この材料は、米国エネルギー省、材料科学とエンジニアリング部門（受賞番号DEFG - 02 - 07ER46471）とライト材料DOEエネルギー研究センターエネルギー変換の相互作用（によってサポートされている仕事に基づいています。賞第DE - SC0001293 ）、およびフレデリックサイツ材料研究所（FSMRL）内の材料の微量分析のためのセンターへのアクセスから恩恵。

1。はじめ<ol><li>本論文では、1D、2D、および直接書き込みアセンブリを介して導電性微小電極の3Dプリントを示しています。 </li><li>直接書き込みアセンブリは、微細なノズルを介して濃縮されたインキの堆積により、1Dから3Dプリント構造を構築する方法です。 </li><li>私たちのシステムはコンピュータ制御の3軸移動ステージ、インクタンクとノズル、およびイメージングのための10倍望遠レンズ（図1）で構成されています。 </li><li>直接書き込みアセンブリは、濃縮されたインクは、その直径0.1〜250μmである（図2）の範囲の円筒形のノズルを通して押出されたフィラメント状の印刷のアプローチです。特に、粘弾性のインクの特性により、直接書き込みアセンブリは、自己支持スパニング機能（図3）を可能にします。日付、セラミック6,7で構成されるものを含むインクの広い範囲、、有機8-10、メタリック11-15、高分子16,17、およびゾル-ゲル18,19の材料にされているこの印刷手法（図4）のために開発。 </li></ol> 2。高濃度の銀ナノ粒子インクの調製<ol><li>銀ナノ粒子インクは、最初の50 gの水と40グラムジエタノールアミン（ビデオ2.1）の混合物にポリ5,000〜50,000の分子量（アクリル酸）のブレンドを溶解することにより調製されています。 </li><li>ポリマーは、銀ナノ粒子のサイズを制御するためのキャッピング剤として作用する。 </li><li>次に、硝酸銀の水溶液は、ポリマー溶液に注入されます。添加後、淡黄色透明溶液（ビデオ2.2）得られる。 </li><li>室温で24時間攪拌した後、解決策は、透過型電子顕微鏡によって決定される5nm以下の直径の銀ナノ粒子の形成と一致する赤褐色の色を（ビデオ2.3）、開発しています。 </li><li>次に、溶液を65℃でさらに粒子成長（ビデオ2.4）のための2時間、水浴中で超音波処理しています。 </li><li> AfteR超音波処理、解決策を500mlビーカーに移し、室温に冷却される。その後、エタノール300mlを30ml /分の速度で滴定する。エタノールは、ポリ（アクリル酸）キャッピング剤の貧溶媒であるので、粒子が急速に凝固し、解決策（ビデオ2.5）から沈殿。 </li><li>上清をデカントした後、沈殿物を遠心チューブに回収され、20分（ビデオ2.6）9000 rpmで遠心分離。 </li><li>このステップの後、〜85重量％の固形物負荷で高濃度の銀ナノ粒子インクは（ビデオ2.7）得られる。 </li><li>インクの粘度と弾性率をさらにコントロールは、均質化に続いて、希釈することによって達成することができます。例えば、エチレングリコールなどの湿潤剤溶液は、インクに追加することができますし、シンキー均質化のミキサーを使用して3分間2000rpmでホモジナイズした。このプロセスの後に、マゼンタの色〜青系の均一なインクは、（ビデオ2.8）得られる。 </li><li> TEM像を示しています銀は、この合成手順（図5_left）によって得られたナノ粒子。粒子は50〜50 nmの粒径分布を持つ20nmの平均直径を有する。印刷された構造は、その導電性を高めるためにポストアニールが必要です。未満で30分間250℃でアニーリングした後、銀ナノ粒子は、電気抵抗率が10 -5Ω•CM（図5_bottomの右側）に近づくと導電性微小電極を形成する。アニーリング温度の関数として印刷された銀の微小電極の微細構造発達を図5_topの右側に表示されます。温度は150℃から550℃に増加すると、微小電極は、〜30％の総体積収縮で高密度化を受ける。11 </li><li>強く固形負荷に依存するインクのレオロジーは、、その印刷適性を決定する。増加する固形のロードとインクの粘度が増加する（図6）。ための重要な側面の拡散、concentrで低粘度の結果との希薄なインク70〜85重量％の範囲の固体のロードとatedインクは、平面とスパニングインキフィラメントの印刷に必要とされています。 </li><li>増加する固形のロードとインク弾性率が増加する（図7）。線形粘弾性領域で、弾性率は60〜75重量％を増加させる固体ロードとして大きさのほぼ3桁の上昇。 Paの2000年の最小弾性係数は、機能を自立またはスパニング生成するために必要です。 </li></ol> 3。直接書き込みアセンブリ<ol><li>直接書き込みアセンブリは、最初にシリンジバレルにインクをロードすることにより行われる。堆積ノズルを取り付けた後、インクロードされたシリンジバレルは、3軸の印刷段階（ビデオ3.1）の上にマウントされています。 </li><li>コンピュータプログラムを使用して、、線形平面、および複雑な三次元構造を含む任意のデザインを、簡単に（ビデオ3.2）が生成することができます。 </li><li>次に、ノズルの高さは10倍ズーム（ビデオ3.3）と望遠鏡のレンズの助けを借りて調整されます。 </l私&gt; <li>エアー駆動液体ディスペンシングシステムを用いて圧力を適用した後、インクが制御された印刷速度（ビデオ3.4）で基板上に堆積される。必要な圧力は、インクのレオロジー、ノズルの直径、および印刷速度が、20から500ミクロン/ sで10から100 psiから典型的な値の範囲に依存この印刷は、室温で空気中で行われます。この印刷手順を使用して、別のレイアウトやサイズのスケールにおける銀の微小電極の印刷が示されている。 </li><li>例えば、シリコンウエハー基板上に5μmのノズルによってパターン化さ100μmの中心間の行間隔を持つ導電性銀格子、の印刷は（ビデオ3.5）が示されている。 </li><li>さらに、このビデオでは、層ごとの印刷方法（ビデオ3.6）を用いて30μmのノズルによる高アスペクト比円筒形の構造を作成する方法を示します。 </li><li>さらに、2枚のガラス基板の間に銀の微小電極の全方位印刷は、1 mmの高さがディで相殺fferenceは30μmのノズルを（ビデオ3.7）を使用して示されています。 </li><li>完全にフリースタンディング、垂直に印刷された銀のmicrospikesは、Siウェハ基板（ビデオ3.8）に30μmのノズルで作成することができます。 </li><li>最後に、このビデオは10μmのノズルを（ビデオ3.9）を使用してスパニング銀の微小電極の直接の書き込みを示しています。印刷機能は、最小限の垂下や座屈と1センチメートルの距離をまたがることができます。 </li></ol> 4。代表的な結果： 我々は、高濃度の銀ナノ粒子インクを用意し、〜2の印刷解像度で、電子と光電子アプリケーションのために平面と3Dモチーフに印刷された導電性の機能を実証 - 30μmの。例えば、図8は展示この手法の印刷解像度を。 〜2μmの（1.4μm厚）の最小電極幅と印刷機能は、1μmのノズル11を使用して単一のパスで得られる。 図9に示すの透明導電性銀グリッド、柔軟性のあるポリイミド膜12上に5μmのノズルによってパターン化された。印刷されたグリッドの下にテキストがはっきりとわかります。これらの透明な銀のグリッドは、透明導電酸化物（TCO）の材料のための魅力的な選択肢かもしれない。 非平面基板上への等角印刷もこの方法で有効になっています。図10は、3次元電気的に小さなアンテナのコンフォーマル印刷を示しています。 100μmの金属のノズルはガラスの半球13の表面上に蛇行ラインパターンを印刷するために使用されます。このアプローチは、植込み型ウェアラブルアンテナ、電子機器、センサーなどいくつかのアプリケーションを見つけることができます。 三次元太陽光発電や発光ダイオードに銀電極をまたがるのアプリケーションは、（図11-14）が実証されています。 最初に、図11は、シリコン球殻の例です。 2 -μmのtで、この微妙な映画hicknessは、全方位印刷14による外部回路にワイヤボンディングすることができます。このメソッドは、デリケートなデバイスに非常に有利な最小限の接触圧を、使用しています。 次に、図12は、シリコンmicroribbonの要素が33μmのギャップ15によって分離されているシリコン太陽電池マイクロセルアレイのスパニングインターコネクトを出力する例を示しています。 銀は、ガリウム砒素ベースの各ピクセル（500 × 500 × 2.5μmの3）を離れて11200μmを間隔をあけている。4 × 4ピクセル、LEDアレイの相互接続次に、図13に示すよう下の画像は、単一のピクセルから6 Vの印加されたバイアス下で均一な赤色光を発光する、LEDアレイが表示されます。スパニング電極を印刷する機能は、支持または犠牲層（トップ画像）を使用せずに多層配線を可能にします。 複雑な3DのマイクのためのSEM画像を最終的なデモンストレーション、図14に示すように5μmのノズルで印刷roperiodic銀格子。 <img alt="図1" src="/files/ftp_upload/3189/3189fig1.jpg" /> 図1。ダイレクトインクジェット描画装置の光学像。 <img alt="図2" src="/files/ftp_upload/3189/3189fig2.jpg" /> 図2。フィラメント機能の直接インク描画。 <img alt="図3" src="/files/ftp_upload/3189/3189fig3.jpg" /> 図3。自立スパニング機能の直接インク描画。 <img alt="図4" src="/files/ftp_upload/3189/3189fig4.jpg" /> 図4直接インク描画用インクの設計。濃縮された粘弾性インクの広い範囲は、マイクロスケールの機能を有する平面と複雑な3次元構造の直接描画のために開発されている。 <img alt="図5" src="/files/ftp_upload/3189/3189fig5.jpg" /> 図5（左）銀ナノ粒子の透過型電子顕微鏡（TEM）像。 （右上）アニーリング温度の関数として、15μmのノズルでパターン化銀微小電極のSEM写真。 （右下）の温度と時間をアニーリングの関数として銀微小電極の電気抵抗率。 <img alt="図6" src="/files/ftp_upload/3189/3189fig6.jpg" /> 図6。固形物負荷の関数としての銀ナノ粒子インクの見かけ粘度（η）。 <img alt="図7" src="/files/ftp_upload/3189/3189fig7.jpg" /> 図7。固形物の荷重を変化させるの銀ナノ粒子インクのためのせん断応力の関数としてせん断弾性率（G&#39;）。 <img alt="図8" src="/files/ftp_upload/3189/3189fig8.jpg" /> 図8。パターン化銀微小電極の平面アレイのSEM写真1 -μmのノズル付きのSiウェハ上に。 <img alt="図9" src="/files/ftp_upload/3189/3189fig9.jpg" /> 図9。透明導電性銀グリッド（左）とラインのピッチ（右）の関数として印刷された格子のSEM像の光学像。 <img alt="図10" src="/files/ftp_upload/3189/3189fig10.jpg" /> 図10。半球ガラス基板上に電気的に小さなアンテナのコンフォーマル印刷中に取り込まれた光学像。 <img alt="図11" src="/files/ftp_upload/3189/3189fig11.jpg" /> 図11。細い（2 -μm）のシリコン球状の殻の上にまたがる銀微小電極の印刷中に得られた光学像。 <img alt="図12" src="/files/ftp_upload/3189/3189fig12.jpg" /> 図12ように、シリコンの上に印刷されたスパニング銀の微小電極のSEM像LARマイクロセル配列。 <img alt="図13" src="/files/ftp_upload/3189/3189fig13.jpg" /> 図13。SEM像（上）と銀微小電極によって相互接続された4 × 4のLEDチップの配列の光学像（下）。 <img alt="図14" src="/files/ftp_upload/3189/3189fig14.jpg" /> 図14 3D microperiodic銀の格子のSEM像。

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U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-resolution+electrohydrodynamic+jet+printing.">High-resolution electrohydrodynamic jet printing.</a> Nat. Mater. 6, 782-789 (2007).</li><li>Schirmer, N. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=On+ejecting+colloids+against+capillarity+from+sub-micrometer+openings:+On-demand+dielectrophoretic+nanoprinting.">On ejecting colloids against capillarity from sub-micrometer openings: On-demand dielectrophoretic nanoprinting.</a> Adv. Mater. 22, 4701-4705 (2010).</li><li>Park, J. -. U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=electrofield+liquid+jets+for+high-resolution+printing+of+charge.">electrofield liquid jets for high-resolution printing of charge.</a> Nano. Lett. 10, 584-591 (2010).</li><li>Ghosh, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct-write+assembly+of+micro-periodic+silk+fibroin+scaffolds+for+tissue+engineering+applications.">Direct-write assembly of micro-periodic silk fibroin scaffolds for tissue engineering applications.</a> Adv. Funct. Mater. 18, 1883-1889 (2008).</li><li>Barry, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct-write+assembly+of+3D+hydrogel+scaffolds+for+guided+cell+growth.">Direct-write assembly of 3D hydrogel scaffolds for guided cell growth.</a> Adv. Mater. 21, 2407-2410 (2009).</li><li>Shepherd, J. N. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=3D+microperiodic+hydrogel+scaffolds+for+robust+neuronal+cultures.">3D microperiodic hydrogel scaffolds for robust neuronal cultures.</a> Adv. Mater. 21, 47-54 (2011).</li><li>Wu, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct-write+assembly+of+biomimetic+microvascular+networks+for+efficient+fluid+transport.">Direct-write assembly of biomimetic microvascular networks for efficient fluid transport.</a> Soft. Matter. 6, 739-742 (2010).</li><li>Wu, W., DeConinck, A., Lewis, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Omnidirectional+Printing+of+3D+Microvascular+Networks.">Omnidirectional Printing of 3D Microvascular Networks.</a> Adv. Mater. 23, H178-H183 (2011).</li></ol>

利害の衝突は宣言されません。

このようなインクジェット印刷など、従来の液滴ベースの印刷のアプローチは、、希薄自然と使用するインクの低粘度のための低アスペクト比を持つ平面電極の製造に限定されています。最近、ディップペンナノリソグラフィー（DPN）20から22と、電子インクジェット23〜25は、パターンの導電機能するために使用されている。これらのルートはまた、希釈、低粘度のインクを採用。 Peartonと共同研究者は約0.5μm22の最大1600μmのs -1とし、線幅の書き込み速度で、市販の銀ナノ粒子インクを堆積させるためにDPNを使用。しかし、広い地域にわたって再現可能なパターンの製作は、このアプローチによって実証されていない。銀ナノ粒子インクも〜1.5μmの25の線幅を有する導電トレースを形成するための電子インクジェット印刷によって堆積されている。しかし、インクジェット印刷と同様に、不均一な印刷機能には、衛星滴形成と不均一なドロップDが原因で起こることが24,25を rying。 上記で示したように、濃縮された銀ナノ粒子インクの直接書き込みアセンブリは、フィラメントベースの印刷のアプローチを通じて、これらの限界を克服しています。この手法は、1D、2D、および3Dアーキテクチャの作成を可能に通過する、単一で高アスペクト比（H / W≈1.0）と導電性微小電極の製造を可能にします。印刷機能の大きさは、ノズルの直径、インクの固形分のロード、加圧力、および印刷速度に依存します。日付、小さいなどの導電トレースに〜2μmのは控えめな速度（&lt;2ミリメートルの-1）で1μmのノズルを使用してパターニングされているとして。インク組成物とノズルの形状を調整しては10cm s -1を超える最大印刷速度が可能です。しかし、微細なノズル（&lt;5μm）を用いての高速印刷が重要な課題である。 直接書き込みアセンブリのアプリケーションを実証するために、我々は導電性グリッド、ELを作製ectrically小型アンテナ、太陽電池、そして平面とスパニング印刷電極（図8-14）と発光ダイオード。特に、我々のアプローチは、金属構造物の作成に限定されるものではありません。このような絹フィブロイン、ハイドロゲルと逃亡中の有機インクに基づくものなど、他のインクの設計、使用して、我々は、直接書き込みアセンブリ26から30を経由して組織工学と細胞培養のための3Dスキャフォールドおよび微小血管ネットワークを構築した。 未来に向かって、多くの機会と課題があります。さらなる進歩は、新しいインクの設計、インクの流動ダイナミクスの優れたモデリング、および強化されたロボットと制御システムが必要です。高いスループットとナノスケールの分解能で3次元構造への1Dの大面積作製（&lt;100 nm）は重要な課題である。

<table border="1"><tr><td> 試薬の名前 </td><td> 会社 </td><td> カタログ番号 </td><td> コメント </td></tr><tr><td>ポリ（アクリル酸） </td><td> Polysciences、（株） </td><td> 06519 </td><td> MW 5000グラム/モル</td></tr><tr><td>ポリ（アクリル酸） </td><td> Polysciences、（株） </td><td> 00627 </td><td> MW5万グラム/モル</td></tr><tr><td>硝酸銀</td><td>シグマアルドリッチ</td><td> 209139 </td><td>シルバー源</td></tr><tr><td>ジエタノールアミン</td><td>シグマアルドリッチ</td><td> D8885 </td><td>溶剤/還元剤</td></tr><tr><td>エチレングリコール</td><td>シグマアルドリッチ</td><td> 102466 </td><td>保湿剤</td></tr><tr><td> Sonicater </td><td>フィッシャーサイエンティフィック</td><td> FS30H </td><td> - </td></tr><tr><td>遠心</td><td>ベックマンコールター</td><td>アヴァンティTM J - 25 I </td><td> - </td></tr><tr><td>ロボットステージ</td><td>エアロテック（株） </td><td> ABL 900010 </td><td> 3軸のモーション</td></tr><tr><td>注射筒</td><td> EFD社</td><td> 5109LBP - B </td><td> 3ミリリットル</td></tr><tr><td>ノズル</td><td> EFD社</td><td> - </td><td> ID = 0.1〜250μmの</td></tr><tr><td>ディスペンサー</td><td> EFD社</td><td> 800 </td><td>エア駆動</td></tr><tr><td>デザインソフトウェア</td><td>カスタム設計</td><td> - </td><td> Mingjie徐</td></tr></table>

planar and three-dimensional printing of conductive inks

プリンテッドエレクトロニクスは1-3柔軟なまたは多次元電子、光、および生物医学的装置を作成するために、低コスト、大面積の製造ルートに依存しています。本稿では、一（1D）、2（2D）、および伸縮柔軟性の形で導電性金属インク、およびスパニング微小電極の三次元（3D）印刷に焦点を当てる。 ダイレクトライトアセンブリ4,5は、微細なノズル（ - 250μm以下〜0.1）によって濃縮されたインキの堆積によって、単純な線から複雑な構造に至るまでの機能の製造を可能にする1対の3Dプリント技術です。この印刷方式は、コンピュータ制御の3軸移動ステージ、インクタンクとノズル、および視覚化のための10倍の望遠レンズで構成されています。インクジェット印刷とは異なり、液滴ベースのプロセスは、直接書き込みアセンブリのいずれか、または面外インキフィラメントの押出しを含む。印刷されたフィラメントは、通常、ノズルサイズに準拠しています。 Henceは、マイクロスケールの特徴（&lt;1μm）の大きな配列や多次元のアーキテクチャにパターニングし、組み立てることができる。 本稿では、まず直接書き込みアセンブリを経由して平面と3Dプリントのために高濃度の銀ナノ粒子インクを合成する。次に、多次元モチーフに微小電極を印刷するための標準プロトコルが示されています。最後に、電気的に小型のアンテナ、太陽電池、発光ダイオードのための印刷された微小電極のアプリケーションが強調表示されます。

Watch this Scientific Journal Video about Planar and Three-Dimensional Printing of Conductive Inks at JoVE.com

Planar and Three-Dimensional Printing of Conductive Inks

導電性金属インクの平面と立体印刷が説明されています。我々のアプローチは、マイクロスケールでの異常なレイアウトで印刷された電子、光、および生物医学的装置を製造するための新たな道を提供します。

導電性インクの平面および三次元印刷

Printed electronics rely on low-cost, large-area fabrication routes to create flexible or multidimensional electronic, optoelectronic, and biomedical ...

planar-and-three-dimensional-printing-of-conductive-inks

Research

JoVE Journal

Bioengineering

36.9K ビュー.  University of Illinois at Urbana-Champaign. 導電性金属インクの平面と立体印刷が説明されています。我々のアプローチは、マイクロスケールでの異常なレイアウトで印刷された電子、光、および生物医学的装置を製造するための新たな道を提供します。

ビデオ: Planar and Three-Dimensional Printing of Conductive Inks

Three-dimensional Optical-resolution Photoacoustic Microscopy

Optical microscopy, providing valuable insights at the cellular and organelle levels, has been widely recognized as an enabling biomedical technology. As the mainstays of in vivo three-dimensional (3-D) optical microscopy, single-/multi-photon fluorescence microscopy and optical coherence tomography (OCT) have demonstrated their extraordinary sensitivities to fluorescence and optical scattering contrasts, respectively. However, the optical absorption contrast of biological tissues, which encodes essential physiological/pathological information, has not yet been assessable. 
The emergence of biomedical photoacoustics has led to a new branch of optical microscopy optical-resolution photoacoustic microscopy (OR-PAM)1, where the optical irradiation is focused to the diffraction limit to achieve cellular1 or even subcellular2 level lateral resolution. As a valuable complement to existing optical microscopy technologies, OR-PAM brings in at least two novelties. First and most importantly, OR-PAM detects optical absorption contrasts with extraordinary sensitivity (i.e., 100%). Combining OR-PAM with fluorescence microscopy3 or with optical-scattering-based OCT4 (or with both) provides comprehensive optical properties of biological tissues. Second, OR-PAM encodes optical absorption into acoustic waves, in contrast to the pure optical processes in fluorescence microscopy and OCT, and provides background-free detection. The acoustic detection in OR-PAM mitigates the impacts of optical scattering on signal degradation and naturally eliminates possible interferences (i.e., crosstalks) between excitation and detection, which is a common problem in fluorescence microscopy due to the overlap between the excitation and fluorescence spectra. 
Unique for optical absorption imaging, OR-PAM has demonstrated broad biomedical applications since its invention, including, but not limited to, neurology5, 6, ophthalmology7, 8, vascular biology9, and dermatology10. In this video, we teach the system configuration and alignment of OR-PAM as well as the experimental procedures for in vivo functional microvascular imaging.

Optical-resolution photoacoustic microscopy (OR-PAM) is an emerging technology capable of imaging optical absorption contrasts in vivo with cellular resolution and sensitivity. Here, we provide a visualized instruction on the experimental protocols of OR-PAM, including system configuration, system alignment, typical in vivo experimental procedures, and functional imaging schemes.

三次元光学分解能光音響顕微鏡

Optical microscopy, providing valuable insights at the cellular and organelle levels, has been widely recognized as an enabling biomedical technology. As ...

生物工学

Alginate Hydrogels for Three-Dimensional Organ Culture of Ovaries and Oviducts

Ovarian cancer is the fifth leading cause of cancer deaths in women and has a 63% mortality rate in the United States1. The cell type of origin for ovarian cancers is still in question and might be either the ovarian surface epithelium (OSE) or the distal epithelium of the fallopian tube fimbriae2,3. Culturing the normal cells as a primary culture in vitro will enable scientists to model specific changes that might lead to ovarian cancer in the distinct epithelium, thereby definitively determining the cell type of origin. This will allow development of more accurate biomarkers, animal models with tissue-specific gene changes, and better prevention strategies targeted to this disease.
Maintaining normal cells in alginate hydrogels promotes short term in vitro culture of cells in their three-dimensional context and permits introduction of plasmid DNA, siRNA, and small molecules. By culturing organs in pieces that are derived from strategic cuts using a scalpel, several cultures from a single organ can be generated, increasing the number of experiments from a single animal. These cuts model aspects of ovulation leading to proliferation of the OSE, which is associated with ovarian cancer formation. Cell types such as the OSE that do not grow well on plastic surfaces can be cultured using this method and facilitate investigation into normal cellular processes or the earliest events in cancer formation4.
Alginate hydrogels can be used to support the growth of many types of tissues5. Alginate is a linear polysaccharide composed of repeating units of &beta;-D-mannuronic acid and &alpha;-L-guluronic acid that can be crosslinked with calcium ions, resulting in a gentle gelling action that does not damage tissues6,7. Like other three-dimensional cell culture matrices such as Matrigel, alginate provides mechanical support for tissues; however, proteins are not reactive with the alginate matrix, and therefore alginate functions as a synthetic extracellular matrix that does not initiate cell signaling5. The alginate hydrogel floats in standard cell culture medium and supports the architecture of the tissue growth in vitro.
A method is presented for the preparation, separation, and embedding of ovarian and oviductal organ pieces into alginate hydrogels, which can be maintained in culture for up to two weeks. The enzymatic release of cells for analysis of proteins and RNA samples from the organ culture is also described. Finally, the growth of primary cell types is possible without genetic immortalization from mice and permits investigators to use knockout and transgenic mice.

Culture of normal cells in their three-dimensional context represents an alternative method to study early events required for cellular transformation and tumorigenesis. This method is used to grow normal ovarian and oviductal cells to study early events in ovarian cancer formation.

卵巣と卵管の三次元器官培養用アルギン酸ゲル

Ovarian cancer is the fifth leading cause of cancer deaths in women and has a 63% mortality rate in the United States1. The cell type of origin for ...

Investigating the Three-dimensional Flow Separation Induced by a Model Vocal Fold Polyp

The fluid-structure energy exchange process for normal speech has been studied extensively, but it is not well understood for pathological conditions. Polyps and nodules, which are geometric abnormalities that form on the medial surface of the vocal folds, can disrupt vocal fold dynamics and thus can have devastating consequences on a patient&#39;s ability to communicate. Our laboratory has reported particle image velocimetry (PIV) measurements, within an investigation of a model polyp located on the medial surface of an in&#160;vitro driven vocal fold model, which show that such a geometric abnormality considerably disrupts the glottal jet behavior. This flow field adjustment is a likely reason for the severe degradation of the vocal quality in patients with polyps. A more complete understanding of the formation and propagation of vortical structures from a geometric protuberance, such as a vocal fold polyp, and the resulting influence on the aerodynamic loadings that drive the vocal fold dynamics, is necessary for advancing the treatment of this pathological condition. The present investigation concerns the three-dimensional flow separation induced by a wall-mounted prolate hemispheroid with a 2:1 aspect ratio in cross flow, i.e. a model vocal fold polyp, using an oil-film visualization technique. Unsteady, three-dimensional flow separation and its impact of the wall pressure loading are examined using skin friction line visualization and wall pressure measurements.

Vocal fold polyps can disrupt vocal fold dynamics and thus can have devastating consequences on a patient&#39;s ability to communicate. Three-dimensional flow separation induced by a wall-mounted model polyp and its impact on the wall pressure loading are examined using particle image velocimetry, skin friction line visualization, and wall pressure measurements.

モデルの声帯ポリープによる三次元流分離の調査

The fluid-structure energy exchange process for normal speech has been studied extensively, but it is not well understood for pathological conditions. ...

Human Cartilage Tissue Fabrication Using Three-dimensional Inkjet Printing Technology

Bioprinting, which is based on thermal inkjet printing, is one of the most attractive enabling technologies in the field of tissue engineering and regenerative medicine. With digital control&#160;cells, scaffolds, and growth factors can be precisely deposited to the desired two-dimensional (2D) and three-dimensional (3D) locations rapidly. Therefore, this technology is an ideal approach to fabricate tissues mimicking their native anatomic structures. In order to engineer cartilage with native zonal organization, extracellular matrix composition (ECM), and mechanical properties, we developed a bioprinting platform using a commercial inkjet printer with simultaneous photopolymerization capable for 3D cartilage tissue engineering. Human chondrocytes suspended in poly(ethylene glycol) diacrylate (PEGDA) were printed for 3D neocartilage construction via layer-by-layer assembly. The printed cells were fixed at their original deposited positions, supported by the surrounding scaffold in simultaneous photopolymerization. The mechanical properties of the printed tissue were similar to the native cartilage. Compared to conventional tissue fabrication, which requires longer UV exposure, the viability of the printed cells with simultaneous photopolymerization was significantly higher. Printed neocartilage demonstrated excellent glycosaminoglycan (GAG) and collagen type II production, which was consistent with gene expression. Therefore, this platform is ideal for accurate cell distribution and arrangement for anatomic tissue engineering.

The methods described in this paper show how to convert a commercial inkjet printer into a bioprinter with simultaneous UV polymerization. The printer is capable of constructing 3D tissue structure with cells and biomaterials. The study demonstrated here constructed a 3D neocartilage.

三次元インクジェット印刷技術を用いて、ヒト軟骨組織の作製

Bioprinting, which is based on thermal inkjet printing, is one of the most attractive enabling technologies in the field of tissue engineering and ...

Analysis of Cell Migration within a Three-dimensional Collagen Matrix

The ability to migrate is a hallmark of various cell types and plays a crucial role in several physiological processes, including&#160;embryonic development, wound healing, and immune responses. However, cell migration is also a key mechanism in cancer enabling these cancer cells to detach from the primary tumor to start metastatic spreading. Within the past years various cell migration assays have been developed to analyze the migratory behavior of different cell types. Because the locomotory behavior of cells markedly differs between a two-dimensional (2D) and three-dimensional (3D) environment it can be assumed that the analysis of the migration of cells that are embedded within a 3D environment would yield in more significant cell migration data. The advantage of the described 3D collagen matrix migration assay is that cells are embedded within a physiological 3D network of collagen fibers representing the major component of the extracellular matrix. Due to time-lapse video microscopy real cell migration is measured allowing the determination of several migration parameters as well as their alterations in response to pro-migratory factors or inhibitors. Various cell types could be analyzed using this technique, including lymphocytes/leukocytes, stem cells, and tumor cells. Likewise, also cell clusters or spheroids could be embedded within the collagen matrix concomitant with analysis of the emigration of single cells from the cell cluster/ spheroid into the collagen lattice. We conclude that the 3D collagen matrix migration assay is a versatile method to analyze the migration of cells within a physiological-like 3D environment.

Cell migration is a biological phenomenon that is involved in a plethora of physiological, such as wound healing and immune responses, and pathophysiological processes, like cancer. The 3D-collagen matrix migration assay is a versatile tool to analyze the migratory properties of different cell types within in a 3D physiological-like environment.

三次元コラーゲンマトリックス内の細胞移動の解析

The ability to migrate is a hallmark of various cell types and plays a crucial role in several physiological processes, including&#160;embryonic ...

Generation of a Three-dimensional Full Thickness Skin Equivalent and Automated Wounding

In vitro models are a cost effective and ethical alternative to study cutaneous wound healing processes. Moreover, by using human cells, these models reflect the human wound situation better than animal models. Although two-dimensional models are widely used to investigate processes such as cellular migration and proliferation, models that are more complex are required to gain a deeper knowledge about wound healing. Besides a suitable model system, the generation of precise and reproducible wounds is crucial to ensure comparable results between different test runs. In this study, the generation of a three-dimensional full thickness skin equivalent to study wound healing is shown. The dermal part of the models is comprised of human dermal fibroblast embedded in a rat-tail collagen type I hydrogel. Following the inoculation with human epidermal keratinocytes and consequent culture at the air-liquid interface, a multilayered epidermis is formed on top of the models. To study the wound healing process, we additionally developed an automated wounding device, which generates standardized wounds in a sterile atmosphere.

The goal of this protocol is to build up a three-dimensional full thickness skin equivalent, which resembles natural skin. With a specifically constructed automated wounding device, precise and reproducible wounds can be generated under maintenance of sterility.

三次元の全層皮膚等価物と自動傷つけ処理の発生

In vitro models are a cost effective and ethical alternative to study cutaneous wound healing processes. Moreover, by using human cells, these models ...

Preparation and Analysis of In Vitro Three Dimensional Breast Carcinoma Surrogates

Three dimensional (3D) culture is a more physiologically relevant method to model cell behavior in vitro than two dimensional culture. Carcinomas, including breast carcinomas, are complex 3D tissues composed of cancer epithelial cells and stromal components, including fibroblasts and extracellular matrix (ECM). Yet most in vitro models of breast carcinoma consist only of cancer epithelial cells, omitting the stroma and, therefore, the 3D architecture of a tumor in vivo. Appropriate 3D modeling of carcinoma is important for accurate understanding of tumor biology, behavior, and response to therapy. However, the duration of culture and volume of 3D models is limited by the availability of oxygen and nutrients within the culture. Herein, we demonstrate a method in which breast carcinoma epithelial cells and stromal fibroblasts are incorporated into ECM to generate a 3D breast cancer surrogate that includes stroma and can be cultured as a solid 3D structure or by using a perfusion bioreactor system to deliver oxygen and nutrients. Following setup and an initial growth period, surrogates can be used for preclinical drug testing. Alternatively, the cellular and matrix components of the surrogate can be modified to address a variety of biological questions. After culture, surrogates are fixed and processed to paraffin, in a manner similar to the handling of clinical breast carcinoma specimens, for evaluation of parameters of interest. The evaluation of one such parameter, the density of cells present, is explained, where ImageJ and CellProfiler image analysis software systems are applied to photomicrographs of histologic sections of surrogates to quantify the number of nucleated cells per area. This can be used as an indicator of the change in cell number over time or the change in cell number resulting from varying growth conditions and treatments.

Preparation and Analysis of In Vitro Three Dimensional Breast Carcinoma Surrogates

We demonstrate a method to generate 3D breast cancer surrogates, which can be cultured using a perfusion bioreactor system to deliver oxygen and nutrients. Following growth, surrogates are fixed and processed to paraffin for evaluation of parameters of interest. The evaluation of one such parameter, cell density, is explained.

の調製と分析<em&gt;インビトロ</em&gt;三次元乳癌サロゲート

Three dimensional (3D) culture is a more physiologically relevant method to model cell behavior in vitro than two dimensional culture. Carcinomas, ...

Engineering Three-dimensional Epithelial Tissues Embedded within Extracellular Matrix

The architecture of branched organs such as the lungs, kidneys, and mammary glands arises through the developmental process of branching morphogenesis, which is regulated by a variety of soluble and physical signals in the microenvironment. Described here is a method created to study the process of branching morphogenesis by forming engineered three-dimensional (3D) epithelial tissues of defined shape and size that are completely embedded within an extracellular matrix (ECM). This method enables the formation of arrays of identical tissues and enables the control of a variety of environmental factors, including tissue geometry, spacing, and ECM composition. This method can also be combined with widely used techniques such as traction force microscopy (TFM) to gain more information about the interactions between cells and their surrounding ECM. The protocol can be used to investigate a variety of cell and tissue processes beyond branching morphogenesis, including cancer invasion.

This manuscript describes a soft lithography-based technique to engineer uniform arrays of three-dimensional (3D) epithelial tissues of defined geometry surrounded by extracellular matrix. This method is amenable to a wide variety of cell types and experimental contexts and allows for high-throughput screening of identical replicates.

細胞外マトリックス内に埋め込まれたエンジニアリング三次元上皮組織

The architecture of branched organs such as the lungs, kidneys, and mammary glands arises through the developmental process of branching morphogenesis, ...

Fabrication of Three-dimensional Paper-based Microfluidic Devices for Immunoassays

Paper wicks fluids autonomously due to capillary action. By patterning paper with hydrophobic barriers, the transport of fluids can be controlled and directed within a layer of paper. Moreover, stacking multiple layers of patterned paper creates sophisticated three-dimensional microfluidic networks that can support the development of analytical and bioanalytical assays. Paper-based microfluidic devices are inexpensive, portable, easy to use, and require no external equipment to operate. As a result, they hold great promise as a platform for point-of-care diagnostics. In order to properly evaluate the utility and analytical performance of paper-based devices, suitable methods must be developed to ensure their manufacture is reproducible and at a scale that is appropriate for laboratory settings. In this manuscript, a method to fabricate a general device architecture that can be used for paper-based immunoassays is described. We use a form of additive manufacturing (multi-layer lamination) to prepare devices that comprise multiple layers of patterned paper and patterned adhesive. In addition to demonstrating the proper use of these three-dimensional paper-based microfluidic devices with an immunoassay for human chorionic gonadotropin (hCG), errors in the manufacturing process that may result in device failures are discussed. We expect this approach to manufacturing paper-based devices will find broad utility in the development of analytical applications designed specifically for limited-resource settings.

We detail a method to fabricate three-dimensional paper-based microfluidic devices for use in the development of immunoassays. Our approach to device assembly is a type of multilayer, additive manufacturing. We demonstrate a sandwich immunoassay to provide representative results for these types of paper-based devices.

イムノアッセイのための3次元紙ベースのマイクロ流体デバイスの作製

Paper wicks fluids autonomously due to capillary action. By patterning paper with hydrophobic barriers, the transport of fluids can be controlled and ...

Three-dimensional Printing of Thermoplastic Materials to Create Automated Syringe Pumps with Feedback Control for Microfluidic Applications

Microfluidics has become a critical tool in research across the biological, chemical, and physical sciences. One important component of microfluidic experimentation is a stable fluid handling system capable of accurately providing an inlet flow rate or inlet pressure. Here, we have developed a syringe pump system capable of controlling and regulating the inlet fluid pressure delivered to a microfluidic device. This system was designed using low-cost materials and additive manufacturing principles, leveraging three-dimensional (3D) printing of thermoplastic materials and off-the-shelf components whenever possible. This system is composed of three main components: a syringe pump, a pressure transducer, and a programmable microcontroller. Within this paper, we detail a set of protocols for fabricating, assembling, and programming this syringe pump system. Furthermore, we have included representative results that demonstrate high-fidelity, feedback control of inlet pressure using this system. We expect this protocol will allow researchers to fabricate low-cost syringe pump systems, lowering the entry barrier for the use of microfluidics in biomedical, chemical, and materials research.

Here we present a protocol to construct a pressure-controlled syringe pump to be used in microfluidic applications. This syringe pump is made from an&#160;additively manufactured body, off-the-shelf hardware, and open-source electronics. The resulting system is low-cost, straightforward to build, and delivers well-regulated fluid flow to enable rapid microfluidic research.

印刷作成する熱可塑性材料の三次元マイクロ流体用フィードバック制御を用いたシリンジ ポンプを自動

Microfluidics has become a critical tool in research across the biological, chemical, and physical sciences. One important component of microfluidic ...