著者に感謝 Pi 芬、林、氏魏肺・ ツェン、ミス プリンスエド ワード島玄林博士甲陸チュアン フィールド サンプリングとラボ分析と「CATNET」の開発・実用化のための支援乗組員とボード上の調査船海洋研究技術者の支援-I およびサンプリング遠征中に海洋研究 II は大歓迎します。この作品は台湾省科学によって部分的サポートされて、技術付与 104-2611-M-002-019, 97-3114-M-002-006 96-2611-M-002-004 95-2611-M-002-009 91-2611-M-002-007。この原稿は、温輝リー嬢の彼女の巨大な献身と台湾の海洋の研究への貢献のためのメモリに書き込まれます。

次のプロトコルには、有害な化学物質の使用が含まれます。ください安全性データ シート (SDS) を熟読し、制度化学物質の安全性ガイドラインに従ってください。
1. 多層重力ろ過サンプラー準備
<ol><li>
クリーニングのサンプラー
	<ol>
<li>アニオン性プロテアーゼ酵素洗剤の 1% (w/v) と、チューブとろ過の単位を記入し、浸し 24 h. フラッシュ逆浸透膜二重蒸留水 (RO DDW) 多層重力ろ過サンプラー徹底的に 0.1% (v/v) でそれを埋める塩酸 (HCl、試薬グレード)、72 時間浸漬。</li>
		<li>徹底的に逆浸透ダブル蒸留純水 (RO-DD-DIW) 3 〜 5 倍 20 リットル以上の時間とビニール袋に群集を格納と多層重力ろ過サンプラーをフラッシュします。</li>
	</ol>
</li>
	<li>
粒子サンプル コンテナー洗浄/準備
	<ol>
<li>粒子のコンテナーとして低密度ポリエチレン (LDPE、125 mL) またはフッ素化エチレン プロピレン (FEP、125 mL) ボトルを使用します。それぞれ最初アルカリ性洗剤 (マイクロ、1%) の 50% (v/v) 硝酸 (HNO3、試薬グレード)、それから少なくとも 24、48、24 h の 10% (v/v) HCl ソリューションでそれらを浸すことによってボトルをきれい。脱イオン水 (RO-DD-DIW) 2 つの浸漬手順間でボトルをすすいでください。</li>
		<li>最終的な HCl 浸漬後、徹底的に脱イオン水 (RO DD DIW) でボトルを洗浄、クリーン ルーム、クラス 100 のクリーン ベンチの中にボトルを乾燥します。 多層重力ろ過サンプラ洗浄瓶を添付またはきれいにシールは輸送のためそれらを PE ジッパー袋、二重袋のボトルします。</li>
	</ol>
</li>
	<li>
多層重力ろ過サンプラーの群集
	<ol>
<li>サンプラーの上に 6 方向入口に (0.635 cm 外径) 6 4 m 長い化学的に抵抗力がある熱可塑性エラストマー チューブを接続します。</li>
		<li>低密度ポリエチレン サンプル容器 (LDPE 125 mL) を 3 つの異なるメッシュ ナイロン フィルターをクリーン ルーム外に配置されている 10 μ m のメッシュ フィルターをクリーンアップです (下記参照) 後、(ベンチ) の順序で組み立てる 63 μ m メッシュ フィルターで、中高 153 μ m メッシュの内側にフィルターを実行します。輸送、ポリエチレン (PE) 袋の 2 つの層の多層重力ろ過サンプラーを保存し、ポリプロピレン (PP) 出荷用コンテナーに配置。</li>
	</ol>
</li>
</ol>2. サンプリング
<ol><li>
サンプル コレクション
	<ol>
<li>サンプリング サイトに到着すると、調査船の甲板上のコンテナーから多層重力ろ過サンプラーを外し、サンプラーと袋を開ける人があります。その後、PE 手袋を入れて、高いマルチ ボトル サンプリング アレイ上 6 20 L PTFE 被覆サンプリング ボトルの水栓に 6 の 4 m 熱可塑性エラストマー チューブを接続し、このろ過装置に海水を導くことがあります。海水を通過方向の入り江と、粒子/プランクトンが優しく分離/分別ネットを通じて、網の基地で保護される LDPE ボトル 125 mL に落ち着きます。</li>
		<li>海水には (沿岸海水用通常 120 L と外洋水 240 L) が流れて後の順序で各ネットを削除 (まず、153 μ m、63 μ m と 10 μ m 最後に) クラス 100 のクリーン ベンチで、微量金属きれい 0.4 μ m f ネットをスプレーiltered 海水洗浄すべてのプランクトン ネットの内側表面に付着します。125 mL ペットボトルで高濃度粒子/プランクトンを海水を収集します。</li>
		<li>ネットからこれらのボトルを緩めてフィルター酸洗浄吸引ろ過装置を通して再び集中粒子/プランクトンとソリューションの事前の重量を量った、酸洗浄 47 mm の低真空下で 10 μ m 孔サイズのポリカーボネート フィルター条件 (< 5 kPa)。</li>
		<li>その後、サンプラーを流れる 5 L PE 容器内の水の 2 〜 5 L を収集し、酸洗浄吸引ろ過装置でこれらのサンプル水をフィルター海水の少なくとも 20 L を待つ 10 μ m より小さい粒子/プランクトンを収集するにはあらかじめ重量を量られた、酸洗浄、47 mm、0.4 μ m 孔サイズのポリカーボネート フィルター。</li>
		<li>真空ろ過後高純度 DDW 水海水、海の塩の粒子/プランクトンの乾燥重量の決定に及ぼす影響を最小限に抑えることの残基を削除するサンプル フィルターを直ちに洗浄します。壊れやすいプランクトンを損傷を防ぐためにミリリットルのカップルだけにすすぎのボリュームを維持します。</li>
		<li>、この洗浄のステップ後慎重に削除しフィルター真空ろ過装置から、酸洗浄、あらかじめ重量を量られたアクリル プラスチック シャーレ、にサンプル フィルターを格納、ジッパーのついたビニール袋に密封します。さらにサンプル前処理および化学分析のための土地ベースの研究室に戻るまで-20 ° C のフリーザー搭載で袋をしてください。</li>
	</ol>
</li>
</ol>3. サンプル治療
<ol><li>
凍結乾燥と粒子の消化
	<ol>
<li>凍結乾燥機のコレクターの部屋で粒子のサンプル フィルターを置き、マシンをオンにします。機械温度-40 ° C に達すると、機械の真空ポンプをオンに、凍結乾燥プロセスを開始します。 注: 真空度は、0.12 mBar 以下着実に維持する必要があります。くださいユーザー マニュアルを注意深く読み、各ステップの製造元のガイドラインに従ってください。</li>
		<li>72 時間後をオフに凍結乾燥機、乾燥フィルターを削除し、それらの重量を量る。あらかじめ重量を量られたパーフルオロアルコキシ アルカン (PFA) 容器 (容量 60 mL) に乾燥のサンプル フィルターを配置し、船2,3,6,7に集中している超高純度硝酸 3 mL を加えます。</li>
		<li>2.5 kg ・ m、定トルクをトルクレンチで血管を強化し、最初の消化シーケンスの 12 h 130 ° C で従来のオーブンで血管を配置します。冷却した後、血管をオーブンから取り出し、船、船2,3,6,7に超高純度フッ化水素酸 2 mL を追加します。</li>
		<li>2.5 kg ・ m のトルクで血管を強化し、2 番目の消化シーケンスである 12 h 130 ° C で従来のオーブンで血管を配置します。冷却した後、船、船2,3,6,7に 4.5% 純度ホウ酸液 16 mL を追加します。</li>
		<li>2.5 キロ メートルの一定トルクの血管を締め、最終消化順序の 12 h 130 ° C のオーブンでサンプルを消化します。冷却した後、各船の重さし、最終的な質量と各消化ソリューション最終 digestant ボリュームを生成するための具体的な重量を決定します。 注: 特定の質量は、digestant のまさに 1.00 mL の重量を測定によって決定されます。</li>
		<li>慎重に、digestant をさらに微量金属分析用 30 mL 酸洗浄 PE ボトルに注ぐ。</li>
	</ol>
</li>
	<li>
微量金属分析
	<ol>
<li>微量金属元素濃度 (Cd、Cu、Fe、Mn、Ni、および Al) 消化ソリューションの黒鉛炉原子吸光分析計 (吸)6を使用して粒子を決定します。</li>
		<li>精度のテストとして、カナダ国立研究評議会の国立標準研究所の技術から河口域底質標準物質から海底堆積物試料などの標準物質 (CRM) を使用して、米国および欧州委員会の科学と知識のサービスからプランクトン参考資料。プロセスは、CRM、微量金属の認定値の 95% に 107% 回復を与えます。</li>
	</ol>
</li>
</ol>

<ol>
	<li>Jeandel, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=What+did+we+learn+about+ocean+particle+dynamics+in+the+GEOSECS-JGOFS+era.">What did we learn about ocean particle dynamics in the GEOSECS-JGOFS era.</a> Progr. Oceanogr. 133, 6-16 (2015).</li><li>Lam, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+for+analyzing+the+concentration+and+speciation+of+major+and+trace+elements+in+marine+particles.">Methods for analyzing the concentration and speciation of major and trace elements in marine particles.</a> Progr. Oceanogr. 133, 32-42 (2015).</li><li>Collier, R., Edmond, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+trace+element+geochemistry+of+marine+biogenic+particulate+matter.">The trace element geochemistry of marine biogenic particulate matter.</a> Progr. Oceanogr. 13, 113-199 (1984).</li><li>Donat, J. R., Bruland, K. W., Steinnes, E., Salbu, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trace+elements+in+the+oceans.">Trace elements in the oceans.</a> Trace Elements in Natural Waters. , 247-280 (1995).</li><li>Wen, L. -. S., Santschi, P., Tang, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interaction+between+radioactively+labeled+colloids+and+natural+particles:+evidence+for+colloidal+pumping.">Interaction between radioactively labeled colloids and natural particles: evidence for colloidal pumping.</a> Geochim. Cosmochim. Ac. 61, 2867-2878 (1997).</li><li>Wen, L. -. S., Warnken, K., Santschi, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+organic+carbon,+iron,+and+aluminium+oxyhydroxides+as+trace+metal+carriers:+Comparison+between+the+Trinity+River+and+the+Trinity+River+Estuary+(Galveston+Bay,+Texas).">The role of organic carbon, iron, and aluminium oxyhydroxides as trace metal carriers: Comparison between the Trinity River and the Trinity River Estuary (Galveston Bay, Texas).</a> Mar. Chem. 112, 20-37 (2008).</li><li>Hurd, D., Spencer, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Marine+particles:+analysis+and+characterization.">Marine particles: analysis and characterization.</a> American Geophysical Union. , (1991).</li><li>Martin, J. H., Knauer, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+elemental+composition+of+plankton.">The elemental composition of plankton.</a> Geochim. Cosmochim. Ac. 37, 1639-1653 (1973).</li><li>Morel, F., Price, N. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+biogeochemical+cycles+of+trace+metals+in+the+oceans.">The biogeochemical cycles of trace metals in the oceans.</a> Science. 300, 944-947 (2003).</li><li>Ho, T. -. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+elemental+composition+of+some+marine+phytoplankton.">The elemental composition of some marine phytoplankton.</a> J. Phycol. 39, 1145-1159 (2003).</li><li>McDonnell, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+oceanographic+toolbox+for+the+collection+of+sinking+and+suspended+marine+particles.">The oceanographic toolbox for the collection of sinking and suspended marine particles.</a> Prog. Oceanogr. 133, 17-31 (2015).</li><li>Wen, L. -. S., Li, W. -. H., Zhuang, G. -. Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Multiple layer filtering and collecting device. , (2005).</li><li>Ho, T. -. Y., Wen, L. -. S., You, C. -. F., Lee, D. -. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+trace-metal+composition+of+size+fractionated+plankton+in+the+South+China+Sea:+biotic+versus+abiotic+sources.">The trace-metal composition of size fractionated plankton in the South China Sea: biotic versus abiotic sources.</a> Limnol. Oceanogr. 52, 1776-1788 (2007).</li><li>Hsu, R., Liu, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In-situ+estimations+of+the+density+and+porosity+of+flocs+of+varying+sizes+in+a+submarine+canyon.">In-situ estimations of the density and porosity of flocs of varying sizes in a submarine canyon.</a> Mar. Geol. 276, 105-109 (2010).</li><li>Liao, W. -. H., Yang, S. -. C., Ho, T. -. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trace+metal+composition+of+size-fractionated+plankton+in+the+Western+Philippine+Sea:+the+impact+of+anthropogenic+aerosol+deposition.">Trace metal composition of size-fractionated plankton in the Western Philippine Sea: the impact of anthropogenic aerosol deposition.</a> Limnol Oceanogr. , (2017).</li><li>Grasshoff, K., Kremling, K., Ehrhardt, 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> Methods of seawater analysis. , (2007).</li></ol>

共同著者氏アラン荘は特許の共同所有者と興味を持つユーザーのコレクション装置を製造 (中国楽器株式) 会社のゼネラル マネージャー。特許は、9th、2015年125 月に終わった。

中に検体の採取、処理、前処理、分析の目的で細心の注意が必要です取得の信頼性の高いトレース プランクトンと一般的に非常に低濃度で存在している、天然水中の懸濁粒子の金属濃度汚染を低減します。そのため、設計およびサンプリング装置、サンプル容器、材料を準備する手順を収集する、プロセスのサンプルが海洋環境中の微量金属の高品質データ取得に向けたすべての重要な手順...

海洋中の粒子は、海洋物質循環1で重要な役割を果たします。サイズ、鉱物組成などの粒子の特性の最も深く 1 つの地質や前浜の設定から別の2変更できます。加えて、海洋における元素の分布が海洋植物プランクトンのライフ サイクルに関連付けられている: 成長、死、沈没、赤潮3,4。海洋の粒子にまたがるサブミクロン粒子から大きな凝集体に至るまでのサイズの少なくとも 4 桁 (> 5 mm)。ほとんどの粒子は、ウイルスの換散、滲出、分泌、糞便のペレット製造などのプロセスから派生生物学的。その他の粒子は、細胞、細胞の残骸や隆起材料1の物理的な凝固から形成されます。両方の物質循環と生物学的現象と粒子4,5,6内で粒子の様々 な化学的および生物学的特性を制御します。これらの粒子は動物プランクトンや saprotrophs など、いくつかの有機体のための食糧源と同様、重要な生息地です。したがって、粒子の運命は頻繁に生物学的過程と粒子の周りによって変更することができます彼らのサイズに関連します。
海洋の粒子を通常サンプリング、濾過がこのアプローチは海洋の粒子の構成内容と同種ではないので、粒子の特性を識別するに特定のあいまいさを紹介します。主体がほぼ完全に懸濁液、小型・低密度粒子の懸濁粒子が懸濁液のより大きくより密度の高い粒子の変化量と流体条件によっては、時間の短い期間にのみ混在します。7. プランクトン試料の微量金属の組成の最初のレポートは研究容器8プランクトン トウまたは中断するプランクトン ネットで収集されました。著者はしばしば金属粒子を発見し、ペイント チップ サンプルの化学分析のための海洋の粒子のサンプリング時の汚染の深刻な問題を示唆しています。その他、純ゴムいかだによって曳航または塩化ビニル (PVC) を使用して-手のウィンチ3。粒子の信頼性の高いサンプリングの難しさ、特に微量元素より困難海洋粒子の化学組成の私達の理解の進歩になります。そのため、植物プランクトン中の微量元素濃度に関する最も重要な情報は、文化研究9,10から来ています。このような認識、海洋科学者が過去 30 年間11上海で粒子を研究するための新しいメソッドを作成する動機となった。
海洋学者は、その場でろ過、船上濾過など様々 なサンプリング技術を使用しているし、セジメント トラップ11。特に外洋と深海粒子濃度が非常に低いであるため、非汚染のサンプルを収集する海水の大量の処理に挑戦することができます (0.001 〜 0.1 mg/L)。大量の海水微量金属元素濃度を測定するパーティクルの十分な量を取得するフィルターを適用する必要です。一部の研究者は、沈降粒子から粒子を分離するのにサイズ分画測定法を使用しています。しかし、粒径、空隙率、密度、および図形は、すべて影響粒子沈降速度をことができます。セジメント トラップは、それらが沈降粒子設計されているため、懸濁粒子を収集するために実用的なツールではありません。したがって、最小の汚染懸濁粒子の十分な量を集めることができるサンプリング ・治療法を開発することが重要です。したがって、その場で濾過により分画のサイズはまだ海洋のサンプリング ツールボックスで有望なツールそれは海洋の粒子のダイナミクスに関する重要な情報を明らかにすることができますので。ここでは、正常にテスト述べる微量金属をきれい、ポリテトラフルオロ エチレン (PTFE) から 1 つのパスでボード上の海水のサンプリング装置、大容量 (120-240 L) を扱うことができる多層重力ろ過コーティングで水のサンプリング ボトル、マルチ ボトル サンプリング配列。この捕集装置が順番に酸洗浄した合成ナイロン ネットを使用し、ネットが優しくサイズ分画された懸濁物質と植物プランクトン12,13,を収集するポリカーボネート容器に含まれている14,15 (図 1)。この作業の目的は、金属粒子の関連付けと海洋環境における反応ダイナミクスを研究するためより良いツールを提供し、さまざまなプランクトン、粒子、およびこれらの微量金属の運命の私達の理解を改善環境。

<table><tbody><tr><td>thermoplastic elastomer (C-Flex) Tubings</td><td>Cole Palmer</td><td>EW-06424-67</td><td>O.D. 0.635 cm, Opaque White 1/8&quot;ID x 1/4&quot;OD, 25 ft/pack</td></tr><tr><td>LDPE Bottle (Nalgene)</td><td>ThermoFisher Scientific</td><td>2103-0004</td><td>125 mL, Nalgene Wide-Mouth LDPE Bottles with Closure</td></tr><tr><td>anionic protease enzyme detergent detergent (Tergazyme)</td><td>Alconox</td><td>1104-1</td><td>1&amp;times;4 lb box (1.8 kg)</td></tr><tr><td>Hydrochloric Acid</td><td>Sigma-Aldrich</td><td>258148</td><td>Reagent grade</td></tr><tr><td>Nitric acid</td><td>Sigma-Aldrich</td><td>695025</td><td>Reagent grade</td></tr><tr><td>alkaline detergnet (Micro)</td><td>Cole Palmer</td><td>EW-99999-14</td><td>Micro-90 Cleaning Solution</td></tr><tr><td>polycarbonate filter, 47 mm, 0.4 &amp;micro;m</td><td>Sigma-Aldrich</td><td>WHA111107</td><td>Whatman Nuclepore Track-Etched Membranes, diam. 47 mm, pore size 0.4 &amp;mu;m, polycarbonate</td></tr><tr><td>polycarbonate filter, 47 mm, 10 &amp;micro;m</td><td>Sigma-Aldrich</td><td>WHA111115</td><td>Whatman Nuclepore Track-Etched Membranes, diam. 47 mm, pore size 10 &amp;mu;m, polycarbonate</td></tr><tr><td>PFA vessel, 60 ml capacity</td><td>Savillex</td><td>300-060-03</td><td>60 mL Digestion Vessel, Flat Interior, Flat Exterior, Buttress Threaded Top</td></tr><tr><td>Nitric acid, ultrapure</td><td>Seastar Chemicals</td><td>N/A</td><td>BASELINE Nitric Acid</td></tr><tr><td>HF, ultrapure</td><td>Seastar Chemicals</td><td>N/A</td><td>BASELINE Hydrofluoric Acid</td></tr><tr><td>Boric acid, ultrapure</td><td>Seastar Chemicals</td><td>N/A</td><td>BASELINE Hydrobromic Acid</td></tr><tr><td>polyethylene (PE) gloves</td><td>Safty Zone</td><td>GDPL-MD-5</td><td>Clear Powder Free Polyethylene Gloves</td></tr><tr><td>Multiple layer filtering and collecting device</td><td>Sino Instrumnets Co. Ltd</td><td>not available</td><td>Multiple layer filtering and collecting device, CATNET</td></tr><tr><td>10 um Nylon filters, Nitex</td><td>Dynamic Aqua-Supply Ltd.</td><td>NTX 10</td><td>Nitex - Standard Widths (40 - 44 inches)</td></tr><tr><td>60 um Nylon filters, Nitex</td><td>Dynamic Aqua-Supply Ltd.</td><td>NTX 60</td><td>Nitex - Standard Widths (40 - 44 inches)</td></tr><tr><td>150 um Nylon filters, Nitex</td><td>Dynamic Aqua-Supply Ltd.</td><td>NTX 150</td><td>Nitex - Standard Widths (40 - 44 inches)</td></tr><tr><td>torque wrench</td><td>Halfords</td><td>200238</td><td>Halfords Professional Torque Wrench 8-60Nm</td></tr><tr><td>multi-bottle sampling array, Rosette</td><td>General Oceanics</td><td>Model 1018</td><td>Rosette Sampler</td></tr><tr><td>PTFE-coated sampling bottles, GO-Flo</td><td>General Oceanics</td><td>108020T</td><td>GO-Flo water sampler teflon coated</td></tr><tr><td>Marine sediment reference materials</td><td>National Research Council Canada</td><td>MESS-3</td><td></td></tr><tr><td>Estuarine sediment standard reference material</td><td>National Institute of Standards and Technology</td><td>1646a</td><td></td></tr><tr><td>Plankton reference material</td><td>The European Commission&#039;s science and knowledge service</td><td>CRM414</td><td></td></tr></tbody></table>

an ultra-clean multilayer apparatus for collecting size fractionated marine plankton and suspended particles

海で多くの微量元素の分布、成長、死、および海洋プランクトンの赤潮と中断された沈降粒子に強く関連付けられています。すべてプラスチック (ポリプロピレン、ポリカーボネート) 多層ろ過システム海で浮遊粒子状物質 (SPM) のコレクションを紹介します。この超クリーン サンプリング デバイスを設計されており、微量元素の研究用に開発されました。すべての非金属材料の細心の選択、プロシージャ フローをインラインでの使用率は、サンプリング中に可能な金属汚染を最小限に抑えます。このシステムは、正常にテストされており、沿岸や外洋海域でさまざまな大きさの粒子における微量元素 (例えばFe、Al、Mn、Cd、Cu、Ni) を決定するために調整します。サウス イースト アジア時系列 (席) 駅南シナ海からの結果は、日内変動と有光層におけるプランクトンの分布することができます簡単に解決し、認識ことを示します。台湾海峡の海面でサイズ分画された粒子の化学分析は示唆大きい粒子 (> 153 μ m) 小さい粒子 (10 63 μ m) が主無機物質から成る間、得られた大抵生物学的。Cd から離れて増加するサイズの金属 (Fe、Al、Mn, Cu, Ni) の濃度が減少しました。

近代海洋学の発展に伴い、それは「クリーン技術」を使用する一般的な練習では今海洋粒子やプランクトンの金属濃度の正確なトレースを取得します。天然水の粒子の大半は μ g/L の範囲に低 mg/l なので海水の大量の治療は周囲の環境で様々 な粒子の微量金属元素の地球化学的および生物学的効果を検討する必要。きれいな、多層重力ろ過 (「CATNET」) のサンプリング...

Watch this Scientific Journal Video about An Ultra-clean Multilayer Apparatus for Collecting Size Fractionated Marine Plankton and Suspended Particles at JoVE.com

An Ultra-clean Multilayer Apparatus for Collecting Size Fractionated Marine Plankton and Suspended Particles

サイズ分画した海洋プランクトンや懸濁粒子を収集するための超きれいな積層装置

プランクトンや懸濁粒子は、海の物質循環に大きな役割を果たします。ここでは、海水の大量の処理能力を持つ、超クリーン、低応力法の粒子と海でプランクトンの様々 なサイズのコレクションを提供します。

The distributions of many trace elements in the ocean are strongly associated with the growth, death, and re-mineralization of marine plankton and those ...

an-ultra-clean-multilayer-apparatus-for-collecting-size-fractionated

Universidad de Cartagena UDEC

Research

JoVE Journal

Environment

8.8K ビュー.  National Taiwan University. プランクトンや懸濁粒子は、海の物質循環に大きな役割を果たします。ここでは、海水の大量の処理能力を持つ、超クリーン、低応力法の粒子と海でプランクトンの様々 なサイズのコレクションを提供します。

ビデオ: An Ultra-clean Multilayer Apparatus for Collecting Size Fractionated Marine Plankton and Suspended Particles

Unraveling the Unseen Players in the Ocean - A Field Guide to Water Chemistry and Marine Microbiology

Here we introduce a series of thoroughly tested and well standardized research protocols adapted for use in remote marine environments. The sampling protocols include the assessment of resources available to the microbial community (dissolved organic carbon, particulate organic matter, inorganic nutrients), and a comprehensive description of the viral and bacterial communities (via direct viral and microbial counts, enumeration of autofluorescent microbes, and construction of viral and microbial metagenomes). We use a combination of methods, which represent a dispersed field of scientific disciplines comprising already established protocols and some of the most recent techniques developed. Especially metagenomic sequencing techniques used for viral and bacterial community characterization, have been established only in recent years, and are thus still subjected to constant improvement. This has led to a variety of sampling and sample processing procedures currently in use. The set of methods presented here provides an up to date approach to collect and process environmental samples. Parameters addressed with these protocols yield the minimum on information essential to characterize and understand the underlying mechanisms of viral and microbial community dynamics. It gives easy to follow guidelines to conduct comprehensive surveys and discusses critical steps and potential caveats pertinent to each technique.

Here, we present a comprehensive protocol to assess the organic and inorganic nutrient availability and the abundance and structure of microbial and viral communities in remote marine environments.

水化学と海洋微生物学へのフィールドガイド - 海における幽玄選手の解明

Here we introduce a series of thoroughly tested and well standardized research protocols adapted for use in remote marine environments. The sampling ...

環境学

VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders

Benthic suspension feeders play essential roles in the functioning of marine ecosystems. By filtering large volumes of water, removing plankton and detritus, and excreting particulate and dissolved compounds, they serve as important agents for benthic-pelagic coupling. Accurately measuring the compounds removed and excreted by suspension feeders (such as sponges, ascidians, polychaetes, bivalves) is crucial for the study of their physiology, metabolism, and feeding ecology, and is fundamental to determine the ecological relevance of the nutrient fluxes mediated by these organisms. However, the assessment of the rate by which suspension feeders process particulate and dissolved compounds in nature is restricted by the limitations of the currently available methodologies. Our goal was to develop a simple, reliable, and non-intrusive method that would allow clean and controlled water sampling from a specific point, such as the excurrent aperture of benthic suspension feeders, in situ. Our method allows simultaneous sampling of inhaled and exhaled water of the studied organism by using minute tubes installed on a custom-built manipulator device and carefully positioned inside the exhalant orifice of the sampled organism. Piercing a septum on the collecting vessel with a syringe needle attached to the distal end of each tube allows the external pressure to slowly force the sampled water into the vessel through the sampling tube. The slow and controlled sampling rate allows integrating the inherent patchiness in the water while ensuring contamination free sampling. We provide recommendations for the most suitable filtering devices, collection vessel, and storing procedures for the analyses of different particulate and dissolved compounds. The VacuSIP system offers a reliable method for the quantification of undisturbed suspension feeder metabolism in natural conditions that is cheap and easy to learn and apply to assess the physiology and functional role of filter feeders in different ecosystems.

VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders

We introduce the VacuSIP, a simple, non-intrusive, and reliable method for clean and accurate point sampling of water. The system was developed and evaluated for the simultaneous collection of the water inhaled and exhaled by benthic suspension feeders in situ, to cleanly measure removal and excretion of particulate and dissolved compounds.

VacuSIP、のための改良INEX方法<em&gt;その場で</em&gt;微粒子と溶解した化合物の測定は、アクティブサスペンションフィーダによって処理します

Benthic suspension feeders play essential roles in the functioning of marine ecosystems. By filtering large volumes of water, removing plankton and ...

Protocol for Microplastics Sampling on the Sea Surface and Sample Analysis

Microplastic pollution in the marine environment is a scientific topic that has received increasing attention over the last decade. The majority of scientific publications address microplastic pollution of the sea surface. The protocol below describes the methodology for sampling, sample preparation, separation and chemical identification of microplastic particles. A manta net fixed on an &#187;A frame&#171; attached to the side of the vessel was used for sampling. Microplastic particles caught in the cod end of the net were separated from samples by visual identification and use of stereomicroscopes. Particles were analyzed for their size using an image analysis program and for their chemical structure using ATR-FTIR and micro FTIR spectroscopy. The described protocol is in line with recommendations for microplastics monitoring published by the Marine Strategy Framework Directive (MSFD) Technical Subgroup on Marine Litter. This written protocol with video guide will support the work of researchers that deal with microplastics monitoring all over the world.

The protocol below describes the methodology for: microplastics sampling on the sea surface, separation of microplastic and chemical identification of particles. This protocol is in line with the recommendations for microplastics monitoring published by the MSFD Technical Subgroup on Marine Litter.

海面とサンプル分析にMicroplasticsサンプリングのためのプロトコル

Microplastic pollution in the marine environment is a scientific topic that has received increasing attention over the last decade. The majority of ...

Sampling, Sorting, and Characterizing Microplastics in Aquatic Environments with High Suspended Sediment Loads and Large Floating Debris

The ubiquitous presence of plastic debris in the ocean is widely recognized by the public, scientific communities, and government agencies. However, only recently have microplastics in freshwater systems, such as rivers and lakes, been quantified. Microplastic sampling at the surface usually consists of deploying drift nets behind either a stationary or moving boat, which limits the sampling to environments with low levels of suspended sediments and floating or submerged debris. Previous studies that employed drift nets to collect microplastic debris typically used nets with &#8805;300 &#181;m mesh size, allowing plastic debris (particles and fibers) below this size to pass through the net and elude quantification. The protocol detailed here enables: 1) sample collection in environments with high suspended loads and floating or submerged debris and 2) the capture and quantification of microplastic particles and fibers &lt;300 &#181;m. Water samples were collected using a peristaltic pump in low-density polyethylene (PE) containers to be stored before filtering and analysis in the lab. Filtration was done with a custom-made microplastic filtration device containing detachable union joints that housed nylon mesh sieves and mixed cellulose ester membrane filters. Mesh sieves and membrane filters were examined with a stereomicroscope to quantify and separate microplastic particulates and fibers. These materials were then examined using a micro-attenuated total reflectance Fourier transform infrared spectrometer (micro ATR-FTIR) to determine microplastic polymer type. Recovery was measured by spiking samples using blue PE particulates and green nylon fibers; percent recovery was determined to be 100% for particulates and 92% for fibers. This protocol will guide similar studies on microplastics in high velocity rivers with high concentrations of sediment. With simple modifications to the peristaltic pump and filtration device, users can collect and analyze various sample volumes and particulate sizes.

Most microplastic research to date has occurred in marine systems where suspended solid levels are relatively low. Focus is now shifting to freshwater systems, which may feature high sediment loads and floating debris. This protocol addresses collecting and analyzing microplastic samples from aquatic environments that contain high suspended solid loads.

サンプリング、並べ替え、および高い浮遊負荷と大きい浮遊残骸水圏環境における Microplastics の特性

The ubiquitous presence of plastic debris in the ocean is widely recognized by the public, scientific communities, and government agencies. However, only ...

Design and Use of an Apparatus for Quantifying Bivalve Suspension Feeding at Sea

As shellfish aquaculture moves from coastal embayments and estuaries to offshore locations, the need to quantify ecosystem interactions of farmed bivalves (i.e., mussels, oysters, and clams) presents new challenges. Quantitative data on the feeding behavior of suspension-feeding mollusks is necessary to determine important ecosystem interactions of offshore shellfish farms, including their carrying capacity, the competition with the zooplankton community, the availability of trophic resources at different depths, and the deposition to the benthos. The biodeposition method is used to quantify feeding variables in suspension-feeding bivalves in a natural setting and represents a more realistic proxy than laboratory experiments. This method, however, relies upon a stable platform to satisfy the requirements that water flow rates supplied to the shellfish remain constant and the bivalves are undisturbed. A flow-through device and process for using the biodeposition method to quantify the feeding of bivalve mollusks were modified from a land-based format for shipboard use by building a two-dimensional gimbal table around the device. Planimeter data reveal a minimal pitch and yaw of the chambers containing the test shellfish despite boat motion, the flow rates within the chambers remain constant, and operators are able to collect the biodeposits (feces and pseudofeces) with sufficient consistency to obtain accurate measurements of the bivalve clearance, filtration, selection, ingestion, rejection, and absorption at offshore shellfish aquaculture sites.

A flow-through device for using the biodeposition method to quantify filtration and feeding behavior of bivalve mollusks was modified for shipboard use. A two-dimensional gimbal table built around the device isolates the apparatus from boat motion, thereby allowing the accurate quantification of bivalve filtration variables at offshore shellfish aquaculture sites.

デザインと二枚貝のサスペンションが海で餌を定量化するための装置の使用

As shellfish aquaculture moves from coastal embayments and estuaries to offshore locations, the need to quantify ecosystem interactions of farmed bivalves ...

Large Volume (20L+) Filtration of Coastal Seawater Samples

The workflow begins with the collection of coastal marine waters for downstream microbial community, nutrient and trace gas analyses. For this method, samples were collected from the deck of the HMS John Strickland operating in Saanich Inlet. This video documents large volume (&ge;20 L) filtration of microbial biomass, ranging between 0.22&mu;m and 2.7&mu;m in diameter, from the water column. Two 20L samples can be filtered simultaneously using a single pump unit equipped with four rotating heads. Filtration is done in the field on extended trips, or immediately upon return for day trips. It is important to record the amount of water passing through each sterivex filter unit. To prevent biofilm formation between sampling trips, all filtration equipment must be rinsed with dilute HCl and deionized water and autoclaved immediately after use. This procedure will take approximately 5 hours plus an additional hour for clean up.

Large Volume 20L+ Filtration of Coastal Seawater Samples

This video documents large volume (&ge;20 L) filtration of microbial biomass, ranging between 0.22&mu;m and 2.7&mu;m in diameter, from the water column.

沿岸海水サンプルの大容量（20L +）ろ過

The workflow begins with the collection of coastal marine waters for downstream microbial community, nutrient and trace gas analyses. For this method, ...

生物学

Small Volume (1-3L) Filtration of Coastal Seawater Samples

The workflow begins with the collection of coastal marine waters for downstream microbial community, nutrient and trace gas analyses. For today s demonstration samples were collected from the deck of the HMS John Strickland operating in Saanich Inlet. This video documents small volume (~1 L) filtration of microbial biomass from the water column. The protocol is an extension of the large volume sampling protocol described earlier, with one major difference: here, there is no pre-filtration step, so all size classes of biomass are collected down to the 0.22 &mu;m filter cut-off. Samples collected this way are ideal for nucleic acid analysis. The set-up, filtration, and clean-up steps each take about 20-30 minutes. If using two peristaltic pumps simultaneously, up to 8 samples may be filtered at the same time. To prevent biofilm formation between sampling trips, all filtration equipment must be rinsed with dilute HCl and deionized water and autoclaved immediately after use.

Small Volume 1-3L Filtration of Coastal Seawater Samples

This video documents small volume (~1 L) filtration of microbial biomass from the water column.

沿岸海水試料の少量（1 - 3L）のろ過

The workflow begins with the collection of coastal marine waters for downstream microbial community, nutrient and trace gas analyses. For today s ...

A Microfluidic Platform for Precision Small-volume Sample Processing and Its Use to Size Separate Biological Particles with an Acoustic Microdevice

A major advantage of microfluidic devices is the ability to manipulate small sample volumes, thus reducing reagent waste and preserving precious sample. However, to achieve robust sample manipulation it is necessary to address device integration with the macroscale environment. To realize repeatable, sensitive particle separation with microfluidic devices, this protocol presents a complete automated and integrated microfluidic platform that enables precise processing of 0.15&#8211;1.5 ml samples using microfluidic devices. Important aspects of this system include modular device layout and robust fixtures resulting in reliable and flexible world to chip connections, and fully-automated fluid handling which accomplishes closed-loop sample collection, system cleaning and priming steps to ensure repeatable operation. Different microfluidic devices can be used interchangeably with this architecture. Here we incorporate an acoustofluidic device, detail its characterization, performance optimization, and demonstrate its use for size-separation of biological samples. By using real-time feedback during separation experiments, sample collection is optimized to conserve and concentrate sample. Although requiring the integration of multiple pieces of equipment, advantages of this architecture include the ability to process unknown samples with no additional system optimization, ease of device replacement, and precise, robust sample processing.

This protocol describes a system architecture for performing automated small volume (0.15&#8211;1.5 ml) particle separations using a microfluidic device, and discusses methods to optimize acoustofluidic device performance and operation.

マイクロ流体精密小容量サンプル処理のためのプラットフォームとアコースティックマイクロデバイスで区切り生物学的粒子サイズのためのその使用

A major advantage of microfluidic devices is the ability to manipulate small sample volumes, thus reducing reagent waste and preserving precious sample. ...

生物工学

Separation and Identification of Conventional Microplastics from Farmland Soils

Microplastics (MPs) pollution in the terrestrial environment has received increasing attention over the last decade, with increasing studies describing the numbers and types of MPs in different soil systems and their impacts on soil and crop health. However, different MPs extraction and analytical methods are used, limiting opportunities to compare results and generate reliable evidence for industry advice and policymakers. Here, we present a protocol that describes the methodology for sampling, separation, and chemical identification of conventional MPs&#160;from soil. The method is low-cost, and the materials are readily available. This enhances operational ease and may help with widespread adoption. The protocol provides detailed information on sample collection from the top 0-30 cm of soil using plastic-free utensils; simulation of different soil types through the use of various solid media (such as bentonite clay, silicon dioxide, and non-contaminated soil), with the addition of the same mass&#160;of polyethylene(PE)-MPs for subsequent quantification; density separation of plastic particles utilizing saturated sodium chloride (NaCl) solution and digestion of organic impurities in the supernatant using 4 M sodium hydroxide (NaOH) solution; quantification of particles using fluorescent microscopy after Nile Red staining; and polymer identification using micro&#160;Fourier-Transform Infrared Spectroscopy (&#956;-FTIR) or Laser-Direct Infrared (LDIR) spectroscopy. The MPs recovery rate ranged from 83% - 90% for the abovementioned media. This protocol presents an efficient method for soil MPs analysis that is optimized for feasibility, applicability, and cost-effectiveness. Moreover, the video accompanied can guide the process of analyzing the soil MPs step-by-step virtually. This study is dedicated to standardizing the methods for soil MPs analysis, enhancing the connectivity and comparability of measurements, and establishing a foundation for more standardized and scientific research.

Presented here is a method for extracting microplastics from soil and identifying their polymer types. The method has been optimized for execution, applicability, and cost-effectiveness. It lays a scientific foundation for standardizing the analytical method to identify microplastics in soils.

農地土壌からの従来のマイクロプラスチックの分離と同定

Microplastics (MPs) pollution in the terrestrial environment has received increasing attention over the last decade, with increasing studies describing ...

ヒト尿からの連続的な細胞外小胞分離のためのマイクロ流体デバイス

Single Step Isolation of Extracellular Vesicles from Large-Volume Samples with a Bifurcated A4F Microfluidic Device

Extracellular vesicles (EVs) hold immense potential for various biomedical applications, including diagnostics, drug delivery, and regenerative medicine. Nevertheless, the current methodologies for isolating EVs present significant challenges, such as complexity, time consumption, and the need for bulky equipment, which hinders their clinical translation. To address these limitations, we aimed to develop an innovative microfluidic system based on cyclic olefin copolymer-off-stoichiometry thiol-ene (COC-OSTE) for the efficient isolation of EVs from large-volume samples in a continuous manner. By utilizing size and buoyancy-based separation, the technology used in this study achieved a significantly narrower size distribution compared to existing approaches from urine and cell media samples, enabling the targeting of specific EV size fractions in future applications. Our innovative COC-OSTE microfluidic device design, utilizing bifurcated asymmetric flow field-flow fractionation technology, offers a straightforward and continuous EV isolation approach for large-volume samples. Furthermore, the potential for mass manufacturing of this microfluidic device offers scalability and consistency, making it feasible to integrate EV isolation into routine clinical diagnostics and industrial processes, where high consistency and throughput are essential requirements.

著者スポットライト:バイオリアクター統合のための非対称フィールドフローフラクショネーション

Extracellular vesicles hold immense promise for biomedical applications, but current isolation methods are time-consuming and impractical for clinical use. In this study, we present a microfluidic device that enables the direct isolation of extracellular vesicles from large volumes of biofluids in a continuous manner with minimal steps.

二股A4Fマイクロ流体デバイスを用いた大量サンプルからの細胞外小胞のシングルステップ分離(英語)

Extracellular vesicles (EVs) hold immense potential for various biomedical applications, including diagnostics, drug delivery, and regenerative medicine. ...