我们感谢美国深谷市先生、田硕先生和 t. Ishino 先生帮助进行了这些实验。这项研究得到了日本促进科学学会 (15K14755) 的科学研究资助。

1. 飞机厂的建造
<ol><li>
飞机磨机装置的施工
	<ol>
<li>从针 (金属部分:40 毫米长度和直径0.25 毫米) 切断塑料部分; 塑料部分: 长度为22毫米, 直径为2毫米) 带钳 (图 3)。</li>
		<li>用环氧树脂胶粘剂 (图 3) 在十字的形状上用未经处理的针固定这根针, 将其称为飞行磨臂和轴向针。 注: 对于轴向针, 未经处理的一侧应为底部。飞行磨房胳膊的被揭露的末端是为胶合甲虫 (图 1B和图 3)。</li>
	</ol>
</li>
	<li>
基地建设
	<ol>
<li>通过锤击钉子以防止轴向水平滑动 (图 4), 在薄薄的不锈钢金属板 (5 厘米 x 5 厘米) 表面上制作一个小酒窝。 注意: 金属板的实际尺寸不重要, 另一种材料是可能的, 但避免使用任何软材料;否则, 针会卡住, 防止磨机旋转。</li>
		<li>用胶带将木板上的金属板 (木制底座) 放置并固定。</li>
		<li>弯曲一个钢板, 使其双 L 形 (图 1C和图 2A)。 注: 用 L 型金属板固定墙上的家具是很方便的。另一种方便的观点是, 支持使用这种板块的是, 板块已经有很多孔。孔用于拧紧, 也可以修复一个捕捉按钮 (图 1A和图 4)。</li>
		<li>通过切割一次性塑料吸管的尖端 (高度 = 1 厘米, 外径 (外径) = 4 毫米, 内径 (id) = 2 毫米) 来制造圆柱形, 用于引导轴向针 (图 2A和图 4)。</li>
		<li>将双 L 形板和金属板上的气缸固定 (图 2A和图 4)。</li>
	</ol>
</li>
	<li>
传感设备的建设
	<ol>
<li>弯曲金属板使其 L 形, 使一个顶部板。 注: 使用 L 型金属板固定在墙上的家具是方便的 (图 5B-C)。如果是这样, 则可以跳过此步骤。</li>
		<li>在顶板上放置一个小的金属帽 (长度为5毫米, 直径为1毫米) (图 2D-E,图 4,图 5A)。 注意: 作为一个上限, 我们使用了一个捕捉按钮。它穿过 L 形板上的一个孔 (图 4)。</li>
		<li>修复 L 型板上的照片传感器 (图 4和图 5A)。拧紧 L 形板上传感器的电路基板以节省空间 (图 2D-E,图 4)。</li>
		<li>将红外线 led (150 兆瓦) 粘附在一个小磁铁上, 与 LED 的电路衬底 (图 1A和图 2A) 粘合。</li>
		<li>将 LED (150 兆瓦) 放在照片传感器下方的基板上 (图 1A和图 2A)。</li>
	</ol>
</li>
	<li>
施工支架
	<ol>
<li>弯曲金属板使其 L 形。 注: 使用 L 型金属板固定在墙上的家具是方便的 (图 5B-C)。如果是这样, 则可以跳过此步骤。</li>
		<li>用螺丝固定木板 (木墙) 上的盘子 (图 1C,图 4,图 5B)。木板的高度不重要, 这项研究是7厘米。</li>
	</ol>
</li>
	<li>
连接电缆
	<ol>
<li>通过普通电缆将照片传感器连接到 a/d 转换器的模拟输入通道。 注: 如果所有电缆都捆绑并固定在 L 形板上 (图 5B-D), 这是很有帮助的, 因为凌乱的工作区经常会防止整个实验中的细微操作。</li>
		<li>通过 USB 电缆将 a/d 转换器连接到个人计算机 (PC)。</li>
	</ol>
</li>
</ol>2. 实验程序
<ol><li>收集所有新出现的P. quercivorus成人从死的栎 crispula (Fagales: 壳斗科) 树在上午 (7-9 am) 的一天的实验将进行。 注意: 不要使用在前一天收集的甲虫。每天有超过100只甲虫出现, 新出现的甲虫每天都被检查。有关收集甲虫的详细方法, 请参阅参考16 。</li>
	<li>把甲虫放在冰上麻醉。避免把甲虫弄湿;否则, 将很难完成以下过程。在冰上执行所有后续的步骤。</li>
	<li>将一小部分的即时胶水 (湿乎乎胶) 的一部分, 在甲虫的 pronotum 与磨臂, 并保持磨臂与 pronotum 接触。 注: 如果单独使用胶水, 湿乎乎胶水会慢慢干燥。然而, 当两个组分混合 (材料表) 时, 这种胶水的作用很快。另一组分 (液体胶) 将在下一步使用。</li>
	<li>使用细针或棍子添加少量的胶水 (液体胶水) 的其他成分。确保机翼没有胶水 (图 1B)。液体胶用于促进湿乎乎胶的硬化。</li>
	<li>通过使用磁铁将 l 形板 (顶板) 放在另一 l 形板上, 将十字形针调整成机架 (图 6)。在调整针顶板高度时, 只需滑动顶部板即可。将轴向针的上尖端插入顶板上的 "捕捉" 按钮的孔 (图 5A), 然后将另一个笔尖放到基板上的导轨上 (图 6)。</li>
	<li>调整传感器下 IR 指示灯的位置。</li>
</ol>3. 获取和分析数据
<ol><li>将放大后的输出信号从照片传感器中记录下来, 并通过 a/d 转换器将其存储在 PC 中, 方法是使用具有1000磅/秒采样率的商用软件 (图 7A) (用于 a/d 转换器和软件、材料表)。</li>
	<li>启动软件 DAQFActoryExpress。</li>
	<li>在工作区窗口中, 单击日志记录图标上的十字 (+) 标记。</li>
	<li>右键单击日志记录集名称, 然后选择 "开始日志记录集"。 注意: 软件继续记录和保存数据。</li>
	<li>要停止录制, 请右键单击日志记录集名称, 然后选择 "结束日志记录集" 以保存. csv 文件。</li>
	<li>仅在记录的电压超过阈值 (0.5 V) 的情况下, 通过检测时间来提取在 IR LED 上方的飞行机臂的传递时间。 注意: 由于某些软件 (例如, Excel) 可以读取创建的. csv 文件, 所以根据研究的目的使用熟悉的软件。如有必要, 请下载通过 Github、https://github.com/HidetoshiIkeno/FlightMill 提供的定制程序。有关我们的程序的详细信息以及使用该程序的说明, 请参阅与主程序一起附带的自述文件。</li>
</ol>

<ol>
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作者没有什么可透露的。

我们开发了一个低成本, 易于建设, 紧凑的飞行磨机, 为小昆虫, 如P. quercivorus (4-5 毫米的身体长度和4-6 毫克的体重)。我们的飞行厂只包括普通的项目, 如针, 红外线 LED, 照片传感器, 瞬时胶水等, 不需要任何复杂的, 昂贵的, 或稀有的项目, 如计算机控制的电气设备。这样就可以方便快捷地收集必要的项目并降低实验成本。事实上, 每台机组的成本仅为1000日元 (约10美元、8欧元或7英镑) (?...

动物为了寻找食物和伴侣, 迁徙了很长的距离。迁徙的动物有时可能携带不良的同伴。雌豚草甲虫,鸭嘴兽 quercivorus (村山富市), 是一个已知的真菌病原体, Raffaelea quercivora Kubono 和新 Ito。这种病原体导致壳斗科树 (日本橡木枯萎) 的大量死亡和高水平的死亡率1。自1980年以来, 这种疾病一直在日本蔓延, 并已成为一个严重的问题2。
quercivorus是一种小昆虫 (体长4-5 毫米, 体重4-6 毫克), 每年的疾病扩展表明, 它们能够飞到几公里3,4。雄quercivorus定位一棵寄主树, 并释放一个聚合信息素, 吸引男性和女性5。因此, 宿主树受到 conspecifics 的大规模攻击, 最终死亡。在着陆后, 雄鸟在树内钻孔, 一个由信息素吸引的雌性进入隧道, 产卵。孵化的quercivours在隧道中生长, 直到成年。成人出现并分散, 以找到新的主机。因此, 疾病的扩展可能与这种甲虫的迁徙能力有关。然而, 甲虫能飞的程度仍然不明。此外, 雌性比雄性大6 (女性: 4.6 毫米, 雄性: 4.5 毫米) 和雄性甲虫搜索目标树, 进入树内的隧道, 然后吸引雌性。考虑到这些性别差异在身体大小和飞行的作用在他们的生活中, 性区别可能存在于飞行能力, 但能力的区别仍然不清楚。
一般来说, 由于迁徙区域的范围很广, 测量野外迁移能力, 特别是飞行能力, 是极其困难的。在60年7、8、9、10、11、12等拴系条件下的实验室测量了迁移能力, 例如一个飞行磨机系统;,13. 飞行磨机系统表明, 有些昆虫具有长距离飞行的能力。例如, 山松树甲虫的最长的飞行距离在一个飞行磨房是24公里14, 并且啮 planipennisi杨最大地飞行了7公里15。虽然飞行磨房是通常可利用的工具, 生物化验与活动物经常导致相当大个体区别。为了克服这一点, 许多测量, 重复多次, 需要得到可靠的估计平均扩散能力。因此, 应同时使用多个个人来快速收集足够数量的数据。然而, 同时实验需要一个更大的空间, 多个实验设置, 并比单一测量系统更昂贵。因此, 该机组必须低成本, 应易于建设与一般可用的项目, 并紧凑的规模。此外, 实验过程不应复杂或需要熟练的操作员。
在这项研究中, 我们组装了一个小型, 低成本的飞行厂 (图 1和图 2), 可以很容易地用于实验, 并测量飞行能力的豚草甲虫, P. quercivorus.

<table><tbody><tr><td>needle</td><td>Seirin</td><td>J type No. 5 x 40 mm</td><td></td></tr><tr><td>epoxy resin adhesive</td><td>Konishi</td><td>#16113</td><td></td></tr><tr><td>metal plate</td><td></td><td></td><td>from a home improvement store</td></tr><tr><td>disposable plastic pipette</td><td></td><td></td><td>from a home improvement store</td></tr><tr><td>snap button</td><td></td><td></td><td>from a craft store</td></tr><tr><td>IR sensor</td><td>Hamamatsu Photonics</td><td>S7136</td><td></td></tr><tr><td>IR LED</td><td>OptoSupply</td><td>OSIR5113A</td><td>150 mW</td></tr><tr><td>custom-made program</td><td></td><td></td><td>downloadable from Github.&lt;br/&gt; URL: https://github.com/HidetoshiIkeno/FlightMill</td></tr><tr><td>instant glue</td><td>Toagosei</td><td>31204</td><td></td></tr><tr><td>A/D converter</td><td>LabJack Co.</td><td>U3-HV</td><td></td></tr><tr><td>DAQ software</td><td>AzeoTech</td><td>DAQFactoryExpress</td><td>download from AzeoTech Web page.</td></tr></tbody></table>

measuring the flight ability of the ambrosia beetle, platypus quercivorus (murayama), using a low-cost, small, and easily constructed flight mill

豚草甲虫,鸭嘴兽 quercivorus (村山富市), 是真菌病原体的载体, 导致壳斗科树 (日本橡木枯萎) 的大量死亡。因此, 了解扩散能力可能有助于使诱捕/树移除的努力更有效地预防这种疾病。在这项研究中, 我们测量飞行速度和持续时间, 并估计飞行距离的甲虫使用新开发的飞行厂。该机组成本低, 体积小, 使用常用的物品建造。飞行磨机臂及其垂直轴组成一根细针。甲虫标本用速溶胶水粘在手臂的一个尖端上。另一个尖端是厚的由于被塑料覆盖, 从而有助于检测手臂的旋转。臂的革命是由安装在红外线 led 上的照片传感器检测到的, 当手臂通过 led 上方时, 输出电压的变化就表明了这一点。照片传感器连接到一台个人电脑, 输出电压数据以1赫的采样速率存储。通过使用这个飞行磨机进行实验, 我们发现quercivorus至少能飞27公里。由于我们的飞行厂包括便宜和小的普通项目, 许多飞行磨房可以同时被准备和使用在一个小实验室空间。这使实验者能够在短时间内获得足够数量的数据。

在这些实验中, 大约50% 的甲虫被应用到飞机上, 显示了一个或多个革命。当塑料部件通过传感器和 LED 之间的虚线时, 记录的电压从大约 0 v 变到约 6.5 v, 而传递的持续时间在10-20 毫秒以内, 这取决于飞行速度。因此, 像一场革命一样, 一个类似的峰值电压变化被观察到 (图 7B)。我们定义的飞行, 当飞行磨房胳膊旋转,即, 电压超出了门限 (0.5 V), 不管...

Watch this Scientific Journal Video about Measuring the Flight Ability of the Ambrosia Beetle, Platypus Quercivorus Murayama, Using a Low-Cost, Small, and Easily Constructed Flight Mill at JoVE.com

Measuring the Flight Ability of the Ambrosia Beetle, Platypus Quercivorus Murayama, Using a Low-Cost, Small, and Easily Constructed Flight Mill

利用低成本、小型、易于建造的Quercivorus测量豚草甲虫、鸭嘴兽的飞行能力 (村山富市)

我们开发了一个低成本和小型飞行厂, 建设与一般可用的项目, 并容易用于试验。利用这个仪器, 我们测量了一个豚草甲虫,鸭嘴兽 quercivorus的飞行能力。

The ambrosia beetle, Platypus quercivorus (Murayama), is the vector of a fungal pathogen that causes mass mortality of Fagaceae trees (Japanese oak wilt). ...

measuring-flight-ability-ambrosia-beetle-platypus-quercivorus

Universidad de Cartagena UDEC

Research

JoVE Journal

Environment

Measuring the Flight Ability of the Ambrosia Beetle, Platypus Quercivorus (Murayama), Using a Low-Cost, Small, and Easily Constructed Flight Mill

7.5K 次观看.  University of Hyogo. 我们开发了一个低成本和小型飞行厂, 建设与一般可用的项目, 并容易用于试验。利用这个仪器, 我们测量了一个豚草甲虫,鸭嘴兽 quercivorus的飞行能力。

视频: Measuring the Flight Ability of the Ambrosia Beetle, Platypus Quercivorus Murayama, Using a Low-Cost, Small, and Easily Constructed Flight Mill

Optimized Negative Staining: a High-throughput Protocol for Examining Small and Asymmetric Protein Structure by Electron Microscopy

Structural determination of proteins is rather challenging for proteins with molecular masses between 40 - 200 kDa. Considering that more than half of natural proteins have a molecular mass between 40 - 200 kDa1,2, a robust and high-throughput method with a nanometer resolution capability is needed. Negative staining (NS) electron microscopy (EM) is an easy, rapid, and qualitative approach which has frequently been used in research laboratories to examine protein structure and protein-protein interactions. Unfortunately, conventional NS protocols often generate structural artifacts on proteins, especially with lipoproteins that usually form presenting rouleaux artifacts. By using images of lipoproteins from cryo-electron microscopy (cryo-EM) as a standard, the key parameters in NS specimen preparation conditions were recently screened and reported as the optimized NS protocol (OpNS), a modified conventional NS protocol 3 . Artifacts like rouleaux can be greatly limited by OpNS, additionally providing high contrast along with reasonably high&#8208;resolution (near 1 nm) images of small and asymmetric proteins. These high-resolution and high contrast images are even favorable for an individual protein (a single object, no average) 3D reconstruction, such as a 160 kDa antibody, through the method of electron tomography4,5. Moreover, OpNS can be a high&#8208;throughput tool to examine hundreds of samples of small proteins. For example, the previously published mechanism of 53 kDa cholesteryl ester transfer protein (CETP) involved the screening and imaging of hundreds of samples 6. Considering cryo-EM rarely successfully images proteins less than 200 kDa has yet to publish any study involving screening over one hundred sample conditions, it is fair to call OpNS a high-throughput method for studying small proteins. Hopefully the OpNS protocol presented here can be a useful tool to push the boundaries of EM and accelerate EM studies into small protein structure, dynamics and mechanisms.

More than half of proteins are small proteins (molecular mass &#60;200 kDa) that are challenging for both electron microscope imaging and three-dimensional reconstructions. Optimized negative staining is a robust and high-throughput protocol to obtain high contrast and relatively high resolution (~1 nm) images of small asymmetric proteins or complexes under different physiological conditions.

优化负染：通过电子显微镜检查小和不对称蛋白结构的高通量协议

Structural determination of proteins is rather challenging for proteins with molecular masses between 40 - 200 kDa. Considering that more than half of ...

科研

环境科学

A New Application of the Electrical Penetration Graph (EPG) for Acquiring and Measuring Electrical Signals in Phloem Sieve Elements

Electrophysiological properties of cells are often studied in vitro, after dissociating them from their native environments. However, the study of electrical transmission between distant cells in an organism requires in vivo, artifact-free recordings of cells embedded within their native environment. The transmission of electrical signals from wounded to unwounded areas in a plant has since long piqued the interest of botanists. The phloem, the living part of the plant vasculature that is spread throughout the plant, has been postulated as a major tissue in electrical transmission in plants. The lack of suitable electrophysiological methods poses many challenges for the study of the electrical properties of the phloem cells in vivo. Here we present a novel approach for intracellular electrophysiology of sieve elements (SEs) that uses living aphids, or other phloem-feeding hemipteran insects, integrated in the electrical penetration graph (EPG) circuit. The versatility, robustness, and accuracy of this method made it possible to record and study in detail the wound-induced electrical signals in SEs of central veins of the model plant Arabidopsis thaliana1. Here we show that EPG-electrodes can be easily implemented for intracellular electrophysiological recordings of SEs in marginal veins, as well as to study the capacity of SEs to respond with electrical signals to several external stimuli. The EPG approach applied to intracellular electrophysiology of SEs can be implemented to a wide variety of plant species, in a large number of plant/insect combinations, and for many research aims.

A New Application of the Electrical Penetration Graph EPG for Acquiring and Measuring Electrical Signals in Phloem Sieve Elements

Electrical Penetration Graph (EPG) is a well-established technique for studying the feeding behavior of stylet-bearing insects. Here we show a new application of EPG as a non-invasive tool for the acquisition of intracellular electrophysiology recordings of sieve elements (SEs), the cells that form the phloem vasculature in plants.

在电气贯穿图（EPG）的韧皮部筛分子获取和测量电信号中的新应用

Electrophysiological properties of cells are often studied in vitro, after dissociating them from their native environments. However, the study of ...

The Benthic Exchange of O2, N2 and Dissolved Nutrients Using Small Core Incubations

The measurement of sediment-water exchange of gases and solutes in aquatic sediments provides data valuable for understanding the role of sediments in nutrient and gas cycles. After cores with intact sediment-water interfaces are collected, they are submerged in incubation tanks and kept under aerobic conditions at in situ temperatures. To initiate a time course of overlying water chemistry, cores are sealed without bubbles using a top cap with a suspended stirrer. Time courses of 4-7 sample points are used to determine the rate of sediment water exchange. Artificial illumination simulates day-time conditions for shallow photosynthetic sediments, and in conjunction with dark incubations can provide net exchanges on a daily basis. The net measurement of N2 is made possible by sampling a time course of dissolved gas concentrations, with high precision mass spectrometric analysis of N2:Ar ratios providing a means to measure N2 concentrations. We have successfully applied this approach to lakes, reservoirs, estuaries, wetlands and storm water ponds, and with care, this approach provides valuable information on biogeochemical balances in aquatic ecosystems.

The Benthic Exchange of O2, N2 and Dissolved Nutrients Using Small Core Incubations

Here, we present small core incubations for the measurement of sediment-water gas and solute exchange. These will provide reliable measurements of sediment-water exchange that assess the role of sediment in influencing biological and biogeochemical processes in aquatic ecosystems.

Ø的底栖交易所<sub&gt; 2</sub&gt;，N<sub&gt; 2</sub&gt;和营养盐使用小的核心孵化

The measurement of sediment-water exchange of gases and solutes in aquatic sediments provides data valuable for understanding the role of sediments in ...

Pretreatment of Lignocellulosic Biomass with Low-cost Ionic Liquids

A number of ionic liquids (ILs) with economically attractive production costs have recently received growing interest as media for the delignification of a variety of lignocellulosic feedstocks. Here we demonstrate the use of these low-cost protic ILs in the deconstruction of lignocellulosic biomass (Ionosolv pretreatment), yielding cellulose and a purified lignin. In the most generic process, the protic ionic liquid is synthesized by accurate combination of aqueous acid and amine base. The water content is adjusted subsequently. For the delignification, the biomass is placed into a vessel with IL solution at elevated temperatures to dissolve the lignin and hemicellulose, leaving a cellulose-rich pulp ready for saccharification (hydrolysis to fermentable sugars). The lignin is later precipitated from the IL by the addition of water and recovered as a solid. The removal of the added water regenerates the ionic liquid, which can be reused multiple times. This protocol is useful to investigate the significant potential of protic ILs for use in commercial biomass pretreatment/lignin fractionation for producing biofuels or renewable chemicals and materials.

The pretreatment of lignocellulosic biomass with protic low-cost ionic liquids is shown, resulting in a delignified cellulose-rich pulp and a purified lignin. The pulp gives rise to high glucose yields after enzymatic saccharification.

木质纤维素生物质预处理低成本离子液体

A number of ionic liquids (ILs) with economically attractive production costs have recently received growing interest as media for the delignification of ...

A Strain Gauge Monitor (SGM) for Continuous Valve Gape Measurements in Bivalve Molluscs in Response to Laboratory Induced Diel-cycling Hypoxia and pH

An inexpensive, laboratory-based, strain gauge valve gape monitor (SGM) was developed to monitor the valve gape behavior of bivalve molluscs in response to diel-cycling hypoxia. A Wheatstone bridge was connected to strain gauges that were attached to the shells of oysters (Crassostrea virginica). The recorded signals allowed for the opening and closing of the bivalves to be recorded continuously over two-day periods of experimentally-induced diel-cycling hypoxia and diel-cycling changes in pH. Here, we describe a protocol for developing an inexpensive strain gauge monitor and describe, in an example laboratory experiment, how we used it to measure the valve gape behavior of Eastern oysters (C. virginica), in response to diel-cycling hypoxia and cyclical changes in pH. Valve gape was measured on oysters subjected to cyclical severe hypoxic (0.6 mg/L) dissolved oxygen conditions with and without cyclical changes in pH, cyclical mild hypoxic (1.7 mg/L) conditions and normoxic (7.3 mg/L) conditions. We demonstrate that when oysters encounter repeated diel cycles, they rapidly close their shells in response to severe hypoxia and close with a time lag to mild hypoxia. When normoxia is restored, they rapidly open again. Oysters did not respond to cyclical pH conditions superimposed on diel cycling severe hypoxia. At reduced oxygen conditions, more than one third of the oysters closed simultaneously. We demonstrate that oysters respond to diel-cycling hypoxia, which must be considered when assessing the behavior of bivalves to dissolved oxygen. The valve SGM can be used to assess responses of bivalve molluscs to changes in dissolved oxygen or contaminants. Sealing techniques to better seal the valve gape strain gauges from sea water need further improvement to increase the longevity of the sensors.

A Strain Gauge Monitor SGM for Continuous Valve Gape Measurements in Bivalve Molluscs in Response to Laboratory Induced Diel-cycling Hypoxia and pH

Understanding the behavioral responses of bivalve suspension-feeders to environmental variables, such as dissolved oxygen, can explain some ecosystem processes. We have developed an inexpensive, laboratory-based, strain gauge monitor (SGM) to measure valve gape responses of oysters, Crassostrea virginica, to diel-cycling hypoxia and cyclical pH.

基于实验室诱导的昼夜循环缺氧和 pH 值的双壳软体动物连续气门目瞪口呆测量用应变计监测仪 (通用)

An inexpensive, laboratory-based, strain gauge valve gape monitor (SGM) was developed to monitor the valve gape behavior of bivalve molluscs in response ...

Using Flight Mills to Measure Flight Propensity and Performance of Western Corn Rootworm, Diabrotica virgifera virgifera (LeConte)

The western corn rootworm, Diabrotica virgifera virgifera (LeConte) (Coleoptera: Chrysomelidae), is an economically important pest of corn in the northern United States. Some populations have developed resistance to management strategies including transgenic corn that produces insecticidal toxins derived from the bacterium Bacillus thuringiensis (Bt). Knowledge of western corn rootworm dispersal is of critical importance for models of resistance evolution, spread, and mitigation. Flight behavior of an insect, especially over a long distance, is inherently difficult to observe and characterize. Flight mills provide a means to directly test developmental and physiological impacts and consequences of flight in the laboratory that cannot be obtained in field studies. In this study, flight mills were used to measure the timing of flight activity, total number of flights, and the distance, duration, and speed of flights taken by female rootworms during a 22-h test period. Sixteen flight mills were housed in an environmental chamber with programmable lighting, temperature, and humidity control. The flight mill described is of a typical design, where a flight arm is free to rotate about a central pivot. Rotation is caused by flight of an insect tethered to one end of the flight arm, and each rotation is recorded by a sensor with a time-stamp. Raw data are compiled by software, which are subsequently processed to provide summary statistics for flight parameters of interest. The most difficult task for any flight mill study is attachment of the tether to the insect with an adhesive, and the method used must be tailored to each species. The attachment must be strong enough to hold the insect in a rigid orientation and to prevent detachment during movement, while not interfering with natural wing motion during flight. The attachment process requires dexterity, finesse, and speed, making video footage of the process for rootworms of value.

Using Flight Mills to Measure Flight Propensity and Performance of Western Corn Rootworm, Diabrotica virgifera virgifera LeConte

Flight mills are important tools for comparing how age, sex, mating status, temperature, or various other factors may influence an insect&#8217;s flight behavior. Here we describe protocols to tether and measure the flight propensity and performance of western corn rootworm under different treatments.

使用飞行磨坊测量西部玉米根虫的飞行倾向和性能，二溴二恶十二药病毒病毒（LeConte）

The western corn rootworm, Diabrotica virgifera virgifera (LeConte) (Coleoptera: Chrysomelidae), is an economically important pest of corn in the northern ...

行为学

A Low-cost Method for Analyzing Seizure-like Activity and Movement in Drosophila

Video tracking systems have been used widely to analyze Drosophila melanogaster movement and detect various abnormalities in locomotive behavior. While these systems can provide a wealth of behavioral information, the cost and complexity of these systems can be prohibitive for many labs. We have developed a low-cost assay for measuring locomotive behavior and seizure movement in D. melanogaster. The system uses a web-cam to capture images that can be processed using a combination of inexpensive and free software to track the distance moved, the average velocity of movement and the duration of movement during a specified time-span. To demonstrate the utility of this system, we examined a group of D. melanogaster mutants, the Bang-sensitive (BS) paralytics, which are 3-10 times more susceptible to seizure-like activity (SLA) than wild type flies. Using this novel system, we were able to detect that the BS mutant bang senseless (bss) exhibits lower levels of exploratory locomotion in a novel environment than wild type flies. In addition, the system was used to identify that the drug metformin, which is commonly used to treat type II diabetes, reduces the intensity of SLA in the BS mutants.

A Low-cost Method for Analyzing Seizure-like Activity and Movement in Drosophila

Using a webcam and a combination of free and inexpensive software programs, locomotive patterns in Drosophila melanogaster can be analyzed to detect differences in the speed, distance, and time of various motor activities.

在分析癫痫样活动和运动的一种低成本方法<em&gt;果蝇</em

Video tracking systems have been used widely to analyze Drosophila melanogaster movement and detect various abnormalities in locomotive behavior. While ...

神经科学

Building an Enhanced Flight Mill for the Study of Tethered Insect Flight

Makerspaces have a high potential of enabling researchers to develop new techniques and to work with novel species in ecological research. This protocol demonstrates how to take advantage of the technology found in makerspaces in order to build a more versatile flight mill for a relatively low cost. Given that this study extracted its&#160;prototype from flight mills built in the last decade, this protocol focuses more on outlining divergences made from the simple, modern flight mill. Previous studies have already shown how advantageous flight mills are to measuring flight parameters such as speed, distance, or periodicity. Such mills have allowed researchers to associate these parameters with morphological, physiological, or genetic factors. In addition to these advantages, this study discusses the benefits of using the technology in makerspaces, like 3D printers and laser cutters, in order to build a more flexible, sturdy, and collapsible flight mill design. Most notably, the 3D printed components of this design allow the user to test insects of various sizes by making the heights of the mill arm and infrared (IR) sensors adjustable. The 3D prints also enable the user to easily disassemble the machine for quick storage or transportation to the field. Moreover, this study makes greater use of magnets and magnetic paint to tether insects with minimal stress. Lastly, this protocol details a versatile analysis of flight data through computer scripts that efficiently separate and analyze differentiable flight trials within a single recording. Although more labor-intensive, applying the tools available in makerspaces and on online 3D modeling programs facilitates multidisciplinary and process-orientated practices and helps researchers avoid costly, premade products with narrowly adjustable dimensions. By taking advantage of the flexibility and reproducibility of technology in makerspaces, this protocol promotes creative flight mill design and inspires open science.

This protocol uses three-dimensional (3D) printers and laser cutters found in makerspaces in order to create a more flexible flight mill design. By using this technology, researchers can reduce costs, enhance design flexibility, and generate reproducible work when constructing their flight mills for tethered insect flight studies.

为系留昆虫飞行研究建造增强型飞行磨机

Makerspaces have a high potential of enabling researchers to develop new techniques and to work with novel species in ecological research. This protocol ...

工程学

A Simple Flight Mill for the Study of Tethered Flight in Insects

Flight in insects can be long-range migratory flights, intermediate-range dispersal flights, or short-range host-seeking flights. Previous studies have shown that flight mills are valuable tools for the experimental study of insect flight behavior, allowing researchers to examine how factors such as age, host plants, or population source can influence an insects' propensity to disperse. Flight mills allow researchers to measure components of flight such as speed and distance flown. Lack of detailed information about how to build such a device can make their construction appear to be prohibitively complex. We present a simple and relatively inexpensive flight mill for the study of tethered flight in insects. Experimental insects can be tethered with non-toxic adhesives and revolve around an axis by means of a very low friction magnetic bearing. The mill is designed for the study of flight in controlled conditions as it can be used inside an incubator or environmental chamber. The strongest points are the very simple electronic circuitry, the design that allows sixteen insects to fly simultaneously allowing the collection and analysis of a large number of samples in a short time and the potential to use the device in a very limited workspace. This design is extremely flexible, and we have adjusted the mill to accommodate different species of insects of various sizes.

Flight in insects is influenced by a number of factors and the propensity to disperse is an important variable in understanding insect ecology and biological control strategies. We describe the construction and use of a simple, relatively inexpensive, and flexible flight mill for measuring parameters of tethered flight in insects.

一个简单的飞行磨的系留飞行的昆虫研究

Flight in insects can be long-range migratory flights, intermediate-range dispersal flights, or short-range host-seeking flights. Previous studies have ...

An Improved Method for Accurate and Rapid Measurement of Flight Performance in Drosophila

Drosophila has proven to be a useful model system for analysis of behavior, including flight. The initial flight tester involved dropping flies into an oil-coated graduated cylinder; landing height provided a measure of flight performance by assessing how far flies will fall before producing enough thrust to make contact with the wall of the cylinder. Here we describe an updated version of the flight tester with four major improvements. First, we added a &#34;drop tube&#34; to ensure that all flies enter the flight cylinder at a similar velocity between trials, eliminating variability between users. Second, we replaced the oil coating with removable plastic sheets coated in Tangle-Trap, an adhesive designed to capture live insects. Third, we use a longer cylinder to enable more accurate discrimination of flight ability. Fourth we use a digital camera and imaging software to automate the scoring of flight performance. These improvements allow for the rapid, quantitative assessment of flight behavior, useful for large datasets and large-scale genetic screens.

An Improved Method for Accurate and Rapid Measurement of Flight Performance in Drosophila

Here we describe a method for rapid and accurate measurement of flight performance in Drosophila, enabling high-throughput screening.

在对飞行性能的准确，快速测定方法的改进<em&gt;果蝇</em

Drosophila has proven to be a useful model system for analysis of behavior, including flight. The initial flight tester involved dropping flies into an ...

利用低成本、小型、易于建造的Quercivorus测量豚草甲虫、鸭嘴兽的飞行能力 (村山富市)

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