The authors would like to convey their gratitude for the technical support provided by National Synchrotron Radiation Research Center (NSRRC) (Taiwan). The authors are grateful for the financial support provided by the Ministry of Science and Technology of the Republic of China (Taiwan) and the Industrial Technology Research Institute (Taiwan).

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The authors have nothing to disclose.

双極子支援SPEのマイクロチップの製造のための詳細な手順は、上記に提示しました。このセクションでは、PMMAのC-Clで部分および微量金属イオンを決意するための抽出媒体として使用されたのCl含有PMMAの可能性の移植に関して修正プロトコルの有用性は、ありますステップバイステップを評価しました。表面の検証のために、試料の種類は分析機器との適合性に基づいて選択しました。換言すれば、同様の工程を経て調製された試験サンプルの種類は、分析機器の要件に従って決定しました。例えば、粉末充填型のサンプルは、LA-ICP-MS、ラマン分光に使用したのに対し、基板型サンプルは、接触角の測定に使用し、XANES解析します。 最初は、化学官能アタッシュケースが受ける変化を監視します提案されている手順の間PMMAの表面にD、各工程を行ったに対応する得られた生成物に対する接触角分析（ 図2）。 図2に表示されているように、接触角の変化は明らかに表面変化は、修正手順の間に発生したことが示され、そして最終生成物を測定したところ80.3°±0.43°の接触角は、以前の結果を報告した。21と一致しましたまた、変性PMMAのC-Clでの成分の存在はまた、LA-ICP-MS分析によって確認しました。ネイティブPMMAをアブレーションすることによって得られた結果と比較して、CLには別個の信号はC-Clで部分で修飾されたPMMAをアブレーションすることによってexpectably観察された（ 図3（a）参照 ）。 ラマンスペクトルは、さらに、PMMAのC-Clで部分の結合を検証するために収集しました。 Figuに示すように図3（b）再もしくはCCl 2非対称伸縮振動に関連する2つの特徴的なピーク682 cm -1で718 cm -1で修正されたPMMAのスペクトル中で観察したウィリスらによって報告された結果とのことで適度に良好な一致22、ヘンドラら 23言い換えると、PMMAのC-Clでの部分の結合が正常に変更した後に達成され得ます。 また、本研究で提案された抽出機構を解明するために、XANES解析を使用しました。 図4に示されるように、高度に電気陰性のC-Clで部分と正に帯電した金属イオンとの間の相互作用は、MN 2+イオンで ​​処理された変性PMMAに対応XANESスペクトルにおける優勢な吸収端が存在することによって確認することができました。このように、双極子静電相互作用は確かTRのためのオンチップ抽出に適用されますエースの金属分析します。台湾の二つの川から採取した水サンプルの詳細な分析結果を他の場所に記載されている。16 我々の知る限り、これは微量金属イオンの決意のためのオンチップSPE反応で革新的な作業戦略を利用する最初の試みであり、開発したデバイスは、他のオンチップSPE技術と比較して有意に耐久性であったこと（ すなわち、 、160以上の分析作品）が、抽出効率の点で大幅な劣化なしに達成することができます。そのような抽出機構は、主として高度に電気陰性のC-Clで部分と正に帯電した金属イオンとの間の相互作用に依存しているため、それにもかかわらず、提案された技術は、これまで負に帯電した種を抽出するには不向きであることが予想されました。

環境管理と汚染防止の観点から、深刻な汚染や毒物学的問題を引き起こす微量金属は、世界的に懸念されます。生試料中の予想外の共存化学種は、多くの場合、トレースの量で存在する分析物の正確な決意を妨げるため、適切なオンチップ・サンプル前処理技術が広く、処理の成功への鍵とチップベースのプラットフォームを介して、実際のサンプルを分析することとして受け入れられています。利用可能な技術のうち1、オンチップ固相抽出（SPE）微量金属のために特に人気が同時に実行される試料のクリーンアップおよび検体前濃縮を可能にするこの技術は、複雑な塩マトリックスからの金属イオンの単離のために非常に有用であるので、分析。 2,3 微量金属の決意に使用されるオンチップSPE技術の進歩は着実に進化してきました。初期の頃には、トン彼はチップを樹脂充填SPEユニットを構成するためにマイクロチャネルに市販の樹脂をロードすることによって調製したSPE 4-7これは時折、樹脂保持可能な形態への金属イオンの変換を可能にするように誘導体化される検体を必要とした。4代替方法チップベースのSPEデバイスの製造のための単純な表面改質後の微量金属の回収のためのSPEの吸着剤としてのチップチャネルを利用することである。8近年、磁性ナノ粒子（MNPの）および特定の化学物質の取り込みを含む新興の傾向を見てきました金属イオンを効率的に保持することができる官能基を含有します。商業的な樹脂とは対照的に、のMNPは、それらが外部磁界Tの助けを借りてマイクロチャンネルに充填された後にγメルカプトプロピルトリメトキシシラン（γ-MPTS）9とアミノベンジルエチレンジアミン四酢酸（ABEDTA）10のような化合物で修飾されていますO金属イオンの選択的抽出を実現します。 オンチップSPE技術の開発における重要な進歩を目撃してきたが、報告された技術は、典型的には、イオン交換またはキレート化のいずれかに基づいて機能します。このような技術の使用は、分析性能を維持するためにエアコン、洗濯、又は再生に関連するものを含む不可避操作手順を必要とするという欠点があります。残念ながら、追加の操作手順の必要性は、各分析に必要な時間を延長するだけでなく、高いブランク値と再現性のない結果を引き起こす危険性があります。11のため、オンチップのSPE技術のための戦略を働い代替は微量金属分析のために不可欠です。 1993年に、ワットChehimi 12は、金属イオンは、ポリマー材料に向けて保持する傾向を有することを見出し、そして分析物のほとんどは、効率的に、クロロ（CL）上に保持されていることを-containingのポリマー材料ナトリウムイオンを除いて、ポリ（塩化ビニル）（PVC）。そのため、2002年に、Eboatu ら 13は、さらにPVCによってソリューションからいくつかの有毒金属の隔離について報告しました。これは、検体の前濃縮および塩マトリックスの除去のための優れた特性を示した高分子材料をClを含有することを示したので、のCl含有SPE機能を備えたチップベースのデバイスは、決意のための新規のオンチップSPE技術の開発のための魅力的な戦略と考えられました微量金属イオン。そのような製造の容易さ、所望の化学的/機械的特性、および光学的透明性などを考慮すると材料の機能は、14,15この研究では、マイクロデバイスを製造するために、ポリ（メチルメタクリレート）（PMMA）を利用しました。そして、CL-含有SPE機能は、微量金属イオンの決意するための新規なオンチップSPE技術の開発のために作製した素子に注入した。16 Remarkably、チャネル内部の高度に電気陰性C-Clでの部分の間の双極子 - イオン相互作用および正に帯電した金属イオン上の革新的な抽出機構の依存はにつながる、一般的なオンチップのSPE手順の間に講じた措置を回避することができます過剰の試薬を使用することによって引き起こされる汚染や追加のステップに起因する労働者のいずれかの劇的な低下。この貢献で提供されたプロトコルは、自分の仕事のための双極子支援SPEのマイクロチップを製造するために多様な背景から研究者を可能にします。製造されたマイクロチップのための詳細な特徴付け手順も同様に記載されています。

<table><tbody><tr><td>AutoCAD</td><td>Autodesk</td><td>N/A</td><td>http://www.autodesk.com/education/free-software/autocad</td></tr><tr><td>Poly(methyl methacrylate) (PMMA) sheet</td><td>Kun Quan Engineering Plastics</td><td>N/A</td><td>350 mm (L) x 20 mm (W) x 2 mm (H). The glass transition temperature (Tg) of PMMA sheets is ranged from 102&amp;ndash;110 &amp;deg;C. The UV transmittance of the PMMA at 365 nm is 91.2%.</td></tr><tr><td>Micromachining system</td><td>Laser Life</td><td>LES-10</td><td>Maximum laser power: 10 W. Maximium engraving speed: 762 mm sec&lt;sup&gt;&amp;minus;1&lt;/sup&gt;.</td></tr><tr><td>High-resolution optical microscope</td><td>Ching Hsing Computer-Tech</td><td>FS-230</td><td></td></tr><tr><td>Power Image Analysis system (PIA)</td><td>Ching Hsing Computer-Tech</td><td>PIA V16.1</td><td></td></tr><tr><td>Multi drilling machines</td><td>N/A</td><td>LT-848</td><td></td></tr><tr><td>Deionized water (D. I. H&lt;sub&gt;2&lt;/sub&gt;O)</td><td>Millipore</td><td>Milli-Q Integral 5 System</td><td></td></tr><tr><td>Sodium dodecyl sulfate (SDS)</td><td>J. T. Baker</td><td>4095-04</td><td></td></tr><tr><td>Ultrasound oscillator</td><td>Elma</td><td>Transsonic Digital</td><td></td></tr><tr><td>Glass board</td><td>N/A</td><td>N/A</td><td>160 mm (L) x 35 mm (W) x 2 mm (H); fragile</td></tr><tr><td>Binder clip</td><td>SDI</td><td>0234T-1</td><td>http://stationery.sdi.com.tw/product_detail.php?Key=322&amp;amp;cID=55&amp;amp;uID=6</td></tr><tr><td>Precision oven</td><td>Yeong Shin</td><td>DK-45</td><td></td></tr><tr><td>Poly(etheretherketone) (PEEK) tube</td><td>VICI</td><td>JR-T-6002 (0.5 mm i.d.); JR-T-6001 (0.25 mm i.d.)</td><td></td></tr><tr><td>Polymer tubing&amp;nbsp; cutter</td><td>Upchurch Scientific</td><td>A-327</td><td></td></tr><tr><td>Two-component epoxy-based adhesive</td><td>Richwang</td><td>N/A</td><td>Skin irritative. The major components are an epoxy resin and a hardener.</td></tr><tr><td>Peristaltic pump</td><td>Gilson</td><td>Minipuls 3</td><td></td></tr><tr><td>Peristaltic tube</td><td>Gilson</td><td>F117934</td><td></td></tr><tr><td>Sodium hydroxide (NaOH)</td><td>Sigma&amp;ndash;Aldrich</td><td>30620</td><td></td></tr><tr><td>Nitric acid (HNO&lt;sub&gt;3&lt;/sub&gt;)</td><td>J. T. Baker</td><td>959834</td><td></td></tr><tr><td>Acrylamide (prop-2-enamide, C&lt;sub&gt;3&lt;/sub&gt;H&lt;sub&gt;5&lt;/sub&gt;NO)</td><td>Sigma&amp;ndash;Aldrich</td><td>A8887</td><td>Acutely toxic and carcinogenic</td></tr><tr><td>In-house-built photomask</td><td>N/A</td><td>N/A</td><td>The in-house-built photomask was made of a black paper (114 mm (L) &amp;times; 22 mm (W)) that contained an open window (94 mm (L) &amp;times; 2 mm (W)) allowing the desired region</td></tr><tr><td>1,1-Dichloroethylene</td><td>Sigma&amp;ndash;Aldrich</td><td>163032</td><td>Acutely toxic and carcinogenic</td></tr><tr><td>Cartridge</td><td>Dikma</td><td>ProElut AL-B&amp;nbsp;</td><td></td></tr><tr><td>2,2-Azobisisobutyronitrile (AIBN, C&lt;sub&gt;8&lt;/sub&gt;H&lt;sub&gt;12&lt;/sub&gt;N&lt;sub&gt;4&lt;/sub&gt;)</td><td>Showa Chemical&amp;nbsp;</td><td>0159-2130</td><td></td></tr><tr><td>Ethanol</td><td>Sigma&amp;ndash;Aldrich</td><td>32221</td><td></td></tr><tr><td>Hexanes (C&lt;sub&gt;6&lt;/sub&gt;H&lt;sub&gt;14&lt;/sub&gt;)</td><td>Millinckrodt Chemical</td><td>5189-08</td><td></td></tr><tr><td>In-house-built irradiation system</td><td>Great Lighting (UV-A lamp)</td><td>N/A</td><td>An opaque box with an UV-A lamp (40 W, maximum emission at 365 nm)</td></tr><tr><td>Glass vial</td><td>Yeong Shin</td><td>132300019</td><td>Fragile</td></tr><tr><td>Aluminum foil</td><td>Diamond</td><td>N/A</td><td></td></tr><tr><td>Conical tubes with screw caps</td><td>labcon</td><td>3181-345-008 (50 mL); 3131-345-008 (15 mL)</td><td></td></tr><tr><td>Rocking shaker</td><td>TKS</td><td>RS-01</td><td></td></tr><tr><td>Contact angle meter</td><td>First Ten Angstroms</td><td>FTA 125</td><td></td></tr><tr><td>PMMA bead</td><td>Scientific Polymer Products</td><td>037A</td><td></td></tr><tr><td>Mortar and pestle, agate&amp;nbsp;</td><td>Yeong Shin</td><td>139000004</td><td>Fragile</td></tr><tr><td>Tissue culture plate</td><td>AdvanGene Life Science Plasticware</td><td>AGC-CP-24S-50EA</td><td>24-Well, non-treated, sterilized</td></tr><tr><td>Hydraulic press</td><td>Panchum</td><td>Press-200</td><td></td></tr><tr><td>Laser ablation</td><td>New Wave Research</td><td>NWR193</td><td></td></tr><tr><td>Inductively coupled plasma-mass spectrometer</td><td>Agilent Technologies</td><td>Agilent 7500a</td><td></td></tr><tr><td>Glass bottle</td><td>DURAN</td><td>21801245 (100 mL); 21801365 (250 mL)</td><td></td></tr><tr><td>Dispersive Raman spectrometer</td><td>Thermo Fisher Scientific</td><td>Nicolet Almega XR</td><td></td></tr><tr><td>Manganese nitrate tetrahydrate (Mn(NO&lt;sub&gt;3&lt;/sub&gt;)&lt;sub&gt;2&lt;/sub&gt;&amp;times;4H&lt;sub&gt;2&lt;/sub&gt;O)</td><td>Sigma&amp;ndash;Aldrich</td><td>63547</td><td></td></tr><tr><td>Maleic acid disodium salt hydrate (C&lt;sub&gt;4&lt;/sub&gt;H&lt;sub&gt;4&lt;/sub&gt;Na&lt;sub&gt;2&lt;/sub&gt;O&lt;sub&gt;5&lt;/sub&gt;)</td><td>Sigma&amp;ndash;Aldrich</td><td>M9009</td><td></td></tr><tr><td>X-ray absorption near edge structure (XANES)&amp;nbsp;</td><td>N/A</td><td>N/A</td><td>The Mn K-edge XANES analyses were conducted at 07A and 17C1 beamlines of the National Synchrotron Radiation Research Center (NSRRC) in Taiwan.</td></tr></tbody></table>

fabrication of a dipole-assisted solid phase extraction microchip for trace metal analysis in water samples

本論文では、水試料中の微量金属分析のために利用可能な双極子支援固相抽出（SPE）マイクロチップの製造プロトコルについて説明します。チップベースのSPE技術の進化の概要が提供されます。これは、特定のポリマー材料への導入とSPEにおけるその役割が続いています。革新的な双極子補助SPE法を開発するために、SPE機能を含有塩素（CL）は、ポリ（メチルメタクリレート）（PMMA）マイクロチップに注入しました。ここで、接触角分析、ラマン分光分析、およびレーザーアブレーション、誘導結合プラズマ質量分析法を含む様々な分析技術（LA-ICP-MS）分析は、上のC-Clで部分の注入プロトコルの有用性を検証するために使用しましたPMMA。 X線吸収端近傍構造解析結果（XANES）分析もまた、双極子によって抽出媒体としてのCl含有PMMAの実現可能性を実証しました高度に電気陰性C-Clで部分と正に帯電した金属イオンとイオンの相互作用。

図2は、PMMAマイクロチップのチャネル変更の手続きの際に発生する反応を示しています。接触角分析は、提案されている手順の間、表面の変更を監視するために使用しました。 LA-ICP-MSシステムおよび分散型ラマン分光計はC-Clでの成功した変更を確認するために使用されたがPMMA基板上に形成する部分（ 図3の（a）、（b）参照 ）。提案された双極子補助SPE反応はXANES解析（ 図4）によって特徴付けました。 <img alt="図1" src="/files/ftp_upload/53500/53500fig1.jpg" /> 図1. PMMAマイクロチップ。（a） の製作マイクロチップ用のパターンファイルのスナップショット。 （b）は製作マイクロチップのレイアウト：S、E、およびBは、試料のための導入ポートを表し、溶離液、およびBUFをそれぞれの溶液をFER。 Oは、出口を表しています。黒丸はそれぞれのための掘削のアクセスホールを表します。サンプルおよびバッファソリューションの導入に使用されるチャネルの両方が抽出チャネルと30°の角度を形成しました。コンフルエント出口に試料と緩衝液の流れの収束点からの距離とした効果的な抽出流路の長さは94 mmでした。 （c）は 、機械加工板の断面写真。文献から再生。 16化学の王立協会の許可を得て。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53500/53500fig1large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> <img alt="図2" src="/files/ftp_upload/53500/53500fig2.jpg" /> PMMAマイクロチップのためのチャネル変更の 図2. スキーム。差し込みphotogrAPHSシーケンスで得られた生成物に対応する接触角を示します。接触角は、水滴の画像を用いて決定しました。 3回の繰り返し測定の平均は、それぞれの場合において報告接触角を測定するために使用しました。文献から再生。 16化学の王立協会の許可を得て。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53500/53500fig2large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> <img alt="図3" src="/files/ftp_upload/53500/53500fig3.jpg" /> PMMAの変更の 図3. 表面検証。（A）のClのための信号がC-Clでの部分で修飾されたPMMA及びPMMAの両方を切除することにより得られます。挿入図は、各取得信号に対応する切除位置を示しています。 （b）は 、ネイティブのラマンスペクトルとPMMAを変更しました。文献から再生。の許可を得て16化学の王立協会。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53500/53500fig3large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> <img alt="図4" src="/files/ftp_upload/53500/53500fig4.jpg" /> 修正されたPMMA及びMn 2+ イオン で ​​処理された修正されたPMMAの 図4 のMnのK端XANESスペクトル 。修正されたPMMAのスペクトルは赤線のように発表されました。吸収スペクトルを示す変形PMMAの高度に電気陰性C-Clで部分とMn 2+イオンとの間の相互作用は、青色の線として提示されました。文献から再生。 16化学の王立協会の許可を得て。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/53500/53500fig4large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a>

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Fabrication of a Dipole-assisted Solid Phase Extraction Microchip for Trace Metal Analysis in Water Samples

The fabrication protocol of a dipole-assisted solid phase extraction microchip for the trace metal analysis is presented.

水試料中の微量金属分析のためのダイポールアシスト固相抽出マイクロチップの作製

This paper describes a fabrication protocol for a dipole-assisted solid phase extraction (SPE) microchip available for trace metal analysis in water ...

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8.6K ビュー.  National Tsing Hua University. The fabrication protocol of a dipole-assisted solid phase extraction microchip for the trace metal analysis is presented.

ビデオ: Fabrication of a Dipole-assisted Solid Phase Extraction Microchip for Trace Metal Analysis in Water Samples

A Simple Method for Automated Solid Phase Extraction of Water Samples for Immunological Analysis of Small Pollutants

A new method for solid phase extraction (SPE) of environmental water samples is proposed. The developed prototype is cost-efficient and user friendly, and enables to perform rapid, automated and simple SPE. The pre-concentrated solution is compatible with analysis by immunoassay, with a low organic solvent content. A method is described for the extraction and pre-concentration of natural hormone 17&#946;-estradiol in 100 ml water samples. Reverse phase SPE is performed with octadecyl-silica sorbent and elution is done with 200 &#181;l of methanol 50% v/v. Eluent is diluted by adding di-water to lower the amount of methanol. After preparing manually the SPE column, the overall procedure is performed automatically within 1 hr. At the end of the process, estradiol concentration is measured by using a commercial enzyme-linked immune-sorbent assay (ELISA). 100-fold pre-concentration is achieved and the methanol content in only 10% v/v. Full recoveries of the molecule are achieved with 1 ng/L spiked de-ionized and synthetic sea water samples.

A protocol for the extraction and pre-concentration of estradiol from water samples by using an automated and miniaturized system is presented.

小さな汚染物質の免疫学的分析のための水試料の自動固相抽出ための簡単​​な方法

A new method for solid phase extraction (SPE) of environmental water samples is proposed. The developed prototype is cost-efficient and user friendly, and ...

環境学

Clean Sampling and Analysis of River and Estuarine Waters for Trace Metal Studies

Most of the trace metal concentrations in ambient waters obtained a few decades ago have been considered unreliable owing to the lack of contamination control. Developments of some techniques aiming to reduce trace metal contamination in the last couple of decades have resulted in concentrations reported now being orders of magnitude lower than those in the past. These low concentrations often necessitate preconcentration of water samples prior to instrumental analysis of samples. Since contamination can appear in all phases of trace metal analyses, including sample collection (and during preparation of sampling containers), storage and handling, pretreatments, and instrumental analysis, specific care needs to be taken in order to reduce contamination levels at all steps. The effort to develop and utilize "clean techniques" in trace metal studies allows scientists to investigate trace metal distributions and chemical and biological behavior in greater details. This advancement also provides the required accuracy and precision of trace metal data allowing for environmental conditions to be related to trace metal concentrations in aquatic environments.
This protocol that is presented here details needed materials for sample preparation, sample collection, sample pretreatment including preconcentration, and instrumental analysis. By reducing contamination throughout all phases mentioned above for trace metal analysis, much lower detection limits and thus accuracy can be achieved. The effectiveness of "clean techniques" is further demonstrated using low field blanks and good recoveries for standard reference material. The data quality that can be obtained thus enables the assessment of trace metal distributions and their relationships to environmental parameters.

Special care using "clean techniques" is required to properly collect and process water samples for trace metal studies in aquatic environments. A protocol for sampling, processing, and analytical procedures with the aim of obtaining reliable environmental monitoring data and results with high sensitivity for detailed trace metal studies is presented.

きれいなサンプリングと微量金属の研究のために川や河口ウォーターズの分析

Most of the trace metal concentrations in ambient waters obtained a few decades ago have been considered unreliable owing to the lack of contamination ...

On-chip Isotachophoresis for Separation of Ions and Purification of Nucleic Acids

Electrokinetic techniques are a staple of microscale applications because of their unique ability to perform a variety of fluidic and electrophoretic processes in simple, compact systems with no moving parts. Isotachophoresis (ITP) is a simple and very robust electrokinetic technique that can achieve million-fold preconcentration1,2 and efficient separation and extraction based on ionic mobility.3 For example, we have demonstrated the application of ITP to separation and sensitive detection of unlabeled ionic molecules (e.g. toxins, DNA, rRNA, miRNA) with little or no sample preparation4-8 and to extraction and purification of nucleic acids from complex matrices including cell culture, urine, and blood.9-12
ITP achieves focusing and separation using an applied electric field and two buffers within a fluidic channel system. For anionic analytes, the leading electrolyte (LE) buffer is chosen such that its anions have higher effective electrophoretic mobility than the anions of the trailing electrolyte (TE) buffer (Effective mobility describes the observable drift velocity of an ion and takes into account the ionization state of the ion, as described in detail by Persat et al.13). After establishing an interface between the TE and LE, an electric field is applied such that LE ions move away from the region occupied by TE ions. Sample ions of intermediate effective mobility race ahead of TE ions but cannot overtake LE ions, and so they focus at the LE-TE interface (hereafter called the "ITP interface"). Further, the TE and LE form regions of respectively low and high conductivity, which establish a steep electric field gradient at the ITP interface. This field gradient preconcentrates sample species as they focus. Proper choice of TE and LE results in focusing and purification of target species from other non-focused species and, eventually, separation and segregation of sample species.
We here review the physical principles underlying ITP and discuss two standard modes of operation: "peak" and "plateau" modes. In peak mode, relatively dilute sample ions focus together within overlapping narrow peaks at the ITP interface. In plateau mode, more abundant sample ions reach a steady-state concentration and segregate into adjoining plateau-like zones ordered by their effective mobility. Peak and plateau modes arise out of the same underlying physics, but represent distinct regimes differentiated by the initial analyte concentration and/or the amount of time allotted for sample accumulation.
We first describe in detail a model peak mode experiment and then demonstrate a peak mode assay for the extraction of nucleic acids from E. coli cell culture. We conclude by presenting a plateau mode assay, where we use a non-focusing tracer (NFT) species to visualize the separation and perform quantitation of amino acids.

Isotachophoresis (ITP) is a robust electrokinetic separation and preconcentration technique with applications ranging from toxin detection to sample preparation. We review the physical principles of ITP and the methodology of applying this technique to two specific example applications: separation and detection of small molecules and purification of nucleic acids from cell culture lysate.

イオンの分離及び核酸の精製のためのオンチップ等速電気泳動

Electrokinetic techniques are a staple of microscale applications because of their unique ability to perform a variety of fluidic and electrophoretic ...

生物工学

Detection and Recovery of Palladium, Gold and Cobalt Metals from the Urban Mine Using Novel Sensors/Adsorbents Designated with Nanoscale Wagon-wheel-shaped Pores

Developing low-cost, efficient processes for recovering and recycling palladium, gold and cobalt metals from urban mine remains a significant challenge in industrialized countries. Here, the development of optical mesosensors/adsorbents (MSAs) for efficient recognition and selective recovery of Pd(II), Au(III), and Co(II) from urban mine was achieved. A simple, general method for preparing MSAs based on using high-order mesoporous monolithic scaffolds was described. Hierarchical cubic Ia3d wagon-wheel-shaped MSAs were fabricated by anchoring chelating agents (colorants) into three-dimensional pores and micrometric particle surfaces of the mesoporous monolithic scaffolds. Findings show, for the first time, evidence of controlled optical recognition of Pd(II), Au(III), and Co(II) ions and a highly selective system for recovery of Pd(II) ions (up to ~95%) in ores and industrial wastes. Furthermore, the controlled assessment processes described herein involve evaluation of intrinsic properties (e.g., visual signal change, long-term stability, adsorption efficiency, extraordinary sensitivity, selectivity, and reusability); thus, expensive, sophisticated instruments are not required. Results show evidence that MSAs will attract worldwide attention as a promising technological means of recovering and recycling palladium, gold and cobalt metals.

Because of the importance and extensive use of palladium, gold and cobalt metals in high-technology equipment, their recovery and recycling constitute an important industrial challenge. The metal recovery system described herein is a simple, low-cost means for the effective detection, removal, and recovery of these metals from the urban mine.

検出とパラジウムの回収は、都市鉱山からの金やコバルト金属は、ナノスケールワゴンホイール形の細孔を有する指定小説センサ/吸着剤を使用しました

Developing low-cost, efficient processes for recovering and recycling palladium, gold and cobalt metals from urban mine remains a significant challenge in ...

工学

Fabrication of the Thermoplastic Microfluidic Channels

In our lab, we have successfully isolated nucleic acids directly from microliter and submicroliter volumes of human blood, urine and stool using polymer/nanoparticle composite microscale lysis and solid phase extraction columns. The recovered samples are concentrated, small volume samples that are PCRable, without any additional cleanup. Here, we demonstrate how to fabricate thermoplastic microfluidic chips using hot embossing and heat sealing. Then, we demonstrate how to use in situ light directed surface grafting and polymerization through the sealed chip to form the composite solid phase columns. We demonstrate grafting and polymerization of a carbon nanotube/polymer composite column for bacterial cell lysis. We then show the lysis process followed by solid phase extraction of nucleic acids from the sample on chip using a silica/polymer composite column. The attached protocols contain detailed instructions on how to make both lysis and solid phase extraction columns.

Here we demonstrate how to fabricate thermoplastic microfluidic chips using hot embossing and heat sealing. Then we demonstrate how to use in situ light directed surface grafting and polymerization through the sealed chip to form the composite solid phase columns.

熱可塑性マイクロ流体チャネルの作製

In our lab, we have successfully isolated nucleic acids directly from microliter and submicroliter volumes of human blood, urine and stool using ...

生物学

A Microfluidic Chip for the Versatile Chemical Analysis of Single Cells

We present a microfluidic device that enables the quantitative determination of intracellular biomolecules in multiple single cells in parallel. For this purpose, the cells are passively trapped in the middle of a microchamber. Upon activation of the control layer, the cell is isolated from the surrounding volume in a small chamber. The surrounding volume can then be exchanged without affecting the isolated cell. However, upon short opening and closing of the chamber, the solution in the chamber can be replaced within a few hundred milliseconds. Due to the reversibility of the chambers, the cells can be exposed to different solutions sequentially in a highly controllable fashion, e.g. for incubation, washing, and finally, cell lysis. The tightly sealed microchambers enable the retention of the lysate, minimize and control the dilution after cell lysis. Since lysis and analysis occur at the same location, high sensitivity is retained because no further dilution or loss of the analytes occurs during transport. The microchamber design therefore enables the reliable and reproducible analysis of very small copy numbers of intracellular molecules (attomoles, zeptomoles) released from individual cells. Furthermore, many microchambers can be arranged in an array format, allowing the analysis of many cells at once, given that suitable optical instruments are used for monitoring. We have already used the platform for proof-of-concept studies to analyze intracellular proteins, enzymes, cofactors and second messengers in either relative or absolute quantifiable manner.

In this article we present a microfluidic chip for single cell analysis. It allows the quantification of intracellular proteins, enzymes, cofactors, and second messengers by means of fluorescent assays or immunoassays.&#160;

単一細胞の多彩な化学分析マイクロ流体チップ

We present a microfluidic device that enables the quantitative determination of intracellular biomolecules in multiple single cells in parallel. For this ...

A Microfluidic-based Electrochemical Biochip for Label-free DNA Hybridization Analysis

Miniaturization of analytical benchtop procedures into the micro-scale provides significant advantages in regards to reaction time, cost, and integration of pre-processing steps. Utilizing these devices towards the analysis of DNA hybridization events is important because it offers a technology for real time assessment of biomarkers at the point-of-care for various diseases. However, when the device footprint decreases the dominance of various physical phenomena increases. These phenomena influence the fabrication precision and operation reliability of the device. Therefore, there is a great need to accurately fabricate and operate these devices in a reproducible manner in order to improve the overall performance. Here, we describe the protocols and the methods used for the fabrication and the operation of a microfluidic-based electrochemical biochip for accurate analysis of DNA hybridization events. The biochip is composed of two parts: a microfluidic chip with three parallel micro-channels made of polydimethylsiloxane (PDMS), and a 3 x 3 arrayed electrochemical micro-chip. The DNA hybridization events are detected using electrochemical impedance spectroscopy (EIS) analysis. The EIS analysis enables monitoring variations of the properties of the electrochemical system that are dominant at these length scales. With the ability to monitor changes of both charge transfer and diffusional resistance with the biosensor, we demonstrate the selectivity to complementary ssDNA targets, a calculated detection limit of 3.8 nM, and a 13% cross-reactivity with other non-complementary ssDNA following 20 min&#160;of incubation. This methodology can improve the performance of miniaturized devices by elucidating on the behavior of diffusion at the micro-scale regime and by enabling the study of DNA hybridization events.

We present a microfluidic-based electrochemical biochip for DNA hybridization detection. Following ssDNA probe functionalization, the specificity, sensitivity, and detection limit are studied with complementary and non-complementary ssDNA targets. Results illustrate the influence of the DNA hybridization events on the electrochemical system, with a detection limit of 3.8 nM.

ラベルフリーDNAハイブリダイゼーション分析のためのマイクロ流体ベースの電気化学バイオチップ

Miniaturization of analytical benchtop procedures into the micro-scale provides significant advantages in regards to reaction time, cost, and integration ...

A Microfluidic Chip for ICPMS Sample Introduction

This protocol discusses the fabrication and usage of a disposable low cost microfluidic chip as sample introduction system for inductively coupled plasma mass spectrometry (ICPMS). The chip produces monodisperse aqueous sample droplets in perfluorohexane (PFH). Size and frequency of the aqueous droplets can be varied in the range of 40 to 60 &#181;m and from 90 to 7,000&#160;Hz, respectively. The droplets are ejected from the chip with a second flow of PFH and remain intact during the ejection. A custom-built desolvation system removes the PFH and transports the droplets into the ICPMS. Here, very stable signals with a narrow intensity distribution can be measured, showing the monodispersity of the droplets. We show that the introduction system can be used to quantitatively determine iron in single bovine red blood cells. In the future, the capabilities of the introduction device can easily be extended by the integration of additional microfluidic modules.

We present a discrete droplet sample introduction system for inductively coupled plasma mass spectrometry (ICPMS). It is based on a cheap and disposable microfluidic chip that generates highly monodisperse droplets in a size range of 40&#8722;60 &#181;m at frequencies from 90 to 7,000 Hz.

ICPMSサンプル導入のためのマイクロ流体チップ

This protocol discusses the fabrication and usage of a disposable low cost microfluidic chip as sample introduction system for inductively coupled plasma ...

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. ...

Fast Enzymatic Processing of Proteins for MS Detection with a Flow-through Microreactor

The vast majority of mass spectrometry (MS)-based protein analysis methods involve an enzymatic digestion step prior to detection, typically with trypsin. This step is necessary for the generation of small molecular weight peptides, generally with MW &#60; 3,000-4,000 Da, that fall within the effective scan range of mass spectrometry instrumentation. Conventional protocols involve O/N enzymatic digestion at 37 &#186;C. Recent advances have led to the development of a variety of strategies, typically involving the use of a microreactor with immobilized enzymes or of a range of complementary physical processes that reduce the time necessary for proteolytic digestion to a few minutes (e.g., microwave or high-pressure). In this work, we describe a simple and cost-effective approach that can be implemented in any laboratory for achieving fast enzymatic digestion of a protein. The protein (or protein mixture) is adsorbed on C18-bonded reversed-phase high performance liquid chromatography (HPLC) silica particles preloaded in a capillary column, and trypsin in aqueous buffer is infused over the particles for a short period of time. To enable on-line MS detection, the tryptic peptides are eluted with a solvent system with increased organic content directly in the MS ion source. This approach avoids the use of high-priced immobilized enzyme particles and does not necessitate any aid for completing the process. Protein digestion and complete sample analysis can be accomplished in less than ~3 min and ~30 min, respectively.

A quick protocol for proteolytic digestion with an in-house built flow-through tryptic microreactor coupled to an electrospray ionization (ESI) mass spectrometer is presented. The fabrication of the microreactor, the experimental setup and the data acquisition process are described.

フロースルーマイクロリアクターとMS検出のためのタンパク質の高速酵素処理

The vast majority of mass spectrometry (MS)-based protein analysis methods involve an enzymatic digestion step prior to detection, typically with trypsin. ...

水試料中の微量金属分析のためのダイポールアシスト固相抽出マイクロチップの作製

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この動画の章

小さな汚染物質の免疫学的分析のための水試料の自動固相抽出ための簡単な方法

きれいなサンプリングと微量金属の研究のために川や河口ウォーターズの分析

イオンの分離及び核酸の精製のためのオンチップ等速電気泳動

検出とパラジウムの回収は、都市鉱山からの金やコバルト金属は、ナノスケールワゴンホイール形の細孔を有する指定小説センサ/吸着剤を使用しました

熱可塑性マイクロ流体チャネルの作製

単一細胞の多彩な化学分析マイクロ流体チップ

ラベルフリーDNAハイブリダイゼーション分析のためのマイクロ流体ベースの電気化学バイオチップ

ICPMSサンプル導入のためのマイクロ流体チップ

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

フロースルーマイクロリアクターとMS検出のためのタンパク質の高速酵素処理

水試料中の微量金属分析のためのダイポールアシスト固相抽出マイクロチップの作製

要約

さらに動画を探す

この動画の章

小さな汚染物質の免疫学的分析のための水試料の自動固相抽出ための簡単​​な方法

きれいなサンプリングと微量金属の研究のために川や河口ウォーターズの分析

イオンの分離及び核酸の精製のためのオンチップ等速電気泳動

検出とパラジウムの回収は、都市鉱山からの金やコバルト金属は、ナノスケールワゴンホイール形の細孔を有する指定小説センサ/吸着剤を使用しました

熱可塑性マイクロ流体チャネルの作製

単一細胞の多彩な化学分析マイクロ流体チップ

ラベルフリーDNAハイブリダイゼーション分析のためのマイクロ流体ベースの電気化学バイオチップ

ICPMSサンプル導入のためのマイクロ流体チップ

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

フロースルーマイクロリアクターとMS検出のためのタンパク質の高速酵素処理

小さな汚染物質の免疫学的分析のための水試料の自動固相抽出ための簡単な方法