我々は、NSFの賞CMMI - 1000724の資金援助を認める。

1A。シリコン上の主要な単分子膜形成<ol><li> 1センチメートル2基板、粉々にシリコンウエハーをカットし、水とフィルタリングエタノールですすいでください。 </li><li> 75℃でナノストリップを含むガラス皿の中でシリコン基板を沈めることによって有機汚染を取り除く15分後、脱イオン、ろ過水を使用して各基板をすすぐ。 </li><li>自然酸化層を除去するために：5％HF溶液（HFは非常に危険な物質である警告）に各基板を置きます。窒素で5分後にドライ酸化、フリーシリコン</li><li>すぐに水没クロロベンゼンの飽和PCL 5の2 mlを入れたシンチレーションバイアル内の各酸化物を含まないシリコン片を、塩素化基板を生産する。この溶液を0.2μmにフィルタリングする必要があります。 </li><li>各バイアルの上にバイアルのコンデンサーをアセンブルし、heatblockに配置は、112℃まで一時間を設定します。 </li><li>反応が完了した後、バイアルは、各surfaを冷却し、リンスしましょうろ過した窒素下でクロロベンゼンと乾燥とCE。 </li><li>プロペニル - 終了基板を形成するために、プロペニル塩化マグネシウムの4 mlを含む圧力のバイアル内の各塩素化シリコン表面に置きます。 130℃heatblock内の各圧力のバイアルを置き℃で24時間。 </li><li> heatblockの各圧力のバイアルを取り出し、冷ます。 </li><li> DCMとエタノールで迅速に各表面を洗浄し、ろ過した窒素下で乾燥させます。 </li></ol> 1B。ゲルマニウムのプライマリ単分子膜形成<ol><li> 、1平方センチメートル基板にゲルマニウムのウェーハをカット塵や水とフィルタリングエタノールですすいでください。 </li><li> 20分間アセトンを含有するガラス皿の中で表面を沈めることによって有機汚染を取り除く</li><li> 15分間10％HCl溶液で各表面に置きます。このプロセスは、同時に自然酸化層を除去し、表面をchlorinates。 5分後、窒素で基板を乾燥させます。 </li><li>オクチル-終端基板、ナンプラーを形成するCEは、各オクチル塩化マグネシウム（2mM）を4 mlを含む圧力のバイアルにゲルマニウム表面を塩素化。 ℃で48時間130℃heatblock内の各圧力のバイアルを置きます。 </li><li> heatblockの各圧力のバイアルを取り出し、室温まで冷ます。 </li><li> DCMとエタノールで迅速に各表面を洗浄し、ろ過した窒素下で乾燥させます。 </li></ol> 2。シリコンとゲルマニウムでNHSの基板の機能化<ol><li>四塩化炭素でフィルタリングさ0.1MのNHS -ジアジリンの溶液を調製します。警告：最小の露光をキープ。 </li><li>メチル終端表面上にピペットで溶液を数滴。表面全体に分散するためのソリューションを許可する。 </li><li> UVランプ（☐= 254 nmの、0.74インチで4400/cm2）で表面を置きます。表面が表面に多くのNHS -ジアジリンを追加し、30分間UV光下で反応し、さらに30分間反応反応を聞かせすることができます。 </li><li> NHS変更されたのをすすぐDCMとエタノールと、ろ過した窒素下で乾燥とurfaces。 </li></ol> 3。小分子の機能化<ol><li>反応するNHS修飾20mMのtert -ブチルカルバモイル、室温で2時間ジクロロメタン（DCM）におけるカルボニル（Boc -）エチレンジアミンの溶液中に基板を。 </li><li>反応後、DCMとエタノールとのBoc -修飾基質をすすいでください。 </li><li>室温で1時間、DCMには25％トリフルオロ酢酸（TFA）を用いてBoc変更された基板を脱保護。 </li><li>ろ過した窒素下で水と乾燥で、その結果DCMと表面、エタノール、10％（w / v）の炭酸水素カリウムをすすぐ。 </li><li>元素組成を決定するためにXPSにより、すべての面を分析する。 </li></ol> 4。酸性ポリウレタンアクリレートスタンプ（PUA）の準備<ol><li>粘度を減らすためにトリメチロールプロパンエトキシレートトリアクリレートBで30％、アクリル酸を希釈する。反応混合物（ 図に光開始剤CとDを追加します。URE 6）。 </li><li>ジオキサン中の4N HCl溶液（10ml）に、2分間室温で撹拌ナトリウム2 - mercaptoethanesulfonate（0.2グラム、1.22ミリモル）を追加します。 </li><li>細かいガラスフィルターで、次にジオキサン、2 - mercaptoethanesulfonic酸の透明な溶液を得たフィルタ0.2μメートルのPTFEメンブレンシリンジを介して第塩化ナトリウムをフィルタオフ。 </li><li>減圧下でジオキサンを蒸発させる</li><li> 50℃室温でポリウレタンアクリレートprepolymeric混合物2mlで、その後真空下で、得られたスルホン酸を反応℃に完全に閉じ込められた空気の泡から混合物を解放するようにしてください。 </li><li>室温に得られた溶液を冷却し、二つのガラス顕微鏡スライドまたは室温で2時間のUV光への曝露によるガラスのスライドとマスターの間で重合する。 </li><li>重合後に、慎重にマスターオフタイムスタンプピールとフィルタリングnitrogeでエタノールと水でスタンプを洗浄し、乾燥nの</li></ol> 5。触媒印刷とSEM / AFM解析<ol><li>それらを一緒に保持するために、外部負荷で1分間室温でNHS -修飾基質の上に対応するポリウレタンアクリレートスタンプを置きます。 </li><li>反応後、スタンプと基板を分離する。 </li><li>その後、エタノール、水、エタノールで、ろ過した窒素で乾燥した基板を洗浄します。 </li><li>その後、エタノール、水、エタノールで、ろ過した窒素とドライスタンプをすすいでください。 </li><li>次のアプリケーションの前に室温でスタンプしてください。 </li><li>接触モードの横方向の原子間力microsopy（AFM）および走査型電子顕微鏡（SEM）を使用して生成したパターンを分析する</li></ol> 6。タンパク質のパターニングと蛍光顕微鏡<ol><li>ですすいだ後、1時間室温でH20（1:1）と：水没リジン- N、N -二酢酸（20 mm）とDMF中のEt 3 N（100 mM）の中のNHS柄二官能性基質を水とエタノール。 </li><li>室温で5分間、50mMのNiSO4溶液中の基質をインキュベートする。 </li><li> 0で1時間フィルタリングGFPの溶液中で水と結合バッファー（20mMのNAP、250mMのNaCl、10mMのイミダゾール、pH7.5）および水没（〜40μM）で過度にキレート化基材を洗い流す℃に</li><li>直ちにPBS（pH7.4）に続いて結合バッファーと基質をすすいでください。 </li><li>彼らは蛍光顕微鏡分析のための準備ができるまで、0℃でPBS中に基板が水和してください。 </li></ol> 7。タンパク質のパターニングと蛍光顕微鏡<ol><li>水ですすいだ後、H 2 1時間室温で0（1:1）と：リジン- N、N -二酢酸（20 mm）とDMF中のEt 3 N（100 mm）の水没NHS柄の二官能性基質とエタノール。 </li><li>室温で5分間、50mMのNISO 4水溶液に基板をインキュベートする。 </li><li>過度にワットキレート基板をすすぐ0で1時間のフィルタGFPの溶液中でi番目の水と結合バッファー（20mMのNAP、250mMのNaCl、10mMのイミダゾール、pH7.5）および水没（〜40μM）℃ </li><li>直ちにPBS（pH7.4）に続いて結合バッファーと基質をすすいでください。 </li><li>彼らは蛍光顕微鏡分析のための準備ができるまで、0℃でPBS中に基板が水和してください。 </li></ol> 8。代表的な結果： ソフトリソグラフィ触媒ナノパターニングの例を図7に示します。アプローチは、直角に異なる化学的および生物学的な部分で官能化することができる酸化物を含まないシリコンとゲルマニウム、上の化学選択的なパターンを作成します。 NHS - functioanlized基板と触媒パターニングスタンプとの間の反応では、NHS活性化し、遊離カルボン酸のパターン化二官能性基質のベアリングの領域をもたらし、コンフォーマル接触の分野におけるNHSの部分の加水分解につながります。 diffusによる本手法のイオンフリーの性質は、我々はフォトリソグラフィに近い分解能を達成。例えば、 図7は、全体に均一シリコン基板の表面全体に再現された125 nmの特徴を示しています。驚くべきことに、触媒スタンプが効率を失うことなく、複数回再利用することができます。 生体分子とのパターン化された半導体の化学選択的官能基は、センシング、診断、および研究の分析の分野におけるアプリケーション向けに高度に選択的な生物学的基質との伝統的な電子材料を統合の展望を開きます。このような官能の例は、NHS -パターニングシリコンが選択的に蛋白質分子で機能化された図8に示されています。活性化し、遊離カルボン酸の差反応性を悪用することによって、我々は最初のNHS官能地域にニトリロ三酢酸で終了する（NTA）ヘテロリンカーを貼付して、結果の使用ヘキサヒスチジンタグGFP 図8bの選択的付着のためのテンプレートとしてNTA -パターン表面は明らかにGFP - Modifiedと加水分解された遊離カルボン酸の地域間の差蛍光強度を示しています。複製された機能の大きさと形状は炭素動態化表面とスタンピングアプローチの選択性の顕著な安定性を確認し、NHSパターン表面（ 図8a）とGFP変性表面（ 図8b）の両方の間で一貫しています。プロトコルは、Hisタグ融合タンパク質に限定されるものではなく、DNAと抗体を含むパターン、他の生体分子に使用することができます。 <img src="/files/ftp_upload/3478/3478fig1.jpg" alt="図1"/> 図1。触媒マイクロコンタクトプリンティングを表す一般的なスキーム<img src="/files/ftp_upload/3478/3478fig2.jpg" alt="図2"/> 図2二層mの構造GeとSi基板上olecularシステム。主なアルキル単分子膜は、基板との安定したGeがCもしくはSi - C結合を形成し、基底サーフェス劣化からの保護、化学的に不活性と密接なパックシステムを提供する。（b）の二次被覆層が主な保護層で安定したCC結合を形成し、機能的な端末を提供するグループ<img src="/files/ftp_upload/3478/3478fig3.jpg" alt="図3"/> Siの（A）とGe（B）の一次防護単分子膜の形成を表す図3。反応スキーム<img src="/files/ftp_upload/3478/3478fig4.jpg" alt="図4"/> 図4。ヘテロカルベンのドナーとの一次防護単層の化学的機能化<img src="/files/ftp_upload/3478/3478fig5.jpg" alt="図5"/> 図5。NHS官能サブの低分子化合物の変更を示す反応スキームstratesと対応するXPSスペクトル<img src="/files/ftp_upload/3478/3478fig6.jpg" alt="図6"/> 図6。触媒プレポリマー混合物、重合条件、およびパターンスルホン酸変性スタンプのSEM像の構成と対応するPMMA - Siのマスター<img src="/files/ftp_upload/3478/3478fig7.jpg" alt="図7"/> 図7。酸性スタンプ付きSEMとSiとGe上にパターン化SAMのAFMの摩擦の画像<img src="/files/ftp_upload/3478/3478fig8.jpg" alt="図8"/> 図8有機及び生体分子との不動態化されたシリコンのソフトリソグラフィパターニングと機能化：。。パターニングされたNHS -修飾基質のSEM像B：GFP修飾基質の蛍光顕微鏡写真。

<ol>
	<li>Kumar, A., Abbott, N. L., Kim, E., Biebuyck, H. A., Whitesides, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patterned+Self-Assembled+Monolayers+and+Mesoscale+Phenomena.">Patterned Self-Assembled Monolayers and Mesoscale Phenomena.</a> Accounts. Chem. Res. 28 (5), 219-226 (1995).</li><li>Kumar, A., Biebuyck, H. A., Whitesides, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patterning+Self-Assembled+Monolayers:+Applications+in+Materials+Science.">Patterning Self-Assembled Monolayers: Applications in Materials Science.</a> Langmuir. 10 (5), 1498-1511 (1994).</li><li>Kumar, A., Whitesides, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Features+of+gold+having+micrometer+to+centimeter+dimensions+can+be+formed+through+a+combination+of+stamping+with+an+elastomeric+stamp+and+an+alkanethiol+"ink"+followed+by+chemical+etching.">Features of gold having micrometer to centimeter dimensions can be formed through a combination of stamping with an elastomeric stamp and an alkanethiol "ink" followed by chemical etching.</a> Applied Physics Letters. 63 (14), 2002-2004 (1993).</li><li>Wilbur, J. L., Kumar, A., Biebuyck, H. A., Kim, E., Whitesides, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microcontact+printing+of+self-assembled+monolayers:+applications+in+microfabrication.">Microcontact printing of self-assembled monolayers: applications in microfabrication.</a> Nanotechnology. 7 (4), 452-457 (1996).</li><li>Wilbur, J. L., Kumar, A., Kim, E., Whitesides, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfabrication+by+microcontact+printing+of+self-assembled+monolayers.">Microfabrication by microcontact printing of self-assembled monolayers.</a> Advanced Materials (Weinheim, Germany). 6 (7/8), 600-604 (1994).</li><li>Ruiz, S. A., Chen, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microcontact+printing:+a+tool+to+pattern.">Microcontact printing: a tool to pattern.</a> Soft Matter. 3 (2), 168-177 (2007).</li><li>Perl, A., Reinhoudt, D. N., Huskens, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microcontact+Printing:+Limitations+and+Achievements.">Microcontact Printing: Limitations and Achievements.</a> Advanced Materials (Weinheim, Germany). 21 (22), 2257-2268 (2009).</li><li>Kumar, A., Biebuyck, H. A., Abbott, N. L., Whitesides, G. 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+use+of+self-assembled+monolayers+and+a+selective+etch+to+generate+patterned+gold+features.">The use of self-assembled monolayers and a selective etch to generate patterned gold features.</a> J. Am. Chem. Soc. 114 (23), 9188-9191 (1992).</li><li>Ravoo, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microcontact+chemistry:+surface+reactions+in+nanoscale+confinement.">Microcontact chemistry: surface reactions in nanoscale confinement.</a> Journal of Materials Chemistry. 19 (47), 8902-8906 (2009).</li><li>Biebuyck, H. A., Larsen, N. B., Delamarche, E., Michel, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lithography+beyond+light:+Microcontact+printing+with+monolayer+resists.">Lithography beyond light: Microcontact printing with monolayer resists.</a> Ibm. J. Res. Dev. 41 (1-2), 159-170 (1997).</li><li>Delamarche, E., Schmid, H., Bietsch, A., Larsen, N. B., Rothuizen, H., Michel, B., Biebuyck, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transport+Mechanisms+of+Alkanethiols+during+Microcontact+Printing+on+Gold.">Transport Mechanisms of Alkanethiols during Microcontact Printing on Gold.</a> J. Phys. Chem. B. 102 (18), 3324-3334 (1998).</li><li>Larsen, N. B., Biebuyck, H., Delamarche, E., Michel, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Order+in+microcontact+printed+self-assembled+monolayers.">Order in microcontact printed self-assembled monolayers.</a> J Am Chem Soc. 119 (13), 3017-3026 (1997).</li><li>Michel, B., Bernard, A., Bietsch, A., Delamarche, E., Geissler, M., Juncker, D., Kind, H., Renault, J. P., Rothuizen, H., Schmid, H., Schmidt-Winkel, P., Stutz, R., Wolf, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Printing+meets+lithography:+Soft+approaches+to+high-resolution+printing.">Printing meets lithography: Soft approaches to high-resolution printing.</a> IBM Journal of Research and Development. 45 (5), 697-719 (2001).</li><li>Libioulle, L., Bietsch, A., Schmid, H., Michel, B., Delamarche, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contact-Inking+Stamps+for+Microcontact+Printing+of+Alkanethiols+on+Gold.">Contact-Inking Stamps for Microcontact Printing of Alkanethiols on Gold.</a> Langmuir. 15 (2), 300-304 (1999).</li><li>Sharpe, R. B. A., Burdinski, D., Huskens, J., Zandvliet, H. J. W., Reinhoudt, D. N., Poelsema, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spreading+of+16-Mercaptohexadecanoic+Acid+in+Microcontact+Printing.">Spreading of 16-Mercaptohexadecanoic Acid in Microcontact Printing.</a> Langmuir. 20 (20), 8646-8651 (2004).</li><li>Workman, R. K., Manne, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+Transfer+and+Transport+in+Noncovalent+Microcontact+Printing.">Molecular Transfer and Transport in Noncovalent Microcontact Printing.</a> Langmuir. 20 (3), 805-815 (2004).</li><li>Li, X. -. M., Peter, M., Huskens, J., Reinhoudt, D. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Catalytic+Microcontact+Printing+without+Ink.">Catalytic Microcontact Printing without Ink.</a> Nano Lett. 3 (10), 1449-1453 (2003).</li><li>Shestopalov, A. A., Clark, R. L., Toone, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inkless+Microcontact+Printing+on+Self-Assembled+Monolayers+of+Fmoc-Protected+Aminothiols.">Inkless Microcontact Printing on Self-Assembled Monolayers of Fmoc-Protected Aminothiols.</a> J. Am. Chem. Soc. 129 (145), 13818-13819 (2007).</li><li>Shestopalov, A. A., Clark, R. L., Toone, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Catalytic+Microcontact+Printing+on+Chemically+Functionalized+H-Terminated+Silicon.">Catalytic Microcontact Printing on Chemically Functionalized H-Terminated Silicon.</a> Langmuir. 26 (3), 1449-1451 (2010).</li><li>Shestopalov, A. A., Clark, R. L., Toone, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inkless+Microcontact+Printing+on+SAMs+of+Boc-+and+TBS-Protected+Thiols.">Inkless Microcontact Printing on SAMs of Boc- and TBS-Protected Thiols.</a> Nano Lett. 10 (1), 43-46 (2010).</li><li>Snyder, P. W., Johannes, M. S., Vogen, B. N., Clark, R. L., Toone, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biocatalytic+Microcontact+Printing.">Biocatalytic Microcontact Printing.</a> J. Org. Chem. 72 (19), 7459-7461 (2007).</li><li>Morris, C. J., Shestopalov, A. A., Gold, B. H., Clark, R. L., Toone, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patterning+NHS-Terminated+SAMs+on+Germanium.">Patterning NHS-Terminated SAMs on Germanium.</a> Langmuir. 27 (10), 6486-6489 (2011).</li><li>Shestopalov, A. A., Morris, C. J., Vogen, B. N., Hoertz, A., Clark, R. L., Toone, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Soft-Lithographic+Approach+to+Functionalization+and+Nanopatterning+Oxide-Free+Silicon.">Soft-Lithographic Approach to Functionalization and Nanopatterning Oxide-Free Silicon.</a> Langmuir. 27 (10), 6478-6485 (2011).</li></ol>

我々は、開示することは何もない

提示プロトコルは普遍的に単純な秩序化単分子膜をサポートできる任意の基材に適用することができるフォームまたはinklessマイクロコンタクトプリンティングのです。この方法では、スタンプが固定化触媒は、対応する官能基を有する表面にパターンを転送します。プロセスがタイムスタンプから表面へのインキ転移性に依存しないため、伝統的な反応性μCPの拡散解像度の制限は、ナノスケールのオブジェクトのルーチン製造を可能にして、排除される。主な高秩序の分子システムの導入は、酸化による損傷からの基盤となる半導体の完全な保護を提供します。それと同時に、方法は二次的反応性被覆層を利用することで、煩雑な反応性基の固定化をサポートし、一緒にシステ​​ムの保護と機能化を両立。 技術は化学的に不活性primarを可能にする安定した炭素表面結合の形成から始まり、酸化物形成への効果的なバリアとして機能するyの単層。二次反応上層の形成は、化学的および生物学的部分のさまざまな接続点として機能する端末のNHS官能基を提供します。この安定した二層の分子システムは、その後、私たちの触媒μCPのアプローチを用いてパターニングされる。本研究で紹介するアプローチは、有機と生物学的材料の広範なパターニング半導体基板の一般的な方法を提供しています。高価、複雑な計装なしでパターニングされた有機半導体のインターフェースを作成する機能は、エレクトロニクス、ナノテクノロジー、生化学、生物物理学などの分野で数多くの機会を提供しています。

<table border="1"><tbody><tr><th>試薬の名前</th><th>会社/モデル</th></tr><tr><td> XPSスペクトロメータ</td><td> KRATOS軸ウルトラ</td></tr><tr><td>原子間力顕微鏡</td><td> Veeco社D3100 </td></tr><tr><td> SEM - FEGの顕微鏡</td><td> FEI XL30 </td></tr><tr><td>蛍光顕微鏡</td><td>ツァイスアクシオイメージャ</td></tr><tr><td> Heatblock </td><td> VWR </td></tr><tr><td>真空ポンプ</td><td> BOCエドワーズ</td></tr><tr><td>水精製システム</td><td>ミリポア</td></tr><tr><td> TESPシリコンプローブ</td><td> Veeco社</td></tr><tr><td>シリコン</td><td></td></tr><tr><td>圧力バイアル</td><td> Chemglass </td></tr><tr><td>真空マニホールド</td><td> Chemglass </td></tr><tr><td> UVランプ</td><td> UVP </td></tr><tr><td>材料のスタンプ</td><td>リファレンス20と18を参照してください。 </td></tr><tr><td>シリンジフィルターをPFTE </td><td> VWR </td></tr><tr><td>ナノストリップ</td><td> Cyantek </td></tr><tr><td>塩酸</td><td>シグマ</td></tr><tr><td>エタノール</td><td>シグマ</td></tr><tr><td>アセトン</td><td>シグマ</td></tr><tr><td> HF </td><td>シグマ</td></tr><tr><td>クロロベンゼン</td><td>シグマ</td></tr><tr><td> PCL5 </td><td>シグマ</td></tr><tr><td>プロペニル塩化マグネシウム</td><td>シグマ</td></tr><tr><td>オクチル塩化マグネシウム</td><td>シグマ</td></tr><tr><td>四塩化炭素</td><td>シグマ</td></tr><tr><td> Boc保護エチレンジアミン</td><td>シグマ</td></tr><tr><td> TFA </td><td>シグマ</td></tr><tr><td>ナトリウム2 - mercaptoethanesulfonate </td><td>シグマ</td></tr><tr><td>ジオキサン中の4N HCl溶液</td><td>シグマ</td></tr><tr><td>リジン- N、N -二酢酸</td><td>シグマ</td></tr><tr><td>のEt 3 N </td><td>シグマ</td></tr><tr><td> DMF </td><td>シグマ</td></tr><tr><td> NISO 4 </td><td>シグマ</td></tr><tr><td> NAP </td><td>シグマ</td></tr><tr><td> NaClを</td><td>シグマ</td></tr><tr><td>イミダゾール</td><td>シグマ</td></tr><tr><td> PBS </TD&gt; <td>シグマ</td></tr></tbody></table>

soft lithographic functionalization and patterning oxide-free silicon and germanium

ハイブリッド電子機器の発展は、効率的な電子輸送を可能にし、酸化分解から、基礎となる基板を保護するための安定したインタフェースを介して（バイオ）、有機材料と無機半導体の統合の大部分が依存しています。 IV族半導体は、効果的に有機相と水両方のソリューションを通さない障壁として機能する単純なアルキル鎖から構成される高秩序の自己組織化単分子層（SAM）を使用して保護することができます。単純なアルキルSAMのは、しかし、不活性および伝統的なパターニング技術に従順ではありません。半導体上の有機分子システムを固定化するための動機は、光学、電子、および機械的な機能だけでなく、化学的および生物学的活性を提供することができる表面に新しい機能を付与することです。 マイクロコンタクトプリント（μCP）は、無数の表面のパターニングのSAM用ソフトリソグラフィ技術である。1-10そのあほにもかかわらずlicityと汎用性は、アプローチは、主に貴金属の表面に限定されており、よくそのような酸化物を含まないシリコンやゲルマニウムのような技術的に重要な基材へのパターン転写用に開発されていない。この技術は、基板にエラストマーからパターンを転写するインクの拡散に依存しているので更に、、このような伝統的な印刷の解像度は、本質的に近い1μmの10から16に制限されています従来の印刷とは対照的に、inklessμの CPのパターニングは、表面固定化基板とスタンプ結合型触媒との間の特定の反応に依存しています。技術が拡散SAMの形成に依存しないため、それが大幅patternable表面の多様性を拡大する。さらに、inklessテクニックは非常に小さい（&lt;200 nm）の機能の複製を容易にする、分子拡散によって課されたフィーチャサイズの制限がなくなります。17-23しかしながら、今までは、inklessμの CPは、主に劣化から基本的な表面を保護していないのパターニングが比較的不規則な分子システムのために使用されています。 ここでは、反応性有機単分子膜とシリコンとゲルマニウムの不動態化パターニングするためのシンプルで信頼性の高いハイスループット法を報告し、小分子とタンパク質の両方でパターン化された基質の選択的官能基化を示しています。技術は、酸化物を含まないシリコンとゲルマニウム上に予め形成されたNHS -反応性二層システムを利用しています。 NHSの部分は、NHS -活性化し、遊離カルボン酸の化学的に異なるパターンを生成するスルホン酸変性アクリルスタンプ付きパターン固有の方法で加水分解される。多くのμの CP技術の解決への重要な制限は、高忠実度の転送に必要な機械的な剛性が不足しているPDMS材料を使用することです。この制限を緩和するために我々は、ポリウレタンアクリレートポリマー、可能な比較的硬い材料を用い簡単に別の有機部分で官能。私たちのパターニングのアプローチは完全に、化学的酸化からシリコンとゲルマニウムの両方を保護するパターン化された特徴の形状と大きさを精密に制御を提供し、さらに有機的および生物学的分子と官能することができる化学的に判別パターンへの準備ができてアクセスすることができます。アプローチは、一般および他の技術的に関連の表面に適用可能です。

Watch this Scientific Journal Video about Soft Lithographic Functionalization and Patterning Oxide-free Silicon and Germanium at JoVE.com

Soft Lithographic Functionalization and Patterning Oxide-free Silicon and Germanium

ここでは、反応性有機単分子膜とパターニング酸化物フリーシリコンとゲルマニウムのためのシンプルな方法を説明し、小分子とタンパク質とのパターン化された基質の官能基を示す。アプローチは完全に、化学的酸化から表面を保護する機能の形態を正確に制御を提供し、化学的に判別パターンにすぐにアクセスを提供します。

ソフトリソグラフィ機能化およびパターニング酸化物フリーシリコンとゲルマニウム

The development of hybrid electronic devices relies in large part on the integration of (bio)organic materials and inorganic semiconductors through a ...

soft-lithographic-functionalization-patterning-oxide-free-silicon

Research

JoVE Journal

Bioengineering

14.6K ビュー.  Duke University. ここでは、反応性有機単分子膜とパターニング酸化物フリーシリコンとゲルマニウムのためのシンプルな方法を説明し、小分子とタンパク質とのパターン化された基質の官能基を示す。アプローチは完全に、化学的酸化から表面を保護する機能の形態を正確に制御を提供し、化学的に判別パターンにすぐにアクセスを提供します。

ビデオ: Soft Lithographic Functionalization and Patterning Oxide-free Silicon and Germanium

Plasma Lithography Surface Patterning for Creation of Cell Networks

Systematic manipulation of a cell microenvironment with micro- and nanoscale resolution is often required for deciphering various cellular and molecular phenomena. To address this requirement, we have developed a plasma lithography technique to manipulate the cellular microenvironment by creating a patterned surface with feature sizes ranging from 100 nm to millimeters. The goal of this technique is to be able to study, in a controlled way, the behaviors of individual cells as well as groups of cells and their interactions.
This plasma lithography method is based on selective modification of the surface chemistry on a substrate by means of shielding the contact of low-temperature plasma with a physical mold. This selective shielding leaves a chemical pattern which can guide cell attachment and movement. This pattern, or surface template, can then be used to create networks of cells whose structure can mimic that found in nature and produces a controllable environment for experimental investigations. The technique is well suited to studying biological phenomenon as it produces stable surface patterns on transparent polymeric substrates in a biocompatible manner. The surface patterns last for weeks to months and can thus guide interaction with cells for long time periods which facilitates the study of long-term cellular processes, such as differentiation and adaption. The modification to the surface is primarily chemical in nature and thus does not introduce topographical or physical interference for interpretation of results. It also does not involve any harsh or toxic substances to achieve patterning and is compatible for tissue culture. Furthermore, it can be applied to modify various types of polymeric substrates, which due to the ability to tune their properties are ideal for and are widely used in biological applications. The resolution achievable is also beneficial, as isolation of specific processes such as migration, adhesion, or binding allows for discrete, clear observations at the single to multicell level.
This method has been employed to form diverse networks of different cell types for investigations involving migration, signaling, tissue formation, and the behavior and interactions of neurons arraigned in a network.

A versatile plasma lithography technique has been developed to generate stable surface patterns for guiding cellular attachment. This technique can be applied to create cell networks including those that mimic natural tissues and has been used for studying several, distinct cell types.

細胞ネットワークの作成のためのプラズマのリソグラフィ表面パターニング

Systematic manipulation of a cell microenvironment with micro- and nanoscale resolution is often required for deciphering various cellular and molecular ...

生物工学

High-throughput Synthesis of Carbohydrates and Functionalization of Polyanhydride Nanoparticles

Transdisciplinary approaches involving areas such as material design, nanotechnology, chemistry, and immunology have to be utilized to rationally design efficacious vaccines carriers. Nanoparticle-based platforms can prolong the persistence of vaccine antigens, which could improve vaccine immunogenicity1. Several biodegradable polymers have been studied as vaccine delivery vehicles1; in particular, polyanhydride particles have demonstrated the ability to provide sustained release of stable protein antigens and to activate antigen presenting cells and modulate immune responses2-12.
The molecular design of these vaccine carriers needs to integrate the rational selection of polymer properties as well as the incorporation of appropriate targeting agents. High throughput automated fabrication of targeting ligands and functionalized particles is a powerful tool that will enhance the ability to study a wide range of properties and will lead to the design of reproducible vaccine delivery devices.
The addition of targeting ligands capable of being recognized by specific receptors on immune cells has been shown to modulate and tailor immune responses10,11,13 C-type lectin receptors (CLRs) are pattern recognition receptors (PRRs) that recognize carbohydrates present on the surface of pathogens. The stimulation of immune cells via CLRs allows for enhanced internalization of antigen and subsequent presentation for further T cell activation14,15. Therefore, carbohydrate molecules play an important role in the study of immune responses; however, the use of these biomolecules often suffers from the lack of availability of structurally well-defined and pure carbohydrates. An automation platform based on iterative solution-phase reactions can enable rapid and controlled synthesis of these synthetically challenging molecules using significantly lower building block quantities than traditional solid-phase methods16,17.
Herein we report a protocol for the automated solution-phase synthesis of oligosaccharides such as mannose-based targeting ligands with fluorous solid-phase extraction for intermediate purification. After development of automated methods to make the carbohydrate-based targeting agent, we describe methods for their attachment on the surface of polyanhydride nanoparticles employing an automated robotic set up operated by LabVIEW as previously described10. Surface functionalization with carbohydrates has shown efficacy in targeting CLRs10,11 and increasing the throughput of the fabrication method to unearth the complexities associated with a multi-parametric system will be of great value (Figure 1a).

In this article, a high throughput method is presented for the synthesis of oligosaccharides and their attachment to the surface of polyanhydride nanoparticles for further use in targeting specific receptors on antigen presenting cells.

無水物ナノ粒子のハイスループット糖鎖の合成と機能化

Transdisciplinary approaches involving areas such as material design, nanotechnology, chemistry, and immunology have to be utilized to rationally design ...

Visualization of Cortex Organization and Dynamics in Microorganisms, using Total Internal Reflection Fluorescence Microscopy

TIRF microscopy has emerged as a powerful imaging technology to study spatio-temporal dynamics of fluorescent molecules in vitro and in living cells1. The optical phenomenon of total internal reflection occurs when light passes from a medium with high refractive index into a medium with low refractive index at an angle larger than a characteristic critical angle (i.e. closer to being parallel with the boundary). Although all light is reflected back under such conditions, an evanescent wave is created that propagates across and along the boundary, which decays exponentially with distance, and only penetrates sample areas that are 100-200 nm near the interface2. In addition to providing superior axial resolution, the reduced excitation of out of focus fluorophores creates a very high signal to noise ratios and minimizes damaging effects of photobleaching2,3. Being a widefield technique, TIRF also allows faster image acquisition than most scanning based confocal setups.
At first glance, the low penetration depth of TIRF seems to be incompatible with imaging of bacterial and fungal cells, which are often surrounded by thick cell walls. On the contrary, we have found that the cell walls of yeast and bacterial cells actually improve the usability of TIRF and increase the range of observable structures4-8. Many cellular processes can therefore be directly accessed by TIRF in small, single-cell microorganisms, which often offer powerful genetic manipulation techniques. This allows us to perform in vivo biochemistry experiments, where kinetics of protein interactions and activities can be directly assessed in living cells.
We describe here the individual steps required to obtain high quality TIRF images for Saccharomyces cerevisiae or Bacillus subtilis cells. We point out various problems that can affect TIRF visualization of fluorescent probes in cells and illustrate the procedure with several application examples. Finally, we demonstrate how TIRF images can be further improved using established image restoration techniques.

Total Internal Reflection Fluorescence (TIRF) microscopy is a powerful approach to observe structures close to the cell surface at high contrast and temporal resolution. We demonstrate how TIRF can be employed to study protein dynamics at the cortex of cell wall-enclosed bacterial and fungal cells.

全内部反射蛍光顕微鏡を用いた微生物のCortex組織とダイナミクスの可視化、

TIRF microscopy has emerged as  a powerful imaging technology to study spatio-temporal dynamics of fluorescent molecules  in vitro and in living cells1. ...

Light-mediated Formation and Patterning of Hydrogels for Cell Culture Applications

Click chemistries have been investigated for use in numerous biomaterials applications, including drug delivery, tissue engineering, and cell culture. In particular, light-mediated click reactions, such as photoinitiated thiol&#8722;ene and thiol&#8722;yne reactions, afford spatiotemporal control over material properties and allow the design of systems with a high degree of user-directed property control. Fabrication and modification of hydrogel-based biomaterials using the precision afforded by light and the versatility offered by these thiol&#8722;X photoclick chemistries are of growing interest, particularly for the culture of cells within well-defined, biomimetic microenvironments. Here, we describe methods for the photoencapsulation of cells and subsequent photopatterning of biochemical cues within hydrogel matrices using versatile and modular building blocks polymerized by a thiol&#8722;ene photoclick reaction. Specifically, an approach is presented for constructing hydrogels from allyloxycarbonyl (Alloc)-functionalized peptide crosslinks and pendant peptide moieties and thiol-functionalized poly(ethylene glycol) (PEG) that rapidly polymerize in the presence of lithium acylphosphinate photoinitiator and cytocompatible doses of long wavelength ultraviolet (UV) light. Facile techniques to visualize photopatterning and quantify the concentration of peptides added are described. Additionally, methods are established for encapsulating cells, specifically human mesenchymal stem cells, and determining their viability and activity. While the formation and initial patterning of thiol-alloc hydrogels are shown here, these techniques broadly may be applied to a number of other light and radical-initiated material systems (e.g., thiol-norbornene, thiol-acrylate) to generate patterned substrates.

We describe a sequential process for light-mediated formation and subsequent biochemical patterning of synthetic hydrogel matrices for three-dimensional cell culture applications. The construction and modification of hydrogels with cytocompatible photoclick chemistry is demonstrated. Additionally, facile techniques to quantify and observe patterns and determine cell viability within these hydrogels are presented.

細胞培養アプリケーションのためのヒドロゲルの光媒介形成とパターニング

Click chemistries have been investigated for use in numerous biomaterials applications, including drug delivery, tissue engineering, and cell culture. In ...

A Versatile Method of Patterning Proteins and Cells

Substrate and cell patterning techniques are widely used in cell biology to study cell-to-cell and cell-to-substrate interactions. Conventional patterning techniques work well only with simple shapes, small areas and selected bio-materials. This article describes a method to distribute cell suspensions as well as substrate solutions into complex, long, closed (dead-end) polydimethylsiloxane (PDMS) microchannels using negative pressure. This method enables researchers to pattern multiple substrates including fibronectin, collagen, antibodies (Sal-1), poly-D-lysine (PDL), and laminin. Patterning of substrates allows one to indirectly pattern a variety of cells. We have tested C2C12 myoblasts, the PC12 neuronal cell line, embryonic rat cortical neurons, and amphibian retinal neurons. In addition, we demonstrate that this technique can directly pattern fibroblasts in microfluidic channels via brief application of a low vacuum on cell suspensions. The low vacuum does not significantly decrease cell viability as shown by cell viability assays. Modifications are discussed for application of the method to different cell and substrate types. This technique allows researchers to pattern cells and proteins in specific patterns without the need for exotic materials or equipment and can be done in any laboratory with a vacuum.

This report describes a simple, easy to perform technique, using low pressure vacuum, to fill microfluidic channels with cells and substrates for biological research.

パターニングタンパク質および細胞の多彩な方法

Substrate and cell patterning techniques are widely used in cell biology to study cell-to-cell and cell-to-substrate interactions. Conventional patterning ...

Soft Lithographic Procedure for Producing Plastic Microfluidic Devices with View-ports Transparent to Visible and Infrared Light

Infrared (IR) spectro-microscopy of living biological samples is hampered by the absorption of water in the mid-IR range and by the lack of suitable microfluidic devices. Here, a protocol for the fabrication of plastic microfluidic devices is demonstrated, where soft lithographic techniques are used to embed transparent Calcium Fluoride (CaF2) view-ports in connection with observation chamber(s). The method is based on a replica casting approach, where a polydimethylsiloxane (PDMS) mold is produced through standard lithographic procedures and then used as the template to produce a plastic device. The plastic device features ultraviolet/visible/infrared (UV/Vis/IR) -transparent windows made of CaF2 to allow for direct observation with visible and IR light. The advantages of the proposed method include: a reduced need for accessing a clean room micro-fabrication facility, multiple view-ports, an easy and versatile connection to an external pumping system through the plastic body, flexibility of the design, e.g., open/closed channels configuration, and the possibility to add sophisticated features such as nanoporous membranes.

A protocol for the fabrication of plastic microfluidic devices with transparent view-ports for visible and infrared light imaging is described.

ビュー ポートの可視光と赤外線を透明なプラスチックのマイクロ流体デバイスを製造するためソフト平版のプロシージャ

Infrared (IR) spectro-microscopy of living biological samples is hampered by the absorption of water in the mid-IR range and by the lack of suitable ...

Designing Porous Silicon Films as Carriers of Nerve Growth Factor

Despite the great potential of NGF for treating neurodegenerative diseases, its therapeutic administration represents a significant challenge as the protein does not cross the blood-brain barrier, owing to its chemical properties, and thus requires long-term delivery to the brain to have a biological effect. This work describes fabrication of nanostructured PSi films as degradable carriers of NGF for sustained delivery of this sensitive protein. The PSi carriers are specifically tailored to obtain high loading efficacy and continuous release of NGF for a period of four weeks, while preserving its biological activity. The behavior of the NGF-PSi carriers as a NGF delivery system is investigated in vitro by examining their capability to induce neuronal differentiation and outgrowth of PC12 cells and dissociated DRG neurons. Cell viability in the presence of neat and NGF-loaded PSi carriers is evaluated. The bioactivity of NGF released from the PSi carriers is compared to the conventional treatment of repetitive free NGF administrations. PC12 cell differentiation is analyzed and characterized by the measurement of three different morphological parameters of differentiated cells; (i) the number of neurites extracting from the soma (ii) the total neurites' length and (iii) the number of branching points. PC12 cells treated with the NGF-PSi carriers demonstrate a profound differentiation throughout the release period. Furthermore, DRG neuronal cells cultured with the NGF-PSi carriers show an extensive neurite initiation, similar to neurons treated with repetitive free NGF administrations. The studied tunable carriers demonstrate the long-term implants for NGF release with a therapeutic potential for neurodegenerative diseases.

Here, we present a protocol to design and fabricate nanostructured porous silicon (PSi) films as degradable carriers for the nerve growth factor (NGF). Neuronal differentiation and outgrowth of PC12 cells and mice dorsal root ganglion (DRG) neurons are characterized upon treatment with the NGF-loaded PSi carriers.

神経成長因子のキャリアとしてポーラス シリコン膜の設計

Despite the great potential of NGF for treating neurodegenerative diseases, its therapeutic administration represents a significant challenge as the ...

Bioinspired Soft Robot with Incorporated Microelectrodes

Bioinspired soft robotic systems that mimic living organisms using engineered muscle tissue and biomaterials are revolutionizing the current biorobotics paradigm, especially in biomedical research. Recreating artificial life-like actuation dynamics is crucial for a soft-robotic system. However, the precise control and tuning of actuation behavior still represents one of the main challenges of modern soft robotic systems. This method describes a low-cost, highly scalable, and easy-to-use procedure to fabricate an electrically controllable soft robot with life-like movements that is activated and controlled by the contraction of cardiac muscle tissue on a micropatterned sting ray-like hydrogel scaffold. The use of soft photolithography methods makes it possible to successfully integrate multiple components in the soft robotic system, including micropatterned hydrogel-based scaffolds with carbon nanotubes (CNTs) embedded gelatin methacryloyl (CNT-GelMA), poly(ethylene glycol) diacrylate (PEGDA), flexible gold (Au) microelectrodes, and cardiac muscle tissue. In particular, the hydrogels alignment and micropattern are designed to mimic the muscle and cartilage structure of the sting ray. The electrically conductive CNT-GelMA hydrogel acts as a cell scaffold that improves the maturation and contraction behavior of cardiomyocytes, while the mechanically robust PEGDA hydrogel provides structural cartilage-like support to the whole soft robot. To overcome the hard and brittle nature of metal-based microelectrodes, we designed a serpentine pattern that has high flexibility and can avoid hampering the beating dynamics of cardiomyocytes. The incorporated flexible Au microelectrodes provide electrical stimulation across the soft robot, making it easier to control the contraction behavior of cardiac tissue.

A bioinspired scaffold is fabricated by a soft photolithography technique using mechanically robust and electrically conductive hydrogels. The micropatterned hydrogels provide directional cardiomyocyte cell alignment, resulting in a tailored direction of actuation. Flexible microelectrodes are also integrated into the scaffold to bring electrical controllability for a self-actuating cardiac tissue.

生体にインスパイアされたマイクロ電極を用いたソフトロボット

Bioinspired soft robotic systems that mimic living organisms using engineered muscle tissue and biomaterials are revolutionizing the current biorobotics ...

Silicon Nanowires and Optical Stimulation for Investigations of Intra- and Intercellular Electrical Coupling

Myofibroblasts can spontaneously internalize silicon nanowires (SiNWs), making them an attractive target for bioelectronic applications. These cell-silicon hybrids offer leadless optical modulation capabilities with minimal perturbation to normal cell behavior. The optical capabilities are obtained by the photothermal and photoelectric properties of SiNWs. These hybrids can be harvested using standard tissue culture techniques and then applied to different biological scenarios. We demonstrate here how these hybrids can be used to study the electrical coupling of cardiac cells and compare how myofibroblasts couple to one another or to cardiomyocytes. This process can be accomplished without special equipment beyond a fluorescent microscope with coupled laser line. Also shown is the use of a custom-built MATLAB routine that allows the quantification of calcium propagation within and between the different cells in the culture. Myofibroblasts are shown to have a slower electrical response than that of cardiomyocytes. Moreover, the myofibroblast intercellular propagation shows slightly slower, though comparable velocities to their intracellular velocities, suggesting passive propagation through gap junctions or nanotubes. This technique is highly adaptable and can be easily applied to other cellular arenas, for in vitro as well as in vivo or ex vivo investigations.

This protocol describes the use of silicon nanowires for intracellular optical bio-modulation of cell in a simple and easy to perform method. The technique is highly adaptable to diverse cell types and can be used for in vitro as well as in vivo applications.

シリコンナノワイヤと細胞間電気結合の研究のための光刺激

Myofibroblasts can spontaneously internalize silicon nanowires (SiNWs), making them an attractive target for bioelectronic applications. These ...

Simple, Affordable, and Modular Patterning of Cells using DNA

The relative positioning of cells is a key feature of the microenvironment that organizes cell-cell interactions. To study the interactions between cells of the same or different type, micropatterning techniques have proved useful. DNA Programmed Assembly of Cells (DPAC) is a micropatterning technique that targets the adhesion of cells to a substrate or other cells using DNA hybridization. The most basic operations in DPAC begin with decorating cell membranes with lipid-modified oligonucleotides, then flowing them over a substrate that has been patterned with complementary DNA sequences. Cells adhere selectively to the substrate only where they find a complementary DNA sequence. Non-adherent cells are washed away, revealing a pattern of adherent cells. Additional operations include further rounds of cell-substrate or cell-cell adhesion, as well as transferring the patterns formed by DPAC to an embedding hydrogel for long-term culture. Previously, methods for patterning oligonucleotides on surfaces and decorating cells with DNA sequences required specialized equipment and custom DNA synthesis, respectively. We report an updated version of the protocol, utilizing an inexpensive benchtop photolithography setup and commercially available cholesterol modified oligonucleotides (CMOs) deployed using a modular format. CMO-labeled cells adhere with high efficiency to DNA-patterned substrates. This approach can be used to pattern multiple cell types at once with high precision and to create arrays of microtissues embedded within an extracellular matrix. Advantages of this method include its high resolution, ability to embed cells into a three-dimensional microenvironment without disrupting the micropattern, and flexibility in patterning any cell type.

Here we present a protocol to micropattern cells at single-cell resolution using DNA-programmed adhesion. This protocol uses a benchtop photolithography platform to create patterns of DNA oligonucleotides on a glass slide and then labels cell membranes with commercially available complementary oligonucleotides. Hybridization of the oligos results in programmed cell adhesion.

DNAを用いた細胞のシンプル、手頃な価格、モジュール式パターニング

The relative positioning of cells is a key feature of the microenvironment that organizes cell-cell interactions. To study the interactions between cells ...