Name: the ingestion of fluorescent, magnetic nanoparticles for determining fluid-uptake abilities in insects
Uploaded: 2017-12-20T15:00:00
Description: Watch this Scientific Journal Video about The Ingestion of Fluorescent, Magnetic Nanoparticles for Determining Fluid-uptake Abilities in Insects at JoVE.com

この作品は国立科学財団 (NSF) によってサポートされていた許可なし。IOS 1354956。博士アンドリュー d. ウォーレン (鱗翅目、生物多様性、フロリダ州フロリダ大学自然史博物館・ マクガイア センター) 蝶の画像を使用する許可を感謝します。

1. 昆虫種、ソリューションおよび供給ステーションのセットアップの準備
注: 流体吸収能力、口器形態22,23に関する以前の研究の使用のため、キャベツ蝶 (モンシロチョウL.、シロチョウ科) は代表チョウ目の種として選択されます。家 (イエバエL., イエバエ科) や青いボトル蠅 (オオクロバエ vomitoria L., クロバ...

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昆虫の口器機能は形態学からみた歴史的に (e.g、鱗翅目のテング機能関連飲むわら25,26);。ただし、実験的証拠が組み込まれた最近の研究は、昆虫の口器と構造機能関係2,12,13の複雑さの私達の理解のパラダイム シフトで起因しました。,22,...

多くの昆虫のグループ (proboscises) の口器がある適応流体を供給するため、花の蜜、果実の腐敗など sap フロー (例:双翅目1、鱗翅目2膜翅目3)、木部 (半翅目4)、涙 (鱗翅目5) と血 (目, シラミ目6ノミ7、双翅目7、8半翅目、鱗翅目9)。流体を餌に昆虫の能力は生態系の健全性 (例えば受粉10)、病気伝達4,11生物2,12、および研究に関連します。収斂進化13。多彩な食料源、にもかかわらずいくつかの流体餌昆虫の中でテーマとマイクロやナノ液滴、液膜または多孔性の表面に限定されるが、液体の少量を取得する機能です。
流体食性昆虫 (すべての動物種の14,の15の 20% 以上) と様々 な食料源を餌に能力の膨大な多様性を与えられた彼らの餌を理解動作流体吸収機構の解明、多くの分野で重要であります。昆虫の口器の機能、たとえば、バイオミメティック技術、例えば少量の流体が用いられるものと同様の方法を使用しての取得などのタスクを実行できるマイクロ流体デバイスの開発の役割を果たしています。昆虫16。流体吸収メカニズムの研究における根本的な問題は、流体、昆虫のフィード方法のみを決定するが、メカニズムをサポートして実験的証拠を取得します。のみ動作を使用して (例えば、テング12,17プロービング) 流体の正常な吸収を確認しませんも、それがルートを決定するための手段を提供するため、供給のための指標が十分なこと流体の旅行昆虫を通過するとき。また、少量より流体の実験を実行する流体が制限リソース2,12自然の供給シナリオを表します。
位相コントラスト イメージング多孔性の表面12から液体の少量で蝶がどのようにフィードを評価するオオカバマダラ (オオカバマダラl.) を併用した x 線します。モナルカ蝶フード運河に小さな毛穴に閉じ込められた流体をもたらすテングに沿って毛細管を介して表皮突起 (背 legulae) の間にスペースを使用します。着信の流体は、成長し高原不安定12,18, 頭に吸引ポンプの作用で、蝶の腸を運ばれる液体の橋に崩壊食品管壁のフィルムを形成します。X 線位相コントラスト イメージング昆虫12,19,20,21内の流体の流れを可視化するための最適なツールですが、テクニックはすぐに利用できるより便利メソッドは、昆虫の能力吸収液の迅速な評価のために必要ですし、摂取します。
D. plexippusの供給メカニズム適用他の鱗翅目に、ハエ (双翅目)、(両方のグループが多孔性の表面の液体のフィード) かどうかを決定する Lehnertら13には、ここで詳細に報告されている多孔性の表面の液体の少量を餌に昆虫の能力を評価するための手法が適用されます。ここで説明したプロトコルは接液部を使用して研究と多孔質の表面、プール供給メカニズムのアドレスなど、他の研究方法論を変更できます。また、アプリケーションは、マイクロ流体システムと材料技術など、あらゆる分野に拡張します。

<table>
	<tbody>
		<tr>
			<td>20% sucrose solution</td>
			<td>Domino Sugar</td>
			<td></td>
			<td>Sugar needed to produce the sucrose solution with dH2O</td>
		</tr>
		<tr>
			<td>Phosphate Buffered Saline (PBS)</td>
			<td>Sigma-Aldrich</td>
			<td>P5493</td>
			<td>10X concentration diluted to 1X in dH2O for insect dissections</td>
		</tr>
		<tr>
			<td>Single depression concave slide</td>
			<td>AmScope</td>
			<td>BS-C6</td>
			<td>Slide is necessary for feeding stage setup</td>
		</tr>
		<tr>
			<td>Filter paper</td>
			<td>EMD Millipore</td>
			<td>NY6004700 (60 &#181;m)</td>
			<td>Nylon net filters and isopore filters needed to produce a porous surface for insect feeding</td>
		</tr>
		<tr>
			<td>Filter paper</td>
			<td>EMD Millipore</td>
			<td>NY4104700 (41 &#181;m)</td>
			<td>Nylon net filters and isopore filters needed to produce a porous surface for insect feeding</td>
		</tr>
		<tr>
			<td>Filter paper</td>
			<td>EMD Millipore</td>
			<td>NY3004700 (30 &#181;m)</td>
			<td>Nylon net filters and isopore filters needed to produce a porous surface for insect feeding</td>
		</tr>
		<tr>
			<td>Filter paper</td>
			<td>EMD Millipore</td>
			<td>NY2004700 (20 &#181;m)</td>
			<td>Nylon net filters and isopore filters needed to produce a porous surface for insect feeding</td>
		</tr>
		<tr>
			<td>Filter paper</td>
			<td>EMD Millipore</td>
			<td>NY1104700 (11 &#181;m)</td>
			<td>Nylon net filters and isopore filters needed to produce a porous surface for insect feeding</td>
		</tr>
		<tr>
			<td>Filter paper</td>
			<td>EMD Millipore</td>
			<td>TCTP04700 (10 &#181;m)</td>
			<td>Nylon net filters and isopore filters needed to produce a porous surface for insect feeding</td>
		</tr>
		<tr>
			<td>Filter paper</td>
			<td>EMD Millipore</td>
			<td>TETP04700 (8 &#181;m)</td>
			<td>Nylon net filters and isopore filters needed to produce a porous surface for insect feeding</td>
		</tr>
		<tr>
			<td>Filter paper</td>
			<td>EMD Millipore</td>
			<td>TMTP04700 (5 &#181;m)</td>
			<td>Nylon net filters and isopore filters needed to produce a porous surface for insect feeding</td>
		</tr>
		<tr>
			<td>Filter paper</td>
			<td>EMD Millipore</td>
			<td>RTTP04700 (1 &#181;m)</td>
			<td>Nylon net filters and isopore filters needed to produce a porous surface for insect feeding</td>
		</tr>
		<tr>
			<td>Iris microdissecting scissors</td>
			<td>Carolina Biological Supply Company</td>
			<td>623555</td>
			<td>Scissors used for dissections</td>
		</tr>
		<tr>
			<td>Insect pins (#1)</td>
			<td>Bioquip Products</td>
			<td>1208B1</td>
			<td>Pins used during dissections and feeding trials</td>
		</tr>
		<tr>
			<td>Extra-fine point dissecting forceps</td>
			<td>Carolina Biological Supply Company</td>
			<td>624684</td>
			<td>Dissecting equipment</td>
		</tr>
		<tr>
			<td>Leica M205 C Stereoscope</td>
			<td>Leica Microsystems</td>
			<td>M205 C</td>
			<td>Stereoscope used for dissections</td>
		</tr>
		<tr>
			<td>Inverted confocal microscope</td>
			<td>Olympus</td>
			<td>IX81</td>
			<td>Fluorescent microscope used to detect magnetic nanoparticles</td>
		</tr>
		<tr>
			<td>Fisherbrand PTFE Disposable Stir Bar</td>
			<td>Fisherscientific</td>
			<td>S68067</td>
			<td>Magnet used to detect nanoparticles</td>
		</tr>
		<tr>
			<td>Kimtech Science Kimwipes</td>
			<td>Kimberly-Clark Professional</td>
			<td>34155</td>
			<td>Tissues used to secure insects during feeding trials</td>
		</tr>
		<tr>
			<td>House fly (Musca domestica) pupae</td>
			<td>Mantisplace.com</td>
			<td></td>
			<td>insects for experiments</td>
		</tr>
		<tr>
			<td>Blue bottle fly (Calliphora vomitoria) pupae</td>
			<td>Mantisplace.com</td>
			<td></td>
			<td>insects for experiments</td>
		</tr>
		<tr>
			<td>Cabbage butterfly (Pieris rapae) larvae</td>
			<td>Carolina Biological Supply Company</td>
			<td>144102</td>
			<td>insects for experiments</td>
		</tr>
		<tr>
			<td>Finnpipette F1&#160;</td>
			<td>ThermoFisher Scientific</td>
			<td>4641080N</td>
			<td>micropipette for dispensing liquids</td>
		</tr>
		<tr>
			<td>Finntip 250 pipette tips</td>
			<td>ThermoFisher Scientific</td>
			<td>9400250</td>
			<td>micropipette tips</td>
		</tr>
		<tr>
			<td>Microscope Glass cover slides (=coverslips) (24 x 24 mm)</td>
			<td>AmScope</td>
			<td>CS-S24-100</td>
			<td>coverslips for viewing the insect&#39;s crop on confocal microscope</td>
		</tr>
	</tbody>
</table>

the ingestion of fluorescent, magnetic nanoparticles for determining fluid-uptake abilities in insects

流体食性昆虫を摂取する環境では、プール、映画、または小さな毛穴に閉じ込められた液体の様々 な。口器の構造と機能の関係を評価する液体の獲得の研究が必要とただし、流体の吸収機構の解明は歴史的構造のアーキテクチャは、実験的証拠と時々 一人旅の観察から推論されます。蝶 (鱗翅目) とハエ (双翅目) の液体の少量を使用して流体吸収能力を評価する手法を報告する.昆虫は、特定の細孔径のフィルター ペーパーから蛍光、磁性ナノ粒子と混合 20% ショ糖液が供給されます。作物 (流体を格納するために使用される内部構造) は、昆虫から削除され、共焦点顕微鏡上に配置。磁石は昆虫が水分を摂取することができるかどうかを示すナノ粒子の存在を確認するための作物で手を振った。この手法を使用して、機構の解明、広範な供給 (毛細管と液体ブリッジの形成) 多孔性の表面からを供給するとき可能性のある鱗翅目、双翅目の間で共有されます。さらに、このメソッドは液餌昆虫、病気の感染、バイオミメティックス、ナノまたはマイクロ導管を含む可能性のある他の研究で重要なを含むさまざまな機能の研究使用できます、液体の輸送には、検証が必要です。

液餌昆虫の中で流体吸収能力のパターンの研究には、餌が発生したときの決意が必要です。ここで説明したプロトコルを使用して、鱗翅目、双翅目の13の間制限の細孔サイズ仮説をテストしています。制限の細孔サイズ仮説は、細孔径が供給管12の直径より小さい場合は、液体充填孔から液餌昆虫を与えることができない状態します...

Watch this Scientific Journal Video about The Ingestion of Fluorescent, Magnetic Nanoparticles for Determining Fluid-uptake Abilities in Insects at JoVE.com

The Ingestion of Fluorescent, Magnetic Nanoparticles for Determining Fluid-uptake Abilities in Insects

流体食性昆虫は、多孔性の表面の液体の微量を取得する能力を持っています。このプロトコルでは、直接蛍光、磁性ナノ粒子供給溶液を用いた多孔質表面から液体を摂取する昆虫の能力を決定する方法について説明します。

蛍光灯、摂取昆虫における流体吸収能力を決定するための磁気ナノ粒子

Fluid-feeding insects ingest a variety of liquids, which are present in the environment as pools, films, or confined to small pores. Studies of liquid ...

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Biology

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の蛍光標識ショウジョウバエハートの構造

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Magnetotactic bacteria comprise a diverse group of aquatic microorganisms that are able to orientate themselves along geomagnetic fields. This behavior is ...

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Simultaneous Measurement of Mitochondrial Calcium and Mitochondrial Membrane Potential in Live Cells by Fluorescent Microscopy

蛍光顕微鏡による生細胞内のミトコンドリアのカルシウムとミトコンドリア膜電位の同時測定

Apart from their essential role in generating ATP, mitochondria also act as local calcium (Ca2+) buffers to tightly regulate intracellular Ca2+ ...

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Here, we describe a microwave synthesis to obtain iron oxide nanoparticles core-doped with 68Ga. Microwave technology enables fast and reproducible ...

High-Throughput Assays of Critical Thermal Limits in Insects

昆虫の臨界熱限界のハイスループットアッセイ

Upper and lower thermal limits of plants and animals are important predictors of their performance, survival, and geographic distributions, and are ...