我々は技術支援のためにくん呂とロナルドクルツに感謝します。この作品は、ONRのMURI賞号によってサポートされていました：N00014 - 10 - 1から0072まで、NSFの助成＃SBE - 0542013学習センター、学習センターのNSFの科学、およびNIHの助成第2位R01 NS036449 - 11の空間ダイナミクスに。

1。スキャナ室の外<ol><li>サポートされて自由端と切り離さ長いハンドルの外側の端でローリングテーブルに置きます。 </li><li>ロボットがオフになっていることを確認してください。 </li><li>テーブルのソケットにロボットを置き、2本のネジでロボットを介してアルミニウムの安全性のプレートを固定します。 </li><li>アルミアダプター付きロボットハンドルにエンドエフェクタを取り付け、それが自由に動くことを確認してください。 </li><li>アルミロボットにシールドと10&#39;のパラレルケーブルを接続し、シールドが損なわれていないことを確認してください。必要に応じて余分な箔を追加。 </li><li>背面の溝に平行ケーブルと電源ケーブルを配置するように注意してロボットを介しアルミシールドボックスを置きます。 </li><li>シールドボックスに慎重にねじ込みます。 </li><li>シールドボックスのケーブル溝にアルミホイルを梱包し、箔がパラレルケーブルにシールドに接触することを確認してください。 </li></ol> 2。二人、AとBとスキャナの部屋に移動する<ol><li>あらゆる非鉄もの、例えば、携帯電話、鍵、硬貨等を含むすべての金属物を除去することにより、高磁場環境を入力するための準備.. </li><li>人とロボットまで、最初に、ロボットのテーブルやボックスの端を安定させる人物Bの自由端をロール保持自由端は、ちょうど部屋のドアに入る。 </li><li>ユーザーBのクリップは、セキュリティのロープは、もう一方の端がしっかりと壁のアンカーに接続されているシールドケースと小切手の裏面にアンカー穴につなが。 </li><li>一緒に働いて、部屋にテーブルをロールバックし、fMRIのテーブルの足にマジックテープのストラップで、それを添付。テーブルのロボットの終わりには、可能な限り、スキャナから滞在す​​る必要があります。 </li><li>ロボットでのカスタムコントロールルームに至るまでのパスでフィルタ、およびプラグにロボットのパラレルケーブルを接続します。パラレルケーブルのシールド箔は、フィルターとの接触を確認する必要があります。 </li><li>長いハンドルの外側の部分（エンドエフェクタ）を添付し、それが正常に機能的MRIのボアを入力することを確認してください。 </li></ol> 3。制御室で<ol><li>制御用コンピュータを起動して、ロボットに6&#39;パラレルケーブルを取り付けます。パススルーのインテリアの一部になる可能性のある追加のフィルタを削除してください。 </li><li>ロボットのモータがオフになっていることを確認する、ファントムキャリブレーションルーチンを起動して、ロボットのキャリブレーションルーチンからモータの位置の読み出しが安定していることを確認してください。 </li><li>パラレルケーブル接続をチェックし、キャリブレーションルーチンがロボットを見ることができないまたはモーターの読み出しに大きな変動がある場合にのみ、大規模なカスタムフィルタが接続されていることをダブルクリック。 </li><li>シールドボックスの背面のポートを開くことで、ロボットの電源を入れ、棒でスイッチを押してください。 </li><li>略円錐導波路の端を中心とするエンドエフェクタを持つキャリブレーションルーチンを再起動します。キャリブレーションをステップ実行し、ボックスのキャリブレーションボックスが適切なハプティックインタラクションを持っていることを確認してください。 </li><li>制御用コンピュータで機能的MRI（BNCコネクタ）からLabjack ADCへのTTL出力を取り付けます。 </li></ol> 4。対象と<ol><li>標準的なfMRIのプロトコルと高磁場環境のための主題を準備します。 </li><li>実験、例えば、視覚的な表示システムのための余分な機器をセットアップします。我々は、視覚的なディスプレイのステレオ表示、仮想環境を提示するときに特に便利です機能を提供するNordicNeuroラボ、（株）ビジョンシステムを使用してください。 </li><li>被写体がテーブルの上に横になると、ヘッドのコイルが位置された後、手のネジを緩め、被験者が快適に移動できるようになるまでテーブルの上をスライドさせて、ロボットの距離を調整する。 </li><li> fMRIの表は、ウォブリングからキャスターを防ぐために、ボアのうちに動いている間にロボットの端に保持することによって手動でテーブルをご案内。エンドエフェクタは、穴に入ると外でキャッチしていないことを確認してください。 </li><li>実験を実行します。 </li></ol> 5。二人とBとのセットアップを打破する<ol><li>患者が終了した後、長いハンドルの外側の端を削除し、ベルクロストラップを元に戻すことによってfMRIのテーブルからロボットのテーブルを切り離します。 </li><li>ロボットから、シールドパラレルケーブルを元に戻すと電源ケーブルを抜いてください。 </li><li>人とロボットのテーブルとロボットの終了を導く人物Bの自由端を持って、ドアにテーブルを移動する。玄関で、テザーを取り消し、そしてホールにロボットのテーブルをロールアウト。 </li><li>すべてのシールドボックスからネジと安全性のプレートから2本のネジを元に戻すとロボットを取り外します。 </li></ol> 6。代表的な結果： 理想的には、触覚ロボットとfMRIはお互いに影響を与えないはずです。ロボットは、fMRIによる影響を受けている場合我々はオンラインで伝えることができます。ロボットのパラレルケーブルが適切に遮蔽され、フィルタリングされていない場合は一般的に、、その後、モーターの読み出しは、発振される後半急速に。これは、鉄のコアが適切にロボットの近くにパラレルケーブル上に配置されていること、二重ケーブルのシールドアルミをチェックすることで固定し、ロボットにのみフィルタすることは、パススルーのスキャナの部屋側のカスタムフィルタですできます。 fMRIの中のエラーを検出すると、データが減少して分析された後に実際には可能ですが、解剖学的スキャンは、初期の研究で採取し、相関ノイズ（例えばスパイクノイズ）7の示すジッパーの効果や他のアーティファクトをチェックする必要があります。頻繁に、このようなノイズは、金属の接点に金属から来ていると、特にロボットのテーブルのすべてのネジ、テーブルの側の手の調整ネジを締めてクリーンアップすることができます。我々のテストから雑音比にベースラインのfMRIの信号（SNR）〜380と完全にはまだ合理的〜330まで下がる部屋にシールドロボットとなります。シールドはロボット上の場所にない場合は、〜250、およびノイズの影響をさらにSNRの低下は非常に重要になります。 4に示すように、ハンドルの中心にある自由のジョイントの3度はロボットから離れてシフトすることを除き、ロボット/ハンド相互作用の動力学にほとんど影響を与えません。ハンドルの中央のジョイントが支点のように動作し、向きは2（左右、上下）が、サードではない（将来のバック）で明らかに動きが逆になります。ファントムと手が途中でその支点とハンドルのようなレバーの反対側の端にあるので、利益は3つのユークリッド方向のそれぞれにソフトウェアで適用されます。スイベルジョイントと正のゲインで制御される二つの方向に負の利益スライダージョイントの方向です。ハンドルとスイベルの正味の効果は、ファントムのロボットの自由度、車でわずか9&#39;の完全な3度を再現。 <img src="/files/ftp_upload/3364/3364fig1.jpg" alt="図1"/> 図1 fMRIの環境で使用するための触覚ロボットをマウントするために使用される装置。トップへ前の筐体（左、上）とハンドルの中間点（右、上）での共同ジンバル/スライダーのケースに実装されている触覚ロボットを示しています。左下には、ハンドルを操作するスキャナの主題を示しています。底には、右のシールドと終了エフェクタの漫画です。 <img src="/files/ftp_upload/3364/3364fig2.jpg" alt="図2"/> 動きと完全にシールドされたロボットのためのBIRN試験の2の結果図 。十字架と3つの画像は、球状の頭部モデルのセクションを示し、そして右下には、3次元のビューを示しています。小さな丸は、静的な頭部モデルにおける気泡であり、常に存在している。大きなストライプやジッパーの欠如は、ロボットからのノイズが相関していることを示します。

<ol>
	<li>Ogawa, S., Lee, T. M., Kay, A. R., Tank, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brain+magnetic+resonance+imaging+with+contrast+dependent+on+blood+oxygenation.">Brain magnetic resonance imaging with contrast dependent on blood oxygenation.</a> Proc. Natl. Acad. Sci. U.S.A. 87, 9868-9872 (1990).</li><li>Heeger, D. J., Ress, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=What+does+fMRI+tell+us+about+neuronal+activity.">What does fMRI tell us about neuronal activity.</a> Nat. Rev. Neurosci. 3, 142-151 (2002).</li><li>deCharms, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Applications+of+real-time+fMRI.">Applications of real-time fMRI.</a> Nat. Rev. Neurosci. 9, 720-729 (2008).</li><li>Hribar, A., Koritnik, B., Munih, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phantom+haptic+device+upgrade+for+use+in+fMRI.">Phantom haptic device upgrade for use in fMRI.</a> Medical and Biological Engineering and Computing. 47, 677-684 (2009).</li><li>Trommershauser, J., Gepshtein, S., Maloney, L. T., Landy, M. S., Banks, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimal+compensation+for+changes+in+task-relevant+movement+variability.">Optimal compensation for changes in task-relevant movement variability.</a> J. Neurosci. 25, 7169-7178 (2005).</li><li>Konczak, J., Li, K. Y., Tuite, P. J., Poizner, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Haptic+perception+of+object+curvature+in+Parkinson's+disease.">Haptic perception of object curvature in Parkinson's disease.</a> PLoS ONE. 3, e2625-e2625 (2008).</li><li>Lipton, M. L., Lipton, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Artifacts:+When+things+go+wrong,+it's+not+necessarily+all+bad.">Artifacts: When things go wrong, it's not necessarily all bad.</a> Totally Accessible MRI. , 139-153 (2008).</li><li>Rajh, M., Glodez, S., Flasker, J., Gotlih, K., Kostanjevec, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design+and+analysis+of+an+fMRI+compatible+haptic+robot.">Design and analysis of an fMRI compatible haptic robot.</a> Robotics and Computer-Integrated Manufacturing. 27, 267-275 (2011).</li><li>Burdet, E., Gassert, R., Gowrishankar, G., Chapuis, D., Bleuler, H., Ang, M., Khatib, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=fMRI+compatible+haptic+interfaces+to+investigate+human+motor+control.">fMRI compatible haptic interfaces to investigate human motor control.</a> Experimental Robotics IX, of Springer Tracts in Advanced Robotics. 21, 25-34 (2006).</li><li>Nakamura, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+networks+in+motor+sequence+learning:+abnormal+topographies+in+Parkinson's+disease.">Functional networks in motor sequence learning: abnormal topographies in Parkinson's disease.</a> Hum Brain Mapp. 12, 42-60 (2001).</li></ol>

利害の衝突は宣言されません。

fMRIの互換性のあるロボットは、モータ制御の神経科学の実験のための新たな可能性を開きます。セットアップで最も重要なステップは、我々は二つのステップで行う機能的MRI、内のアーティファクトを防ぐために、ロボットのシールドです。最初に、ロボット自体が長く、軽量、自由関節の3度と、その途中でサポートされているハンドルを持つボアから離れて約9&#39;です。第二に、ロボッ?...

<table border="1"><tbody><tr><td>ファントムプレミアム1.5/6dof、高い力のモデル</td><td> Sensable </td><td> <a href="www.sensable.com">www.sensable.com</a> </td></tr></tbody></table>

adaptation of a haptic robot in a 3t fmri

機能的磁気共鳴画像（fMRI）は非電離放射線、解剖学的および機能的なデータのミリ空間精度2、ほぼリアルタイムの分析3を含む利点とBOLD信号1を介して、優れた脳機能イメージングを提供します。ハプティックロボットは、合理的に限定されたスペース内でカーソルの位置と力の精密計測と制御を提供しています。ここでは、触覚/などに到達するか、または把握のような触覚環境の相互作用によるモータ制御に関わる精密実験を可能にするためにこれら二つの技術を組み合わせる。基本的な考え方は図1（被験者がロボットを使用できるようにロボットは4〜中央でサポートされている8フィートのエンドエフェクタを添付することですが、それを遮蔽し、fMRIのマシンからの磁界の最も極端な部分から、それを締め出す）。 ファントムプレミアム3.0、6自由度、高フォースロボットは（SensAble Technologies社）バーチャルリアリティの実験5,6で、フォースフィードバックを提供するための優れた選択肢ですが、それは本質的に非MR安全です、敏感に大きなノイズを紹介します機能的MRI装置、およびその電気モーターが強く磁場を変化させてfMRIによる影響を受ける可能性があります。我々は、ロボットが安全に機能的MRI環境に導入することを可能にするとの大きく変化する磁場によって電気的ノイズの多いモーターによるfMRI信号の劣化と電気モーターの性能の劣化の両方を制限するテーブルとシールドシステムを構築しています機能的MRI。シールドと、信号対雑音比（SNR：平均シグナル/ノイズの標準偏差）fMRIのは、〜380のベースラインから〜330、およびシールドなし〜250になります。残りのノイズが無相関であるために表示され、試験球の機能的MRI（図2）にアイテムを追加していません。ロボットの機能的MRIの有意な効果がないので、長く、硬いハンドルが磁場の中で最も大きく変化する部​​分の範囲外のロボットの配置を可能に。ロボットの運動学上のハンドルの効果は、それが軽量であるため、最小限のです（〜2.6ポンド）が非常に堅い3 / 4&quot;グラファイトと真ん中に3自由度関節の上にバランスよく。と最終結果は、fMRIの互換性、触覚システムです。作業スペースの約1立方フィート、そして、バーチャルリアリティと組み合わせると、それは様々な力の場での適応学習、手足や触覚知覚の自然な遠大な、受動的な変位を含むfMRIの環境で実行される実験の新しいセットが可能、またはテクスチャ識別5,6。

Watch this Scientific Journal Video about Adaptation of a Haptic Robot in a 3T fMRI at JoVE.com

Adaptation of a Haptic Robot in a 3T fMRI

3TのfMRIの中の触覚ロボットの適応と使用が記載されている。

3TのfMRIの中の触覚ロボットの適応

Functional magnetic resonance imaging (fMRI) provides excellent functional brain imaging via the BOLD signal 1 with advantages including non-ionizing ...

adaptation-of-a-haptic-robot-in-a-3t-fmri

Research

JoVE Journal

Bioengineering

9.6K ビュー.  University of California. 3TのfMRIの中の触覚ロボットの適応と使用が記載されている。

ビデオ: Adaptation of a Haptic Robot in a 3T fMRI

Fabrication of Electrochemical-DNA Biosensors for the Reagentless Detection of Nucleic Acids, Proteins and Small Molecules

As medicine is currently practiced, doctors send specimens to a central laboratory for testing and thus must wait hours or days to 
receive the results. Many patients would be better served by rapid, bedside tests. To this end our laboratory and others have developed a versatile, reagentless 
biosensor platform that supports the quantitative, reagentless, electrochemical detection of nucleic acids (DNA, RNA), proteins (including antibodies) and small 
molecules analytes directly in unprocessed clinical and environmental samples. In this video, we demonstrate the preparation and use of several biosensors in this 
"E-DNA" class. In particular, we fabricate and demonstrate sensors for the detection of a target DNA sequence in a polymerase chain reaction mixture, an HIV-specific antibody and the drug cocaine. The preparation procedure requires only three hours of hands-on effort followed by an overnight incubation, and their use requires only minutes.

"E-DNA" sensors, reagentless, electrochemical biosensors that perform well even when challenged directly in blood and other complex matrices, have been adapted to the detection of a wide range of nucleic acid, protein and small molecule analytes. Here we present a general procedure for the fabrication and use of such sensors.

核酸、タンパク質と低分子のReagentless検出用電気化学DNAバイオセンサーの作製

As medicine is currently practiced, doctors send specimens to a central laboratory for testing and thus must wait hours or days to 
receive the results. ...

生物工学

Haptic/Graphic Rehabilitation: Integrating a Robot into a Virtual Environment Library and Applying it to Stroke Therapy

Recent research that tests interactive devices for prolonged therapy practice has revealed new prospects for robotics combined with graphical and other forms of biofeedback. Previous human-robot interactive systems have required different software commands to be implemented for each robot leading to unnecessary developmental overhead time each time a new system becomes available. For example, when a haptic/graphic virtual reality environment has been coded for one specific robot to provide haptic feedback, that specific robot would not be able to be traded for another robot without recoding the program. However, recent efforts in the open source community have proposed a wrapper class approach that can elicit nearly identical responses regardless of the robot used. The result can lead researchers across the globe to perform similar experiments using shared code. Therefore modular "switching out"of one robot for another would not affect development time. In this paper, we outline the successful creation and implementation of a wrapper class for one robot into the open-source H3DAPI, which integrates the software commands most commonly used by all robots.

Recently, a vast amount of prospects have come available for human-robot interactive systems. In this paper we outline the integration of a new robotic device with open source software that can rapidly make possible a library of interactive functionality. We then outline a clinical application for a neurorehabilitation application.

触覚/グラフィックリハビリテーション：仮想環境のライブラリにロボットを統合し、脳卒中治療に適用すること

Recent research that tests interactive devices for prolonged therapy practice has revealed new prospects for robotics combined with graphical and other ...

Creating Two-Dimensional Patterned Substrates for Protein and Cell Confinement

Microcontact printing provides a rapid, highly reproducible method for the creation of well-defined patterned substrates.1 While microcontact printing can be employed to directly print a large number of molecules, including proteins,2 DNA,3 and silanes,4 the formation of self-assembled monolayers (SAMs) from long chain alkane thiols on gold provides a simple way to confine proteins and cells to specific patterns containing adhesive and resistant regions. This confinement can be used to control cell morphology and is useful for examining a variety of questions in protein and cell biology. Here, we describe a general method for the creation of well-defined protein patterns for cellular studies.5 This process involves three steps: the production of a patterned master using photolithography, the creation of a PDMS stamp, and microcontact printing of a gold-coated substrate. Once patterned, these cell culture substrates are capable of confining proteins and/or cells (primary cells or cell lines) to the pattern.
The use of self-assembled monolayer chemistry allows for precise control over the patterned protein/cell adhesive regions and non-adhesive regions; this cannot be achieved using direct protein stamping. Hexadecanethiol, the long chain alkane thiol used in the microcontact printing step, produces a hydrophobic surface that readily adsorbs protein from solution. The glycol-terminated thiol, used for backfilling the non-printed regions of the substrate, creates a monolayer that is resistant to protein adsorption and therefore cell growth.6 These thiol monomers produce highly structured monolayers that precisely define regions of the substrate that can support protein adsorption and cell growth. As a result, these substrates are useful for a wide variety of applications from the study of intercellular behavior7 to the creation of microelectronics.8
While other types of monolayer chemistry have been used for cell culture studies, including work from our group using trichlorosilanes to create patterns directly on glass substrates,9 patterned monolayers formed from alkane thiols on gold are straight-forward to prepare. Moreover, the monomers used for monolayer preparation are commercially available, stable, and do not require storage or handling under inert atmosphere. Patterned substrates prepared from alkane thiols can also be recycled and reused several times, maintaining cell confinement.10

Self-assembled monolayers (SAMs) formed from long chain alkane thiols on gold provide well-defined substrates for the formation of protein patterns and cell confinement. Microcontact printing of hexadecanethiol using a polydimethylsiloxane (PDMS) stamp followed by backfilling with a glycol-terminated alkane thiol monomer produces a pattern where protein and cells adsorb only to the stamped hexadecanethiol region.

タンパク質、細胞閉じ込めのための二次元加工基板を作成する

Microcontact printing provides a rapid, highly reproducible method for the creation of well-defined patterned substrates.1  While microcontact printing ...

Cellular Lipid Extraction for Targeted Stable Isotope Dilution Liquid Chromatography-Mass Spectrometry Analysis

The metabolism of fatty acids, such as arachidonic acid (AA) and linoleic acid (LA), results in the formation of oxidized bioactive lipids, including numerous stereoisomers1,2. These metabolites can be formed from free or esterified fatty acids. Many of these oxidized metabolites have biological activity and have been implicated in various diseases including cardiovascular and neurodegenerative diseases, asthma, and cancer3-7. Oxidized bioactive lipids can be formed enzymatically or by reactive oxygen species (ROS). Enzymes that metabolize fatty acids include cyclooxygenase (COX), lipoxygenase (LO), and cytochromes P450 (CYPs)1,8. Enzymatic metabolism results in enantioselective formation whereas ROS oxidation results in the racemic formation of products.
While this protocol focuses primarily on the analysis of AA- and some LA-derived bioactive metabolites; it could be easily applied to metabolites of other fatty acids. Bioactive lipids are extracted from cell lysate or media using liquid-liquid (l-l) extraction. At the beginning of the l-l extraction process, stable isotope internal standards are added to account for errors during sample preparation. Stable isotope dilution (SID) also accounts for any differences, such as ion suppression, that metabolites may experience during the mass spectrometry (MS) analysis9. After the extraction, derivatization with an electron capture (EC) reagent, pentafluorylbenzyl bromide (PFB) is employed to increase detection sensitivity10,11. Multiple reaction monitoring (MRM) is used to increase the selectivity of the MS analysis. Before MS analysis, lipids are separated using chiral normal phase high performance liquid chromatography (HPLC). The HPLC conditions are optimized to separate the enantiomers and various stereoisomers of the monitored lipids12. This specific LC-MS method monitors prostaglandins (PGs), isoprostanes (isoPs), hydroxyeicosatetraenoic acids (HETEs), hydroxyoctadecadienoic acids (HODEs), oxoeicosatetraenoic acids (oxoETEs) and oxooctadecadienoic acids (oxoODEs); however, the HPLC and MS parameters can be optimized to include any fatty acid metabolites13.
Most of the currently available bioanalytical methods do not take into account the separate quantification of enantiomers. This is extremely important when trying to deduce whether or not the metabolites were formed enzymatically or by ROS. Additionally, the ratios of the enantiomers may provide evidence for a specific enzymatic pathway of formation. The use of SID allows for accurate quantification of metabolites and accounts for any sample loss during preparation as well as the differences experienced during ionization. Using the PFB electron capture reagent increases the sensitivity of detection by two orders of magnitude over conventional APCI methods. Overall, this method, SID-LC-EC-atmospheric pressure chemical ionization APCI-MRM/MS, is one of the most sensitive, selective, and accurate methods of quantification for bioactive lipids.

This protocol will demonstrate the extraction and analysis of free and esterified bioactive fatty acids from cells. Fatty acids are accurately quantified using stable isotope dilution, chiral liquid chromatography, electron capture atmospheric chemical ionization multiple reaction monitoring mass spectrometry (SID-LC-ECAPCI-MRM/MS).

ターゲット安定同位体希釈液体クロマトグラフィー - 質量分析のための細胞の脂質の抽出

The metabolism of fatty acids, such as arachidonic acid (AA) and linoleic acid (LA), results in the formation of oxidized bioactive lipids, including ...

Designing a Bio-responsive Robot from DNA Origami

Nucleic acids are astonishingly versatile. In addition to their natural role as storage medium for biological information1, they can be utilized in parallel computing2,3 , recognize and bind molecular or cellular targets4,5 , catalyze chemical reactions6,7 , and generate calculated responses in a biological system8,9. Importantly, nucleic acids can be programmed to self-assemble into 2D and 3D structures10-12, enabling the integration of all these remarkable features in a single robot linking the sensing of biological cues to a preset response in order to exert a desired effect.
Creating shapes from nucleic acids was first proposed by Seeman13, and several variations on this theme have since been realized using various techniques11,12,14,15 . However, the most significant is perhaps the one proposed by Rothemund, termed scaffolded DNA origami16. In this technique, the folding of a long (&gt;7,000 bases) single-stranded DNA 'scaffold' is directed to a desired shape by hundreds of short complementary strands termed 'staples'. Folding is carried out by temperature annealing ramp. This technique was successfully demonstrated in the creation of a diverse array of 2D shapes with remarkable precision and robustness. DNA origami was later extended to 3D as well17,18 .
 
The current paper will focus on the caDNAno 2.0 software19 developed by Douglas and colleagues. caDNAno is a robust, user-friendly CAD tool enabling the design of 2D and 3D DNA origami shapes with versatile features. The design process relies on a systematic and accurate abstraction scheme for DNA structures, making it relatively straightforward and efficient.
In this paper we demonstrate the design of a DNA origami nanorobot that has been recently described20. This robot is 'robotic' in the sense that it links sensing to actuation, in order to perform a task. We explain how various sensing schemes can be integrated into the structure, and how this can be relayed to a desired effect. Finally we use Cando21 to simulate the mechanical properties of the designed shape. The concept we discuss can be adapted to multiple tasks and settings.

DNA origami is a powerful method for fabricating precise nanoscale objects by programming the self-assembly of DNA molecules. Here, we describe how DNA origami can be utilized to design a robotic robot capable of sensing biological cues and responding by shape shifting, subsequently relayed to a desired effect.

DNA折り紙からバイオ応答ロボットの設計

Nucleic acids are astonishingly versatile. In addition to their natural role as storage medium for biological information1, they can be utilized in ...

Identification of Metal Oxide Nanoparticles in Histological Samples by Enhanced Darkfield Microscopy and Hyperspectral Mapping

Nanomaterials are increasingly prevalent throughout industry, manufacturing, and biomedical research. The need for tools and techniques that aid in the identification, localization, and characterization of nanoscale materials in biological samples is on the rise. Currently available methods, such as electron microscopy, tend to be resource-intensive, making their use prohibitive for much of the research community. Enhanced darkfield microscopy complemented with a hyperspectral imaging system may provide a solution to this bottleneck by enabling rapid and less expensive characterization of nanoparticles in histological samples. This method allows for high-contrast nanoscale imaging as well as nanomaterial identification. For this technique, histological tissue samples are prepared as they would be for light-based microscopy. First, positive control samples are analyzed to generate the reference spectra that will enable the detection of a material of interest in the sample. Negative controls without the material of interest are also analyzed in order to improve specificity (reduce false positives). Samples can then be imaged and analyzed using methods and software for hyperspectral microscopy or matched against these reference spectra in order to provide maps of the location of materials of interest in a sample. The technique is particularly well-suited for materials with highly unique reflectance spectra, such as noble metals, but is also applicable to other materials, such as semi-metallic oxides. This technique provides information that is difficult to acquire from histological samples without the use of electron microscopy techniques, which may provide higher sensitivity and resolution, but are vastly more resource-intensive and time-consuming than light microscopy.

Enhanced darkfield microscopy and hyperspectral imaging with spectral mapping enable screening, localization, and identification of nanoscale materials in histological samples with improved speed and accuracy over traditional methods. The goal of this paper is to provide methods for darkfield imaging and hyperspectral mapping of metal oxide nanoparticles in histological samples.

強化された暗視野顕微鏡とハイパースペクトルマッピングにより組織学的試料中の金属酸化物ナノ粒子の同定

Nanomaterials are increasingly prevalent throughout industry, manufacturing, and biomedical research. The need for tools and techniques that aid in the ...

Medical-grade Sterilizable Target for Fluid-immersed Fetoscope Optical Distortion Calibration

We have developed a calibration target for use with fluid-immersed endoscopes within the context of the GIFT-Surg (Guided Instrumentation for Fetal Therapy and Surgery) project. One of the aims of this project is to engineer novel, real-time image processing methods for intra-operative use in the treatment of congenital birth defects, such as spina bifida and the twin-to-twin transfusion syndrome. The developed target allows for the sterility-preserving optical distortion calibration of endoscopes within a few minutes. Good optical distortion calibration and compensation are important for mitigating undesirable effects like radial distortions, which not only hamper accurate imaging using existing endoscopic technology during fetal surgery, but also make acquired images less suitable for potentially very useful image computing applications, like real-time mosaicing. In this paper proposes a novel fabrication method to create an affordable, sterilizable calibration target suitable for use in a clinical setup. This method involves etching a calibration pattern by laser cutting a sandblasted stainless steel sheet. This target was validated using the camera calibration module provided by OpenCV, a state-of-the-art software library popular in the computer vision community.

This article describes the design and development of a sterilizable custom camera optical distortion calibration target for the peri-operative, fluid-immersed calibration of endoscopes during endoscopic interventions.

流体浸漬胎児鏡光学歪み校正のための医療用滅菌可能なターゲット

We have developed a calibration target for use with fluid-immersed endoscopes within the context of the GIFT-Surg (Guided Instrumentation for Fetal ...

Optimizing the Use of a Liquid Handling Robot to Conduct a High Throughput Forward Chemical Genetics Screen of Arabidopsis thaliana

Chemical genetics is increasingly being employed to decode traits in plants that may be recalcitrant to traditional genetics due to gene redundancy or lethality. However, the probability of a synthetic small molecule being bioactive is low; therefore, thousands of molecules must be tested in order to find those of interest. Liquid handling robotics systems are designed to handle large numbers of samples, increasing the speed with which a chemical library can be screened in addition to minimizing/standardizing error. To achieve a high-throughput forward chemical genetics screen of a library of 50,000 small molecules on Arabidopsis thaliana (Arabidopsis), protocols using a bench-top multichannel liquid handling robot were developed that require minimal technician involvement. With these protocols, 3,271 small molecules were discovered that caused visible phenotypic alterations. 1,563 compounds induced short roots, 1,148 compounds altered coloration, 383 compounds caused root hair and other, non-categorized, alterations, and 177 compounds inhibited germination.

Optimizing the Use of a Liquid Handling Robot to Conduct a High Throughput Forward Chemical Genetics Screen of Arabidopsis thaliana

A high throughput screen of synthetic small molecules was conducted on the model plant species, Arabidopsis thaliana. This protocol, developed for a liquid handling robot, increases the speed of forward chemical genetics screens, accelerating the discovery of novel small molecules affecting plant physiology.

高スループット転送化学遺伝学の画面シロイヌナズナを行う液体処理ロボットの使用を最適化します。

Chemical genetics is increasingly being employed to decode traits in plants that may be recalcitrant to traditional genetics due to gene redundancy or ...

Decellularization of Whole Human Heart Inside a Pressurized Pouch in an Inverted Orientation

The ultimate solution for patients with end-stage heart failure is organ transplant. But donor hearts are limited, immunosuppression is required, and ultimately rejection can occur. Creating a functional, autologous bio-artificial heart could solve these challenges. Biofabrication of a heart comprised of scaffold and cells is one option. A natural scaffold with tissue-specific composition as well as micro- and macro-architecture can be obtained by decellularizing hearts from humans or large animals such as pigs. Decellularization involves washing out cellular debris while preserving 3D extracellular matrix and vasculature and allowing &#34;cellularization&#34; at a later timepoint. Capitalizing on our novel finding that perfusion decellularization of complex organs is possible, we developed a more &#34;physiological&#34; method to decellularize non-transplantable human hearts by placing them inside a pressurized pouch, in an inverted orientation, under controlled pressure. The purpose of using a pressurized pouch is to create pressure gradients across the aortic valve to keep it closed and improve myocardial perfusion. Simultaneous assessment of flow dynamics and cellular debris removal during decellularization allowed us to monitor both fluid inflow and debris outflow, thereby generating a scaffold that can be used either for simple cardiac repair (e.g. as a patch or valve scaffold) or as a whole-organ scaffold.

This method enables decellularization of a complex solid organ using a simple protocol based on osmotic shock and perfusion of ionic detergent with minimal organ matrix disruption. It comprises a novel decellularization technique for human hearts inside a pressurized pouch with real-time monitoring of flow dynamics and cellular debris outflow.

逆方向に加圧ポーチ内全体の人間の心の decellularization

The ultimate solution for patients with end-stage heart failure is organ transplant. But donor hearts are limited, immunosuppression is required, and ...

A Tripeptide-Stabilized Nanoemulsion of Oleic Acid

We describe a method to produce a nanoemulsion composed of an oleic acids-Pt(II) core and a lysine-tyrosine-phenylalanine (KYF) coating (KYF-Pt-NE). The KYF-Pt-NE encapsulates Pt(II) at 10 wt. %, has a diameter of 107 &#177; 27 nm and a negative surface charge. The KYF-Pt-NE is stable in water and in serum, and is biologically active. The conjugation of a fluorophore to KYF allows the synthesis of a fluorescent nanoemulsion that is suitable for biological imaging. The synthesis of the nanoemulsion is performed in an aqueous environment, and the KYF-Pt-NE forms via self-assembly of a short KYF peptide and an oleic acids-platinum(II) conjugate. The self-assembly process depends on the temperature of the solution, the molar ratio of the substrates, and the flow rate of the substrate addition. Crucial steps include maintaining the optimal stirring rate during the synthesis, permitting sufficient time for self-assembly, and pre-concentrating the nanoemulsion gradually in a centrifugal concentrator.

This protocol describes an efficient method to synthesize a nanoemulsion of an oleic acids-platinum(II) conjugate stabilized with a lysine-tyrosine-phenylalanine (KYF) tripeptide. The nanoemulsion forms under mild synthetic conditions via self-assembly of the KYF and the conjugate.

オレイン酸のトリペプチド安定ナノエマルション

We describe a method to produce a nanoemulsion composed of an oleic acids-Pt(II) core and a lysine-tyrosine-phenylalanine (KYF) coating (KYF-Pt-NE). The ...