drosophila.</a> Drosophila Protocols. , (2000).</li><li>Greenspan, R. 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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Drosophila: A Guide to Species Identification and Use. , (2006).</li><li>Artiss, T., Hughes, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Taking+the+Headaches+Out+of+Anesthetizing+Drosophila:+A+Cheap+&#38;+Easy+Method+of+Constructing+Carbon+Dioxide+Staging.">Taking the Headaches Out of Anesthetizing Drosophila: A Cheap &#38; Easy Method of Constructing Carbon Dioxide Staging.</a> The American Biology Teacher. 69 (8), e77-e80 (2007).</li><li>Qu, W. -. H., Zhu, T. -. B., Yang, D. -. 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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。...

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。

<table>
	<tbody>
		<tr>
			<td>a pair of pliers</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>cordless drill driver</td>
			<td></td>
			<td></td>
			<td>max speed: 500 rpm</td>
		</tr>
		<tr>
			<td>electric soldering iron</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>file</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>funnel</td>
			<td></td>
			<td></td>
			<td>diameter of disk&#60;60mm</td>
		</tr>
		<tr>
			<td>ice box</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>insect pins</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>infrared thermometer</td>
			<td></td>
			<td></td>
			<td>HCIYET HT-830</td>
		</tr>
		<tr>
			<td>long cuff rubber gloves</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>mechanical pencils</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>medical gauze</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>microcentrifuge tube</td>
			<td></td>
			<td></td>
			<td>100 ul</td>
		</tr>
		<tr>
			<td>microwave oven</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Parafilm</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>peri dish</td>
			<td></td>
			<td></td>
			<td>internal diameter 60 mm</td>
		</tr>
		<tr>
			<td>pipette tips</td>
			<td></td>
			<td></td>
			<td>1 ml</td>
		</tr>
		<tr>
			<td>plastic film</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>plastic peri dish</td>
			<td></td>
			<td></td>
			<td>&#934;36 mm used to cover the empty vial</td>
		</tr>
		<tr>
			<td>point tweezers</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>protective work gloves</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>re-freezable hard icepacks</td>
			<td></td>
			<td></td>
			<td>26.5&#215;14.5&#215;2.5 cm or larger</td>
		</tr>
		<tr>
			<td>rubber air blower</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>snap cutter</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>soft drink bottle</td>
			<td></td>
			<td></td>
			<td>500 ml, internal diameter c.a. 65 mm</td>
		</tr>
		<tr>
			<td>sponge stopper</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>stainless steel sponges</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>tube brush</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>vial</td>
			<td></td>
			<td></td>
			<td>&#934;34 mm &#215; 90 mm</td>
		</tr>
	</tbody>
</table>

simple homemade tools to handle fruit flies&#8212;drosophila melanogaster

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。

T-和F-阻子被开发为一套简单的工具,可以调整和用于任何飞行转移活动。将苍蝇从旧文化转移到几种新鲜文化中,包括移除新鲜小瓶的插头,用F-塞器替换它们,然后轻敲旧小瓶中的苍蝇,快速取下其插头,并用T塞器替换。如果旧食物是紧凑的,那么重要的是翻转旧小瓶,并将T-止口的尖端插入F塞的开口,然后轻敲苍蝇进入新鲜的小瓶。然后,更换 T 和 F 塞器并重新插入小瓶。如果旧?...

Watch this Scientific Journal Video about Simple Homemade Tools to Handle Fruit Fliesâ€”Drosophila melanogaster at JoVE.com

Simple Homemade Tools to Handle Fruit Flies&#8212;Drosophila melanogaster

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。

简单的自制工具处理果蝇*果蝇黑色素

The fruit fly, Drosophila melanogaster, is widely used both in biological research and biology education. Handling adult flies is common but difficult in ...

转移和固定果蝇可能具有挑战性。使用这些技术和工具，您可以使常规飞行传输和固定更容易。制造这些工具所需的材料价格低廉，易于获得。咨询和教学或研究实验室可以让他们轻松。在开始此过程之前，使用电动搅铁在两个海绵塞的中心打一个洞，该孔大于用于传输苍蝇的小瓶的内部直径。接下来，用锋利的刀将一毫米小管尖横向切成两半，将第二个一毫米微管尖端的尖端切开1.5厘米。将两个移液器尖端的碎片与通用粘合剂粘合在一起。然后将拉长的微管尖端插入一个海绵塞中，并将漏斗插入另一个海绵塞中，然后用单独的 100 微升微离心管盖住移液器尖端。要准备微插针，请选择与符合人体工程学机械铅笔铅填充直径的昆虫针，并使用一对钳子切断针脚的独眼末端。然后将切口文件平放，然后用修改后的针脚替换铅笔中的引线。要准备一个果蝇蛋收集箱，使用捕捉切割机切割软塑料500毫升饮料瓶约2/3下瓶，并使用胶带固定卡纸条周围的60毫米直径的苹果汁板。要将成年苍蝇转移到小瓶之间，请仔细检查漏斗塞中的漏斗杆和尖端塞中的移液器尖端。轻点第一瓶中的苍蝇，用尖端塞子更换插头。用漏斗塞子插入第二个小瓶，并将第一个小瓶放在第二个小瓶上，以便将尖端塞的移液器尖端插入漏斗塞的漏斗开口。敲开第一个小瓶的边缘，让苍蝇从移液器尖端滑出，并通过漏斗的茎滑入第二个小瓶。如果第一瓶中的任何旧食物变得不那么紧凑，当小瓶倒置时掉落，将第二小瓶倒置在第一小瓶上，让苍蝇爬进第二小瓶。转移所需苍蝇数量后，将尖端塞从漏斗塞接器中分离，然后用原始插头更换尖端塞。然后用适当的插头更换第二瓶中的漏斗塞。对于成年果蝇通过冷却固定，放置一个可重新冷冻的冰袋，在零下80摄氏度下冷藏至少24小时，在室温下，并在碎冰中冷却空小瓶。20分钟后，稍微润湿了一块无菌医用纱布，比冰袋略小，有自来水。将纱布放在冰袋表面，使纱布紧粘附在冰袋表面。接下来，使用一块吸水纸擦拭冰冷空瓶中的水，然后将需要固定成根的成年苍蝇转移到小瓶中。当所有的苍蝇都转移后，盖住冰瓶，将小瓶敲碎冰两到三次，将所有苍蝇敲击到小瓶的底部。苍蝇会立即被固定。将小瓶放回冰上一分钟，然后将苍蝇倒在覆盖着冰袋的纱布上。然后用画笔来展开重叠的苍蝇。在立体显微镜下检查固定苍蝇，从解剖显微镜上取下舞台夹，用一块塑料薄膜覆盖舞台。将冰袋放在舞台上，并打开显微镜的冷光源。然后聚焦立体显微镜，移动冰袋，直到冰飞可以清楚地看到。观察苍蝇后，通过机翼将每只苍蝇转移到一个新的文化小瓶中。微波牺牲成年苍蝇计数，排序，或丢弃，转移感兴趣的成年苍蝇到空小瓶，并覆盖小瓶与培养皿。接下来将小瓶倒放在微波炉内，并在标准温度下加热小瓶80秒。死苍蝇会掉到培养皿里。戴上防护工作手套，将苍蝇倒入白色纸卡上，并在立体显微镜下转移卡，用先前准备的微切除针头进行检查和/或计数。苍蝇可以在考试结束时被丢弃。对于瓶形鸡蛋收集笼转移，插入改良的鸡蛋收集笼瓶到苹果汁板。使用石蜡薄膜条密封两个组件周围的接头。接下来用漏斗塞塞笼，将培养小瓶倒置到塞子上。然后将苍蝇加入笼子，通过漏斗塞子，然后用培养小瓶塞塞住笼子。要更换食物，请将苍蝇转移到空小瓶中，然后再用新盘子更换苹果汁盘。然后把苍蝇从小瓶传回笼子。在鸡蛋收集结束时，将苍蝇转移到空小瓶中，然后再将苍蝇转移到几个新的培养小瓶中。使用这种经过改良的冷麻醉方法，苍蝇在冰袋上保持不动，吸收冷凝水的医用纱布使苍蝇保持干燥，并可以轻松对冷冻苍蝇进行表型或处理。在25摄氏度的室温下，冰冷硬冰袋的表面温度急剧上升，从零下19摄氏度到零下2摄氏度，在20分钟内，迅速到达一个高原，对老苍蝇和新孵化的苍蝇都是安全的。与过度空化或深度冷冻相比，微波介电加热是一种更快、更方便杀死成年苍蝇的剂。与过度以太苍蝇类似，微波苍蝇从身体以直角伸展翅膀。一般来说，被微摇杀死的飞尸体的重量明显比被乙醚或冷冻杀死的飞尸体轻，但热量不会扭曲体型，而且尸体不会变得脆脆或笨拙。在将纱布压在冰袋上之前，请务必从湿润的纱布中挤出尽可能多的水，并确保纱布紧粘附在冰袋上。在将苍蝇转移到小瓶之前，将冷凝水从冰冷的空瓶上擦掉也很重要。

Research

JoVE Journal

Biology

Studying Aggression in Drosophila (fruit flies)

Studying Aggression in Drosophila fruit flies

学习在果蝇的侵略（果蝇）

Aggression is an innate behavior that evolved in the framework of defending or obtaining resources.  This complex social behavior is influenced by ...

科研

生物学

Using Micro-Electro-Mechanical Systems (MEMS) to Develop Diagnostic Tools

Using Micro-Electro-Mechanical Systems MEMS to Develop Diagnostic Tools

使用微机电系统（MEMS）开发诊断工具

Homemade Site Directed Mutagenesis of Whole Plasmids

自制的网站定向诱变整个质粒

Site directed mutagenesis of whole plasmids is a simple way to create slightly different variations of an original plasmid. With this method the cloned ...

Dissection of Oenocytes from Adult Drosophila melanogaster

Dissection of Oenocytes from Adult Drosophila melanogaster

从成人Oenocytes剖析果蝇</em

In Drosophila melanogaster, as in other insects, a waxy layer on the outer surface of the cuticle, composed primarily of hydrocarbon compounds, provides ...

Insulin Injection and Hemolymph Extraction to Measure Insulin Sensitivity in Adult Drosophila melanogaster

Insulin Injection and Hemolymph Extraction to Measure Insulin Sensitivity in Adult Drosophila melanogaster

注射胰岛素和血淋巴的提取来衡量成人胰岛素敏感性果蝇</em

Conserved nutrient sensing mechanisms exist between mammal and fruit fly where peptides resembling mammalian insulin and glucagon, respectively function ...

Isolation and Biophysical Study of Fruit Cuticles

果实角质层分离和生物物理研究

The cuticle, a hydrophobic protective layer on the aerial parts of terrestrial plants, functions as a versatile defensive barrier to various biotic and ...

Fruit Volatile Analysis Using an Electronic Nose

果实挥发分析，利用电子鼻

Numerous and diverse physiological changes occur during fruit ripening, including the development of a specific volatile blend that characterizes fruit ...

Measurement of Lifespan in Drosophila melanogaster

Measurement of Lifespan in Drosophila melanogaster

寿命测量<em&gt;果蝇</em

Aging is a phenomenon that results in steady physiological deterioration in nearly all organisms in which it has been examined, leading to reduced ...

High-throughput Functional Screening using a Homemade Dual-glow Luciferase Assay

使用自制双辉光萤光素酶检测的高通量筛选功能

We present a rapid and inexpensive high-throughput screening protocol to identify transcriptional regulators of alpha-synuclein, a gene associated with ...

Simple Method for Fluorescence DNA In Situ Hybridization to Squashed Chromosomes

Simple Method for Fluorescence DNA In Situ Hybridization to Squashed Chromosomes

对于荧光DNA的简易方法<em&gt;原位</em&gt;杂交来压扁染色体

DNA in situ hybridization (DNA ISH) is a commonly used method for mapping sequences to specific chromosome regions. This approach is particularly ...