Vorremmo ringraziare Lu Kun e Ronald Kurz per l&#39;assistenza tecnica. Questo lavoro è stato sostenuto da n ° Premio MURI ONR: N00014-10-1-0072, NSF concedere # SBE-0542013 alla dinamica temporale del Learning Center, un Science Learning Center di NSF, NIH e concedere # 2 NS036449 R01-11.

1. Fuori dalla stanza dello scanner <ol><li> Posizionare il tavolo a rotazione con l&#39;estremità libera sostenuto e alla fine esterna del lungo manico staccato. </li><li> Controllare che il robot è spento. </li><li> Posizionare il robot nella presa tavolo e fissare la piastra di sicurezza in alluminio sopra il robot con 2 viti. </li><li> Fissare il effecter fine alla maniglia del robot con l&#39;adattatore in alluminio e verificare che si muove liberamente. </li><li> Collegare il 10 &#39;cavo parallelo con schermatura in alluminio al robot e controllare che la schermatura è intatto. Aggiungi foglio supplementare, se necessario. </li><li> Posizionare la scatola di alluminio schermante sopra il robot avendo cura di posizionare il parallelo e cavi di alimentazione nella scanalatura sul retro. </li><li> Attentamente vite sulla scatola schermatura. </li><li> Confezione foglio di alluminio nella scanalatura del cavo sulla scatola schermatura e assicurarsi che il foglio entra in contatto con la schermatura del cavo parallelo. </li></ol> 2. Lo spostamento nella stanza dello scanner con due persone, A e B <ol><li> Preparatevi ad entrare in un ambiente di alto campo magnetico, eliminando qualsiasi e tutti gli oggetti metallici non ferrosi, compresi quelli, ad esempio, telefoni cellulari, chiavi, monete, ecc .. </li><li> Con la persona A fine tenendo l&#39;estremità libera del tavolo robot e la persona B stabilizzare la fine scatola, rotolare libero prima fino a quando il robot entra solo la porta della stanza. </li><li> B clip persona alla fune di sicurezza legato ad un foro di ancoraggio sul retro della cassa schermatura e controlla che l&#39;altra estremità è saldamente attaccato ad una parete di ancoraggio. </li><li> Lavorando insieme, ruotare la tavola nella stanza e fissarla con le cinghie in velcro ai piedi del tavolo fMRI. La fine robot del tavolo deve rimanere il più dallo scanner come possibile. </li><li> Collegare il cavo parallelo del robot per il filtro personalizzato sul passaggio alla sala di controllo, e collegare il robot. La pellicola schermatura del cavo parallelo deve prendere contatto con il filtro. </li><li> Fissare la parte esterna del lungo manico (effecter fine) e verificare che entrerà nel foro della fMRI pulito. </li></ol> 3. Nella sala di controllo <ol><li> Avviare il computer di controllo e collegare il cavo parallelo 6 &#39;al robot. Accertarsi di rimuovere eventuali filtri aggiuntivi che possono essere sulla parte interna del passaggio. </li><li> Verificare che i motori del robot sono spenti, avviare la routine di calibrazione Phantom e verificare che la lettura di posizione dei motori dalla routine di calibrazione del robot sono stabili. </li><li> Doppia controllare il cavo di collegamento in parallelo e che solo il filtro di grandi dimensioni personalizzate è collegato se la procedura di taratura non può vedere il robot o ci sono grandi variazioni nella lettura del motore. </li><li> Accendere il robot aprendo la porta sul retro della scatola schermatura e premere gli interruttori con un bastone. </li><li> Riavviare la procedura di calibrazione con il effecter fine circa centrato alla fine della guida d&#39;onda conica. Passo attraverso la calibrazione e controllare che la casella di calibrazione box ha il un&#39;adeguata interazione tattile. </li><li> Collegare l&#39;uscita TTL dalla fMRI (connettore BNC) per l&#39;ADC Labjack sul computer di controllo. </li></ol> 4. Il soggetto <ol><li> Preparare il soggetto per un ambiente di alto campo magnetico con i protocolli standard di fMRI. </li><li> Impostare qualsiasi equipaggiamento extra per l&#39;esperimento, ad esempio, il sistema di visualizzazione. Usiamo il NordicNeuro Lab, Vision System Inc. che offre una visualizzazione stereo di visual display, una caratteristica che è particolarmente utile al momento della presentazione degli ambienti virtuali. </li><li> Dopo che il soggetto si sdraia sul tavolo e la bobina testa si trova, regolare la distanza del robot allentando le viti a mano e facendo scorrere la parte superiore del tavolo fino a quando il soggetto può muoversi comodamente. </li><li> Guida alla tabella manualmente premendo sull&#39;estremità del robot, mentre la tabella fMRI si muove dentro e fuori il foro per evitare che le rotelle di oscillazione. Assicurarsi che il effecter fine va nel foro e non prendere sul lato esterno. </li><li> Esegui l&#39;esperimento. </li></ol> 5. Abbattere il setup con due persone A e B <ol><li> Dopo le uscite paziente, rimuovere alla fine esterno del manico lungo e staccare il tavolo robot dalla tabella fMRI annullando il cinturino in velcro. </li><li> Dal robot, annullare il cavo schermato parallelo e scollegare il cavo di alimentazione. </li><li> Un&#39;azienda con la persona l&#39;estremità libera del tavolo robot e la persona B guida la fine robot, spostare il tavolo verso la porta. Alla porta, annullare la pastoia, e ruotare la tavola robot verso la sala. </li><li> Allentare tutte le viti dalla casella schermatura e le due viti dalla piastra di sicurezza e rimuovere il robot. </li></ol> 6. Rappresentante dei risultati: Idealmente, il robot tattili e fMRI non dovrebbe influenzare l&#39;un l&#39;altro. Possiamo dire in linea se il robot viene colpiti dalla fMRI. In generale, se cavo parallelo del robot non è adeguatamente schermato e filtrato, poi la lettura dei motori sarà oscilloscopifine rapidamente. Questo può essere risolto con un doppio controllo della schermatura in alluminio sul cavo, che il nucleo ferroso sia correttamente posizionato sul cavo parallelo vicino il robot, e che l&#39;unico filtro per il robot è il filtro personalizzato sul lato scanner sala del passaggio . Rilevare errori nella fMRI è davvero possibile solo dopo che i dati sono stati ridotti e analizzati, ma una scansione anatomica dovrebbe essere presa presto nello studio e controllato per gli effetti di cerniera o di altri artefatti indicativi di rumore correlato (per esempio rumore picco) 7. Frequentemente, il rumore che questi provengano da metallo a contatto metallo e possono essere ripulite serrando tutte le viti sul tavolo del robot, in particolare le viti di regolazione mano sul lato del tavolo. Dal nostro test il segnale fMRI basale a rumore (SNR) è ~ 380 e con il robot completamente schermati nella stanza che scende ad un ancora ragionevole ~ 330. Se lo scudo non è presente sul robot, poi la goccia SNR ulteriormente ~ 250, e gli effetti del rumore diventare molto significativo. Come mostrato in 4, i 3 gradi di libertà comuni nel centro del manico ha scarso effetto sulla dinamica del robot / interazione mano se non a spostare lontano dal robot. Il giunto nel centro della maniglia agisce come un fulcro e inverte il moto apparente in due delle direzioni (destra-sinistra e alto-basso), ma non il terzo (avanti-indietro). Dato che la Phantom e la mano sono agli estremi opposti della leva come il manico con il suo fulcro nel mezzo, gli utili vengono applicate nel software in ciascuna delle tre direzioni euclidea: guadagna negativa nelle due direzioni controllato dalla snodo e un guadagno positivo nella direzione di scorrimento del giunto. L&#39;effetto netto della maniglia girevole e riproduce l&#39;intero 3 gradi di libertà del robot Phantom, a soli 9 &#39;via. <img src="/files/ftp_upload/3364/3364fig1.jpg" alt="Figura 1"/> Figura 1 L&#39;apparecchio utilizzato per montare il robot tattile per l&#39;uso in ambiente fMRI. Superiore mostra il robot tattile montato nel caso prima di custodia (in alto, a sinistra) e il giunto cardanico / cursore giunto a metà del manico (in alto, a destra). In basso, a sinistra mostra un soggetto nello scanner manipolare la maniglia. Fondo, è proprio un cartone animato della effecter schermatura e fine. <img src="/files/ftp_upload/3364/3364fig2.jpg" alt="Figura 2"/> Figura 2 I risultati del test BIRN per il robot completamente schermati con il movimento. Le tre immagini mostrano con croci sezioni del modello testa sferica, e il basso a destra mostra una vista tridimensionale. I puntini sono bolle in testa il modello statico e sono sempre presenti. La mancanza delle strisce grandi o cerniere indica che il rumore dal robot non correlati.

<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>

Nessun conflitto di interessi dichiarati.

Il robot compatibile fMRI apre nuove possibilità per gli esperimenti in neuroscienze del controllo motorio. La fase più critica nel setup è la schermatura del robot per evitare artefatti nella fMRI, che noi facciamo in due fasi. In primo luogo, lo stesso robot è di circa 9 &#39;dal foro con un lungo, leggero, manico sostenuto nella sua metà con un grado 3 della comune libertà. In secondo luogo, il robot è racchiuso in un 1 / 16 &quot;-1 / 4&quot; scatola di alluminio con una in plastica conica (13 &quot;di diamet...

<table border="1"><tbody><tr><td> Phantom premio 1.5/6dof, modello forza elevata </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

La risonanza magnetica funzionale (fMRI), fornisce un eccellente imaging funzionale del cervello tramite il segnale BOLD 1 con vantaggi tra cui radiazioni non ionizzanti, precisione millimetrica spaziale dei dati anatomici e funzionali 2, e quasi in tempo reale analisi 3. Robot tattile fornire una misurazione precisa e il controllo della posizione e la forza di un cursore in uno spazio ragionevolmente ristretti. Qui combinare queste due tecnologie per consentire gli esperimenti che coinvolgono la precisione di controllo del motore con haptic / interazione ambiente tattili, quali il raggiungimento o afferrare. L&#39;idea di base è quella di collegare un effecter end 8 piedi supportati al centro per il robot 4 che permette al soggetto di utilizzare il robot, ma proteggendolo e mantenendolo fuori della parte più estrema del campo magnetico dalla macchina fMRI (Figura 1 ). Il fantasma Premium 3.0, 6DoF, alta forza di robot (SensAble Technologies, Inc.) è una scelta eccellente per la fornitura di force-feedback in esperimenti di realtà virtuale 5, 6, ma è intrinsecamente non-MR cassetta di sicurezza, introduce rumore significativo al sensibile attrezzature fMRI, e la sua motori elettrici possono essere influenzati dalla fMRI è fortemente variabile campo magnetico. Abbiamo costruito un tavolo e sistema di schermatura che consente al robot di essere tranquillamente introdotti nell&#39;ambiente fMRI e limita sia la degradazione del segnale fMRI dai motori elettrici rumorosi e il degrado delle prestazioni motore elettrico dal campo magnetico variabile forte delle fMRI. Con lo scudo, il rapporto segnale rumore (SNR: media segnale / rumore deviazione standard) della fMRI va da un basale di ~ 380 a ~ 330 e ~ 250 senza la schermatura. Il rumore residuo risulta essere non correlati e non aggiunge artefatti per la fMRI di una sfera di prova (Figura 2). La lunga maniglia rigida permette il posizionamento del robot fuori dalla portata delle parti più fortemente variabili del campo magnetico quindi non c&#39;è alcun effetto significativo della fMRI sul robot. L&#39;effetto della maniglia sulla cinematica del robot è minimo in quanto è leggero (~ 2.6 lbs), ma estremamente rigida 3 / 4 &quot;grafite e ben bilanciata sul giunto 3DoF nel mezzo. Il risultato finale è un fMRI compatibile, sistema aptico con circa 1 piede cubico di spazio di lavoro, e, quando combinato con la realtà virtuale, permette una nuova serie di esperimenti da eseguire in un ambiente naturalistico tra cui fMRI raggiungere, lo spostamento passivo dell&#39;arto e la percezione tattile, l&#39;apprendimento di adattamento in diversi campi di forza , o l&#39;identificazione struttura 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

L&#39;adattamento e l&#39;uso di un robot tattile in un fMRI 3T è descritto.

Adattamento di un robot tattile in un fMRI 3T

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 Views.  University of California. L'adattamento e l'uso di un robot tattile in un fMRI 3T è descritto.

Video: 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.

Fabbricazione di elettrochimica-DNA biosensori per il rilevamento Reagentless di acidi nucleici, proteine ​​e piccole molecole

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. ...

Ricerca

Bioingegneria

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.

Haptic / grafica di riabilitazione: integrazione di un robot in un ambiente di biblioteca virtuale e sua applicazione alla terapia Corsa

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.

La creazione di substrati Patterned bidimensionali per confinamento di proteine ​​e cellule

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).

Estrazione dei lipidi cellulari per analisi mirate Stabile diluizione isotopica cromatografia liquida-spettrometria di massa

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.

Progettazione di un robot bio-sensibile da DNA Origami

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.

Identificazione di ossido di metallo nanoparticelle in campioni istologici da avanzato Mapping Darkfield Microscopy e iperspettrale

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.

Medico-grade di destinazione sterilizzabile per fetoscope ottico Distortion calibrazione Fluid-immersi

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.

Ottimizzare l'utilizzo di un Robot di manipolazione liquido per condurre un alto Throughput avanti chimica genetica schermo di 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 ...

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.

Decellularizzazione di cuore umano intero all'interno di una custodia pressurizzata in un orientamento invertito

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.

Nanoemulsione Tripeptide-stabilizzato di acido oleico

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 ...