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

Мы благодарим доктора Krishanu Ray для акций Drosophila, Tarjani Agarwal для поддержания дрозофилы клетки, Петр Juo для nuIs25 и CGC для C. Элеганс штаммов. SPK сделал jsIs609 в лаборатории Майкла Нонет в. Мы благодарим Arpan Агнихотри (BITS Pilani) за помощь в покадровой визуализации митохондрий перевозки jsIs609 животных в микрофлюидных устройств. Мы благодарны д-р ватсала Thirumalai и Сурия Пракаш за предоставление нам эмбрионов рыбок данио. Мы благодарим доктора Кришны и CIFF в НКО для использования вращающийся диск конфокальной микроскопии при поддержке Департамента Науки и Технологии-центр нанотехнологий (№ SR/55/NM-36-2005). Мы также благодарим Kaustubh Рау, В. Венкатраман и Четана Sachidanand для обсуждения. Эта работа финансировалась DBT пост-докторские стипендии (SM), DST ускоренной схеме (SM) и DBT гранта (SPK). SA была поддержана DST и CSIR гранты SPK.

1. SU8 мастер изготовления <ol><li> Дизайн микрофлюидных структур с использованием Clewin программного обеспечения и распечатать его с помощью плоттера 65024 DPI лазер с минимальным размером особенность 8 мкм на пленке плате. </li><li> Очистите 2 см х 2 см пластины кремния с родным азота в 20% KOH в течение...

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P., Gilday, S. D., Hove, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dose-dependent+effects+of+chemical+immobilization+on+the+heart+rate+of+embryonic+zebrafish.">Dose-dependent effects of chemical immobilization on the heart rate of embryonic zebrafish.</a> Lab. Anim. (NY). 35, 41-47 (2006).</li><li>Pardo-Martin, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-throughput+in+vivo+vertebrate+screening.">High-throughput in vivo vertebrate screening.</a> Nat. Methods. 7, 634-636 (2010).</li><li>Stainier, D. Y., Lee, R. K., Fishman, M. 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PDMS микрофлюидных устройств оптически прозрачным, следовательно, может быть использована для высокого разрешения в естественных изображений любых прозрачных / полупрозрачных моделей организма. Наш дизайн подходит для больших увеличениях пространственно-временных изображений к...

<table border="1"><tbody><tr><td> Название реагента </td><td> Компания </td><td> Номер в каталоге </td><td> Комментарии (по желанию) </td></tr><tr><td> Кремниевые пластины </td><td> Университет пластины </td><td> 150 мм (100) Мех класса SSP Si </td><td></td></tr><tr><td> Clewin программного обеспечения </td><td> WieWeb программного обеспечения </td><td> Версия 2,90 </td><td></td></tr><tr><td> Лазерный плоттер </td><td> Изобразительное изображений линия </td><td> 65024 DPI </td><td></td></tr><tr><td> HMDS </td><td> Sigma-Aldrich </td><td> 440 191-100мл </td><td></td></tr><tr><td> SU8 </td><td> Microchem </td><td> SU8-2025, SU8-2050 </td><td></td></tr><tr><td> Разработчик </td><td> Microchem </td><td> SU8 разработчика </td><td></td></tr><tr><td> Силан </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> УФ-лампы </td><td> Эркер </td><td> 66943 </td><td> 200W Hg Oriel света </td></tr><tr><td> Печь с горячим воздухом </td><td> Ультра инструменты </td><td> На заказ </td><td> Установить при 50 ° C </td></tr><tr><td> Горячая плита </td><td> Оборудование лаборатории МАК </td><td> 3810000 </td><td> <a href="http://www.ika.com" target="_blank">http://www.ika.com</a> </td></tr><tr><td> Плазменный фильтр </td><td> Harrick плазменных </td><td> PDC-32G </td><td></td></tr><tr><td> Прядильщик </td><td> Системы производства полупроводников </td><td> SPIN150-АЭС </td><td> <a href="http://www.SPS-Europe.com" target="_blank">www.SPS-Europe.com</a> </td></tr><tr><td> Скольжения Стеклянная крышка </td><td> Gold Seal </td><td> 22 X 22 мм, толщина № 1 </td><td></td></tr><tr><td>C. Элеганс </td><td> Caenorhabditis генетический центр (CGC) </td><td> e1265, ayIs4 </td><td></td></tr><tr><td> Дрозофила </td><td> Блумингтон </td><td> P {chaGAL4} / CYO, UAS-syt.eGFP </td><td></td></tr><tr><td> Данио рерио </td><td> Индийский дикого типа </td><td> Дикий тип </td><td></td></tr><tr><td> Tygon трубы </td><td> Сигма </td><td> Z279803 </td><td></td></tr><tr><td> Micro иглы </td><td> Сигма </td><td> Z118044 </td><td> Нарезать 1 шт см </td></tr><tr><td> 3-ходовой кран </td><td> Сигма </td><td> S7521 </td><td></td></tr><tr><td> Харрис нокаутер </td><td> Сигма </td><td> Z708631 </td><td></td></tr><tr><td> Сжатого газообразного азота </td><td> Местный поставщик газа </td><td></td><td> Используйте регулятор для контроля давления </td></tr><tr><td> Стерео микроскоп &lt;/ TD&gt; <td> Nikon </td><td> SMZ645 </td><td></td></tr><tr><td> Конфокальной микроскопии </td><td> Андор и Olympus </td><td> Yokogawa вращающийся диск конфокальной микроскопии </td><td></td></tr><tr><td> ImageJ </td><td> Национальные институты здоровья </td><td> <a href="http://www.rsbweb.nih.gov/ij" target="_blank">www.rsbweb.nih.gov / ц</a> </td><td> Java на основе программы обработки изображений </td></tr></tbody></table>

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

Micro fabricated fluidic devices provide an accessible micro-environment for in vivo studies on small organisms. Simple fabrication processes are available for microfluidic devices using soft lithography techniques 1-3. Microfluidic devices have been used for sub-cellular imaging 4,5, in vivo laser microsurgery 2,6 and cellular imaging 4,7. In vivo imaging requires immobilization of organisms. This has been achieved using suction 5,8, tapered channels 6,7,9, deformable membranes 2-4,10, suction with additional cooling 5, anesthetic gas 11, temperature sensitive gels 12, cyanoacrylate glue 13 and anesthetics such as levamisole 14,15. Commonly used anesthetics influence synaptic transmission 16,17 and are known to have detrimental effects on sub-cellular neuronal transport 4. In this study we demonstrate a membrane based poly-dimethyl-siloxane (PDMS) device that allows anesthetic free immobilization of intact genetic model organisms such as Caenorhabditis elegans (C. elegans), Drosophila larvae and zebrafish larvae. These model organisms are suitable for in vivo studies in microfluidic devices because of their small diameters and optically transparent or translucent bodies. Body diameters range from ~10 &mu;m to ~800 &mu;m for early larval stages of C. elegans and zebrafish larvae and require microfluidic devices of different sizes to achieve complete immobilization for high resolution time-lapse imaging. These organisms are immobilized using pressure applied by compressed nitrogen gas through a liquid column and imaged using an inverted microscope. Animals released from the trap return to normal locomotion within 10 min.
We demonstrate four applications of time-lapse imaging in C. elegans namely, imaging mitochondrial transport in neurons, pre-synaptic vesicle transport in a transport-defective mutant, glutamate receptor transport and Q neuroblast cell division. Data obtained from such movies show that microfluidic immobilization is a useful and accurate means of acquiring in vivo data of cellular and sub-cellular events when compared to anesthetized animals (Figure 1J and 3C-F 4).
Device dimensions were altered to allow time-lapse imaging of different stages of C. elegans, first instar Drosophila larvae and zebrafish larvae. Transport of vesicles marked with synaptotagmin tagged with GFP (syt.eGFP) in sensory neurons shows directed motion of synaptic vesicle markers expressed in cholinergic sensory neurons in intact first instar Drosophila larvae. A similar device has been used to carry out time-lapse imaging of heartbeat in ~30 hr post fertilization (hpf) zebrafish larvae. These data show that the simple devices we have developed can be applied to a variety of model systems to study several cell biological and developmental phenomena in vivo.

Иммобилизации устройства двухслойной блоке PDMS изготовлены путем склеивания двух слоев: слоя потока (Layer 1) и контроля слой (Layer 2), как показано на рисунке 1. Основная ловушка соединена с цилиндром азота через регулятор и 3-ходовой запорный кран применять необходимые (3-14 бар) давле...

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

Простой микрофлюидных устройство было разработано для выполнения анестетика бесплатно<em&gt; В естественных условиях</em&gt; Визуализации<em&gt; C. Элеганс</em&gt;, Неповрежденные<em&gt; Drosophila</em&gt; Личинок и рыбок данио личинки. Устройство используется деформируемый мембраны PDMS, чтобы обездвижить этих модельных организмов в целях выполнения промежуток времени визуализации многочисленных процессов, таких как сердцебиение, деление клеток и субклеточных нейронов транспорта. Мы демонстрируем использование этого устройства и приведены примеры различных типов данных, собранных из различных модельных системах.

Простой микрофлюидных устройства для<em&gt; В естественных условиях</em&gt; Визуализация<em&gt; C. Элеганс</em&gt;<em&gt; Drosophila</em&gt; И данио рерио

Micro fabricated fluidic devices provide an accessible micro-environment for in vivo studies on small organisms. Simple fabrication processes are ...

Research

JoVE Journal

Bioengineering

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

Separating Beads and Cells in Multi-channel Microfluidic Devices Using Dielectrophoresis and Laminar Flow

Разделение Бусы и клетки в Многоканальные Устройства микрожидкостных Использование Диэлектрофорез и ламинарного потока

Microfluidic devices have advanced cell studies by providing a dynamic fluidic environment on the scale of the cell for studying, manipulating, sorting ...

Viral Nanoparticles for In vivo Tumor Imaging

Viral Nanoparticles for In vivo Tumor Imaging

Вирусный наночастиц для<em&gt; В естественных условиях</em&gt; Изображения опухоли

The use of nanomaterials has the potential to revolutionize materials science and medicine. Currently, a number of different nanoparticles are being ...

A Microfluidic Technique to Probe Cell Deformability

Микрожидкостных Техника для исследования клеток деформируемости

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

Слой за слоем Коллаген осаждения в микрофлюидных Устройства для стабилизации Microtissue

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

Электротаксис Исследования клетки легких рака с использованием многоканального Dual-электрического поля микрофлюидных Chip

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

Multi-step Variable Height Photolithography for Valved Multilayer Microfluidic Devices

Многоступенчатая переменной высоты фотолитографии для Valved Многослойные микрожидком устройств

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

Изготовление Трехмерная бумаги на основе микрожидком Устройства для иммуноанализа

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

Полидиметилсилоксан-поликарбонатные микрожидком Устройства для миграции клеток исследований при Перпендикулярного химической и кислорода Градиентов

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

Простой и масштабируемый способ изготовления для органических электронных устройств по текстилю

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

A Microfluidic Platform for Longitudinal Imaging in Caenorhabditis elegans

A Microfluidic Platform for Longitudinal Imaging in Caenorhabditis elegans

Microfluidic платформа для продольной Imaging в Caenorhabditis elegans

In the last decade, microfluidic techniques have been applied to study small animals, including the nematode Caenorhabditis elegans, and have proved ...