Name: simple microfluidic devices for in vivo imaging of c. elegans, drosophila and zebrafish
Uploaded: 2012-09-30T14:00:00
Description: Watch this Scientific Journal Video about Simple Microfluidic Devices for in vivo Imaging of C. elegans, Drosophila and Zebrafish at JoVE.com

Ringraziamo il Dott. Krishanu Ray per gli stock di Drosophila, Tarjani Agarwal per il mantenimento di una gabbia di Drosophila, Peter Juo per nuIs25 e CGC per C. elegans ceppi. SPK fatto jsIs609 nel laboratorio di Michael Nonet. Ringraziamo Arpan Agnihotri (BITS Pilani) per il suo aiuto in time-lapse imaging di trasporto dei mitocondri jsIs609 animali in dispositivi microfluidici. Siamo grati al Dr. Vatsala Thirumalai e Surya Prakash per averci fornito embrioni di zebrafish. Ringraziamo il Dott. Krishna e CIFF a BCN per l&#39;uso del microscopio confocale disco rotante sostenuto dal Dipartimento di Scienze e Tecnologie-Centro per le Nanotecnologie (n. SR/55/NM-36-2005). Ringraziamo anche Kaustubh Rau, V. Venkatraman e Chetana Sachidanand per le discussioni. Questo lavoro è stato finanziato dal DBT post-dottorato (SM), DST accelerata regime (SM) e una borsa di studio a DBT (SPK). SA è stata sostenuta da sovvenzioni DST e CSIR a SPK.

1. SU8 Maestro Fabrication <ol><li> Progettare le strutture microfluidica utilizzando software Clewin e stamparlo con 65.024 plotter DPI laser con dimensione minima di 8 micron su pellicola circuito. </li><li> Pulire 2 cm x 2 centimetri wafer di silicio con ossido nativo in KOH al 20% per 1 min e risciacquo in acqua deionizzata; wafer uno strato per ogni flusso e il suo livello di controllo corrispondente. </li><li> Asciugare i pezzi con gas azoto e disidratare su una piastra calda a 120 ° C per 4 h. Permettono i pez...

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PDMS dispositivi microfluidici sono otticamente trasparente, pertanto può essere utilizzata per alta risoluzione immagini in vivo di qualsiasi trasparente / traslucido organismo modello. Il nostro design è adatto per l&#39;alta ingrandimento spazio-temporale di immagini di eventi cellulari e sub-cellulari in intatti animali vivi. Microfabrication con tecniche di litografia soft permette una facile manipolazione delle dimensioni del dispositivo disponibili per varie dimensioni di organismi modello. Dispositivi...

<table border="1"><tbody><tr><td> Nome del reagente </td><td> Azienda </td><td> Numero di catalogo </td><td> Commenti (opzionale) </td></tr><tr><td> Wafer di silicio </td><td> Università di wafer </td><td> 150 mm (100) Mech Grado SSP Si </td><td></td></tr><tr><td> Clewin Software </td><td> WieWeb software </td><td> Versione 2.90 </td><td></td></tr><tr><td> Plotter laser </td><td> Linea Imaging fine </td><td> 65024 DPI </td><td></td></tr><tr><td> HMDS </td><td> Sigma-Aldrich </td><td> 440191-100ML </td><td></td></tr><tr><td> SU8 </td><td> Microchem </td><td> SU8-2025, SU8-2050 </td><td></td></tr><tr><td> Sviluppatore </td><td> Microchem </td><td> SU8 Developer </td><td></td></tr><tr><td> Silano </td><td> Sigma-Aldrich </td><td> 448931-10G </td><td></td></tr><tr><td> PDMS </td><td> Dow corning </td><td> Sylgard 184 </td><td></td></tr><tr><td> Lampada UV </td><td> Finestra sporgente </td><td> 66943 </td><td> 200W Hg Oriel Luce </td></tr><tr><td> Stufa ad aria calda </td><td> Strumenti Ultra </td><td> Una soluzione su misura </td><td> A 50 ° C </td></tr><tr><td> Placca di cottura </td><td> IKA Apparecchiature di laboratorio </td><td> 3810000 </td><td> <a href="http://www.ika.com" target="_blank">http://www.ika.com</a> </td></tr><tr><td> Plasma cleaner </td><td> Harrick Plasma </td><td> PDC-32G </td><td></td></tr><tr><td> Spinner </td><td> Semiconduttore e Sistemi di Produzione </td><td> SPIN150-NPP </td><td> <a href="http://www.SPS-Europe.com" target="_blank">www.sps-Europe.com</a> </td></tr><tr><td> Vetro fodera </td><td> Gold Seal </td><td> 22 X 22 mm, spessore No. 1 </td><td></td></tr><tr><td>C. elegans </td><td> Caenorhabditis Genetics Center (CGC) </td><td> e1265, ayIs4 </td><td></td></tr><tr><td> Drosophila </td><td> Bloomington </td><td> P {} chaGAL4 / cyo, UAS-syt.eGFP </td><td></td></tr><tr><td> Zebrafish </td><td> Indiano wild type </td><td> Tipo selvatico </td><td></td></tr><tr><td> Tygon Tubo </td><td> Sigma </td><td> Z279803 </td><td></td></tr><tr><td> Micro ago </td><td> Sigma </td><td> Z118044 </td><td> Tagliare a pezzi di cm 1 </td></tr><tr><td> Rubinetto a 3 vie </td><td> Sigma </td><td> S7521 </td><td></td></tr><tr><td> Harris puncher </td><td> Sigma </td><td> Z708631 </td><td></td></tr><tr><td> Gas azoto compresso </td><td> Gas locale </td><td></td><td> Utilizzare un regolatore per il controllo della pressione </td></tr><tr><td> Microscopio Stereo &lt;/ Td&gt; <td> Nikon </td><td> SMZ645 </td><td></td></tr><tr><td> Microscopio confocale </td><td> Andor &amp; Olympus </td><td> Yokogawa filatura disco microscopio confocale </td><td></td></tr><tr><td> ImageJ </td><td> National Institutes of Health </td><td> <a href="http://www.rsbweb.nih.gov/ij" target="_blank">www.rsbweb.nih.gov / IJ</a> </td><td> Basato su Java programma di elaborazione delle immagini </td></tr></tbody></table>

simple microfluidic devices for in vivo imaging of c. elegans, drosophila and zebrafish

Micro dispositivi fabbricati fluidici fornire un micro-ambiente accessibile per gli studi in vivo su piccoli organismi. Processi di fabbricazione semplici sono disponibili per i dispositivi microfluidici che utilizzano tecniche di litografia morbidi 1-3. Dispositivi microfluidici sono stati utilizzati per la sub-cellulare di imaging 4,5, in vivo laser microchirurgia 2,6 e 4,7 di imaging cellulare. In vivo imaging richiede immobilizzazione di microrganismi. Questo è stato ottenuto utilizzando aspirazione 5,8, 6,7,9 canali conici, membrane deformabili 2-4,10, aspirazione con raffreddamento supplementare 5, gas anestetico 11, gel sensibili alla temperatura 12, 13 e colla cianoacrilato anestetici come levamisole 14 , 15. Anestetici comunemente usati influenzare la trasmissione sinaptica 16,17 e sono noti per avere effetti nocivi sulla sub-cellulare 4 Trasporti neuronale. In questa study si dimostra una membrana a base di poli-dimetil-silossano (PDMS), dispositivo che permette di immobilizzazione senza anestetico intatte organismi modello genetiche come Caenorhabditis elegans (C. elegans), larve di Drosophila e larve di zebrafish. Questi organismi modello sono adatti per gli studi in vivo in dispositivi microfluidici causa dei loro diametri piccoli e organismi otticamente trasparenti o traslucide. Diametri del corpo vanno da ~ 10 micron a ~ 800 micron per i primi stadi larvali di C. elegans e zebrafish larve e richiedono dispositivi microfluidici di diverse dimensioni per ottenere l &#39;immobilizzazione completa ad alta risoluzione time-lapse imaging. Questi organismi sono immobilizzati mediante pressione esercitata dal gas azoto compresso attraverso una colonna di liquido e ripreso utilizzando un microscopio invertito. Animali rilasciati dalla trappola tornare alla normale locomozione entro 10 min. Dimostriamo quattro applicazioni di time-lapse imaging in C. elegansvale a dire, il trasporto di imaging mitocondriale nei neuroni, pre-sinaptico di trasporto delle vescicole in un trasporto difettoso-mutante, il trasporto recettori del glutammato e la divisione neuroblasti Q cellulare. I dati ottenuti da questi film mostrano che l&#39;immobilizzazione microfluidica è un mezzo utile e preciso di acquisire dati in vivo di eventi cellulari e sub-cellulari rispetto agli animali anestetizzati (Figura 1J e 3C-F 4). Dimensioni del dispositivo sono state modificate per consentire time-lapse imaging di diverse fasi di C. elegans, Drosophila primo stadio le larve e larve di zebrafish. Trasporto di vescicole contrassegnati con synaptotagmin contrassegnati con GFP (syt.eGFP) nei neuroni sensoriali mostra movimento diretto di marcatori delle vescicole sinaptiche espressi nei neuroni colinergici sensoriali intatte prima larve instar Drosophila. Un dispositivo simile è stato utilizzato per effettuare time-lapse imaging del battito cardiaco in ~ 30 ore dopo la fecondazione (HPF)Zebrafish larve. Questi dati mostrano che i dispositivi semplici che abbiamo sviluppato può essere applicato ad una varietà di sistemi modello per lo studio di fenomeni biologici diversi cellulari e di sviluppo in vivo.

Il dispositivo di immobilizzazione è un blocco bistrato PDMS fabbricato legando due strati: uno strato di flusso (Layer 1) e uno strato di controllo (Layer 2) come mostrato in Figura 1. La trappola principale è collegato ad una bombola di gas azoto attraverso un regolatore ed un rubinetto di arresto a 3 vie per applicare necessaria (3-14 psi) di pressione sulla membrana attraverso una colonna di liquido (Figura 1A). La membrana deviato immobilizza C. larve elegans, Drosop...

Watch this Scientific Journal Video about Simple Microfluidic Devices for in vivo Imaging of C. elegans, Drosophila and Zebrafish at JoVE.com

Simple Microfluidic Devices for in vivo Imaging of C. elegans, Drosophila and Zebrafish

Un semplice dispositivo microfluidico è stato sviluppato per eseguire anestetico libera<em&gt; In vivo</em&gt; Imaging<em&gt; C. elegans</em&gt;, Intatto<em&gt; Drosophila</em&gt; Larve e larve di zebrafish. Il dispositivo utilizza una membrana deformabile PDMS per immobilizzare questi organismi modello per eseguire lapse imaging tempo di numerosi processi come il battito cardiaco, la divisione cellulare e subcellulare trasporto neuronale. Abbiamo illustrato l&#39;utilizzo di questo dispositivo e mostrano esempi di diversi tipi di dati raccolti da diversi sistemi modello.

Semplici dispositivi microfluidici per<em&gt; In vivo</em&gt; Imaging di<em&gt; C. elegans</em&gt;,<em&gt; Drosophila</em&gt; E Zebrafish

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Simple Microfluidic Devices for in vivo Imaging of C. elegans, Drosophila and Zebrafish

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A Microfluidic Technique to Probe Cell Deformability

Una tecnica Microfluidic per sondare deformabilità cellulare

Here we detail the design, fabrication, and use of a microfluidic device to evaluate the deformability of a large number of individual cells in an ...

Layer-by-layer Collagen Deposition in Microfluidic Devices for Microtissue Stabilization

Layer-by-layer collagene Deposizione nel Microfluidic Dispositivi per microtessuto Stabilizzazione

Although microfluidics provides exquisite control of the cellular microenvironment, culturing cells within microfluidic devices can be challenging. 3D ...

Electrotaxis Studies of Lung Cancer Cells using a Multichannel Dual-electric-field Microfluidic Chip

Electrotaxis Studi di polmone cellule tumorali usando un Multicanale Dual-campo elettrico Microfluidic Chip

The behavior of directional cell migration under a direct current electric-field (dcEF) is referred to as electrotaxis. The significant role of ...

Using Adhesive Patterning to Construct 3D Paper Microfluidic Devices

Utilizzando adesivo Patterning per costruire dispositivi microfluidici di carta 3D

We demonstrate the use of patterned aerosol adhesives to construct both planar and nonplanar 3D paper microfluidic devices. By spraying an aerosol ...

Multi-step Variable Height Photolithography for Valved Multilayer Microfluidic Devices

Multi-step altezza variabile fotolitografia per Valvolati multistrato microfluidici Devices

Microfluidic systems have enabled powerful new approaches to high-throughput biochemical and biological analysis. However, there remains a barrier to ...

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

Fabbricazione di dispositivi microfluidici basati su carta tridimensionale per gli immunodosaggi

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

Polydimethylsiloxane-polycarbonate Microfluidic Devices for Cell Migration Studies Under Perpendicular Chemical and Oxygen Gradients

Polidimetilsilossano-policarbonato dispositivi microfluidici per Cell Migration Studies Sotto perpendicolare chimica e gradienti di ossigeno

This paper reports a microfluidic device made of polydimethylsiloxane (PDMS) with an embedded polycarbonate (PC) thin film to study cell migration under ...

A Simple and Scalable Fabrication Method for Organic Electronic Devices on Textiles

Un metodo semplice e scalabile di fabbricazione per dispositivi elettronici organici sui tessili

Today, wearable electronics devices combine a large variety of functional, stretchable, and flexible technologies. However, in many cases, these devices ...