이 작품은 Adeola Adewola, NIH / NCRR (U42RR023245) 호세 Oberholzer하고, 시카고 당뇨병 프로젝트 AAUW 국제 친교에 의해 지원되었다.

3 층 microfluidic의 perifusion 장치의 A.의 Microfabrication 하단 그럼 마스터 프로토콜 (150 μm의 깊은 우물) <ol><li> 필요한 경우 면도날을 사용하여 웨이퍼를 청소합니다. 아세톤, 메탄올과 IPA로 닦으십시오. 30 S. 50 왓츠에서 플라즈마 처리를 수행 </li><li> 스핀 SU8 - 100 @ 2000 RPM. [1 단계 : 500 RPM, 10 S, 100 RPM / s의 2 단계 : 2000 RPM, 30 S, 300 RPM / S]. [참고 : SU8 스피닝 후 핀셋으로 웨이퍼를 보유하지] </li><li> 소프트 베이킹 65 웨이퍼 ° C 20 분 및 95 °에서 50 분 C. </li><li> 웨이퍼는 원하는 마스크를 통해 자외선에 노출됩니다. 150 μm의 높이에 대한 선량 650 MJ / cm 2입니다. </li><li> 포스트 노출은 65에서 웨이퍼를 구워 ° 1 분 및 95에서 C ° 12 분 C. </li><li> 15 분 SU8 개발자의 웨이퍼를 개발. </li></ol> Microchannel 마스터 프로토콜 (500 μm의 깊은 우물) <ol><li> 필요한 경우 면도날을 사용하여 웨이퍼를 청소합니다. 아세톤, 메탄올과 IPA로 닦으십시오. 30 S. 50 왓츠에서 플라즈마 처리를 수행 </li><li> 스핀 SU8 - 2150 @ 1000 RPM. [1 단계 : 500 RPM, 10 S, 100 RPM / s의 2 단계 : 1000 RPM, 30 S, 300 RPM / S]. </li><li> 소프트 베이킹 65 웨이퍼 ° C 15 분 및 95 °에서 2 시간 30 분 C. </li><li> 웨이퍼는 원하는 마스크를 통해 자외선에 노출됩니다. 650 μm의 높이에 대한 노출 선량은 685 MJ / cm 2입니다. </li><li> 포스트 노출은 65에서 웨이퍼를 구워 ° 5 분 및 95에서 C ° ~ 35 분 C. </li><li> 20-30 분 SU8 개발자의 웨이퍼를 개발. </li></ol> PDMS 솔루션 준비 <ol><li> Polydimethylsiloxane (PDMS) 솔루션은 철저하게 표준 Sylgard 184 키트의 치료 요원 1 일부로 실리콘 탄성체의 10 부분을 혼합하여 준비가되어 있습니다. </li><li> 혼합 과정에서 PDMS 솔루션에서 생성된 거품은 진공 건조기를 사용하여 제거됩니다. </li><li> 거품없는 PDMS 솔루션은 천천히 SU8 마스터와 세 번째 레이어에 빈 페트리 접시에 적절합니다. </li><li> 뜨거운 접시의 온도가 75로 설정되어 ° C와 PDMS는 2 시간이 온도에서 치료됩니다 </li><li> 인레츠, 콘센트 및 교환 우물은 적절한 크기의 구멍 구멍을 뚫는를 사용하여 밖으로 펀치입니다. </li><li> 레이어는 휴대용 플라즈마 장치를 사용하여 유리 슬라이드 (크기 0.1 ㎜)에 함께 결합하고 있습니다. </li></ol> B. 실험 설정 <ol><li> 마이크로 장치를 통해 흐름을 70 % 에탄올은 소독 수 있습니다. 흐름 DI 물 에탄올을 씻어. microchannel 벽에 인슐린 이외의 구체적인 흡착을 방지하기 위해 장치를 통해 0.5 % BSA의 50 ML을 Perfuse. </li><li> 포도당 램프 기울기와 펌프는 그라디언트가 안정되어 있는지 확인하기 위해 테스트하는 주사기와 통신 LabView 소프트웨어에 의해 생성된 기타 관련 그라디언트. </li><li> 25-30 쥐 아일 레츠가 37 ° C에서 30 분 5 μm의의 Fura-2/AM (칼슘 표시기, 분자 프로브, CA) 및 2.5 μm의의 Rhodamine 123 (Rh123, mitochondrial 가능성 표시기, 시그마, MO)와 incubated되었습니다 이 MM 포도당을 포함한 크렙스 - 링어 버퍼 (KRB) </li><li> 마우스 아일 레츠는 신중하게 유입 포트를 통해 microfluidic perifusion 장치로 pipetted 있습니다. </li><li> microfluidic 네트워크는 다음 Tygon 관 그리고 Y - 커넥터 및 분획 수집기에 콘센트를 사용하여 주사기 펌프에 입구를 연결하여 설정합니다. 장치는 현미경에서 가열 단계 (37 ° C)에 자리하고 유입 튜브는 37 챔버 및 솔루션의 온도를 유지하기 위해 열판에서 가열 ° C. </li><li> 즉시 설치 후, 쥐 아일 레츠는 KRB 그런 다음 10 분 25 분 포도당 램프 (2mM 25mM) 2 MM 포도당이 포함된 함께 perifused 있습니다. </li><li> 시간 경과 이미지 SimplePCI 소프트웨어에 의해 매 15 s를 수집하고 분석합니다. perifusate 또한 엘리사 키트를 사용하여 인슐린 분비를 분석하기 위해 분획 수집기를 사용하여 매 순간을 수집합니다. </li></ol> 대표 결과 마우스 아일 레츠는 2-25 MM 포도당의 선형 그라디언트로 perifused되었습니다. 그림 1에 표시된, 칼슘 유입 및 인슐린 분비는 6 MM 포도당에 해당하는 perifusion 13 분 후에 실행됩니다. Mitochondrial 잠재력의 변화는 약 11 분, 이전에 예상대로 볼 수 있습니다. 이 데이터는 아일릿 생리 특징이 microfluidic 네트워크를 사용하는 장점을 보여줍니다. <img src="/files/ftp_upload/1649/1649fig1.jpg" alt="그림 1" /> 그림 1. 세포 자극을 아일릿하는 생리 반응.

<ol>
	<li>Mohammed, J. S., Wang, Y., Harvat, T. A., Oberholzer, J., Eddington, D. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfluidic+device+for+multimodal+characterization+of+pancreatic+islets.">Microfluidic device for multimodal characterization of pancreatic islets.</a> Lab Chip. 9, 97-106 (2009).</li></ol>

전통 아일릿 perifusion 시스템 (macroscale 및 microscale) 설정 및 디자인, 높은 기술적 요구 사항과 시스템의 사용자 규정된 화학 그라디언트를 만들 어려움 복잡 등 일부 제한이 있습니다. 여기에서 설명한 microfluidic perifusion 시스템 설계 및 제조의 간단한 기하학 이러한 한계를 극복. 더 중요한 것은,이 시스템은 아일릿 생리 연구를 수있는 유일한 도구로 제공하는 형광 이미징 접근 방식과 통합될 수 있...

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>60ml Syringes</td>
<td>&nbsp;</td>
<td>BD</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Fura-2 fluorescence dye</td>
<td>&nbsp;</td>
<td>Molecular Probes Inc.</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Rhodamine123 Fluorescence dye</td>
<td>&nbsp;</td>
<td>Molecular Probes Inc.</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Glucose</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Bovine Serum Albumin</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>30" Silicone tubings</td>
<td>&nbsp;</td>
<td>Cole Palmer</td>
<td>&nbsp;</td>
<td>1/16 x 1/8in</td>
</tr>
<tr>
<td>1.5ml Eppendorf tubes</td>
<td>&nbsp;</td>
<td>Fisher</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Y-connectors</td>
<td>&nbsp;</td>
<td>Cole Palmer</td>
<td>&nbsp;</td>
<td>1/16&#x201D; &amp; 4mm</td>
</tr>
<tr>
<td>Syringe connectors</td>
<td>&nbsp;</td>
<td>Cole Palmer</td>
<td>&nbsp;</td>
<td>female luer plug 1/16&#x201D;</td>
</tr>
<tr>
<td>Straight connectors</td>
<td>&nbsp;</td>
<td>Cole Palmer</td>
<td>&nbsp;</td>
<td>1/16&#x201D;</td>
</tr>
<tr>
<td>Elbow connector</td>
<td>&nbsp;</td>
<td>Cole Palmer</td>
<td>&nbsp;</td>
<td>1/16&#x201D;</td>
</tr>
<tr>
<td>Havard syringe pump</td>
<td>&nbsp;</td>
<td>Harvard Apparatus</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Perifusion device</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Hot plate</td>
<td>&nbsp;</td>
<td>PMC</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Thermometer</td>
<td>&nbsp;</td>
<td>Omega Engineering Inc.</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Fraction collector</td>
<td>&nbsp;</td>
<td>Gibson</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Pippettor</td>
<td>&nbsp;</td>
<td>Fisher</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Inverted epiflorescence microscope</td>
<td>&nbsp;</td>
<td>Olympus</td>
<td>IX71</td>
<td>&nbsp;</td>
<tr>
<td>Charge-coupled device</td>
<td>&nbsp;</td>
<td>QImaging</td>
<td>&nbsp;</td>
<td>Retiga-SRV, Fast 1394</td>
</tr>		</tbody>
		</table>
		</div>

a multi-parametric islet perifusion system within a microfluidic perifusion device

microfluidic 아일릿 perifusion 장치가 여러 아일 레츠와 칼슘 유입과 mitochondrial 잠재적인 변화 동시 형광 이미지의 역동적인 인슐린 분비의 평가를 위해 개발되었습니다. 이 장치는 세 계층으로 구성되어 있습니다 : 첫 번째 레이어는 아일 레츠의 노출 표면적을 흐름과 극대화에 노출된 상태에서 아일 레츠를 무력화하는 데 도움이 microscale 웰스 (500 μm의 직경 150 μm의 깊이)의 배열을 포함, 두 번째 레이어는 원형을 포함 perifusion 챔버 (깊이 3mm, 7mm 직경) 및 세 번째 계층은 입구 - 혼합 채널을 포함하는 팬들 아웃하기 전에 최적화에 대한 perifusion 챔버에 주입 (폭 2mm, 길이 19mm, 높이 500 μm의) 혼합 효율 전에 perifusion 챔버를 입력합니다. 선형, 종 모양 등 다양한 포도당 그라디언트, 그리고 광장 형태의 생성도 microfluidic perifusion 네트워크에 만들 수 있으며, 증명합니다.

Watch this Scientific Journal Video about A Multi-Parametric Islet Perifusion System within a Microfluidic Perifusion Device at JoVE.com

A Multi-Parametric Islet Perifusion System within a Microfluidic Perifusion Device

microfluidic 아일릿 perifusion 장치가 여러 아일 레츠와 칼슘 유입과 mitochondrial 잠재적인 변화 동시 형광 이미지의 역동적인 인슐린 분비의 평가를 위해 개발되었습니다.

Microfluidic Perifusion 장치 내에서 멀티 파라메 트릭 아일릿 Perifusion 시스템

A microfluidic islet perifusion device was developed for the assessment of dynamic insulin secretion of multiple islets and simultaneous fluorescence ...

a-multi-parametric-islet-perifusion-system-within-microfluidic

Research

JoVE Journal

Biology

11.8K 조회수.  University of Illinois, Chicago. microfluidic 아일릿 perifusion 장치가 여러 아일 레츠와 칼슘 유입과 mitochondrial 잠재적인 변화 동시 형광 이미지의 역동적인 인슐린 분비의 평가를 위해 개발되었습니다.

비디오: A Multi-Parametric Islet Perifusion System within a Microfluidic Perifusion Device

Murine Pancreatic Islet Isolation

Murine 췌장 아일릿 격리

연구

생물학

Fabrication of a Microfluidic Device for the Compartmentalization of Neuron Soma and Axons

In this video, we demonstrate the technique of soft lithography with polydimethyl siloxane (PDMS) which we use to fabricate a microfluidic device for culturing neurons. Previously, a silicon wafer was patterned with the design for the neuron microfluidic device using SU-8 and photolithography to create a master mold, or what we simply refer to as a "master". Next, we pour the silicon polymer PDMS on top of the master which is then cured by heating the PDMS to 80&deg;C for 1 hour. The PDMS forms a negative mold of the device. The PDMS is then carefully cut and lifted away from the master. Holes are punched where the reservoirs will be and the excess PDMS trimmed away from the device. Nitrogen is used to blow away any excess debris from the device. At this point the devices are now ready for use and can either bonded to corning No. 1 cover glass with a plasma sterilizer/cleaner or can be reversibly bound to the cover glass by simply placing the device on top of the cover glass. The reversible bonding of the device to glass is covered in a separate video and requires first that the device be sterilized either with 70% ethanol or by autoclaving. Plasma treating sterilizes the devices so no further treatment is necessary. It is, however, important, when plasma-treating the devices, to add liquid to the devices within 10 minutes of the plasma treatment while the surfaces are still hydrophilic. Waiting longer than 10 minutes to add liquid to the device makes it difficult for the liquid to enter the device. The neuron devices are typically plasma-bound to cover glass and 0.5 mg/ml poly-L-lysine (PLL) in pH 8.5 borate buffer is immediately added to the device. After a minimum of 3 hours incubating with PLL, the devices are washed with dH2O water a minimum of 3 times with at least 15 minutes between each wash. Next, the water is removed and fresh media is added to the device. At this point the device is ready for use. It is important to remember at this point to never remove all the media from the device. Always leave media in the main channel.

In this video we demonstrate the technique of soft lithography with polydimethyl siloxane (PDMS) which we use to farbricate a microfluidic device for culturing neurons.

신경 소마와 Axons의 Compartmentalization위한 Microfluidic 장치의 제작

In this video, we demonstrate the technique of soft lithography with polydimethyl siloxane (PDMS) which we use to fabricate a microfluidic device for ...

A Microfluidic Device with Groove Patterns for Studying Cellular Behavior

We describe a microfluidic device with microgrooved patterns for studying cellular behavior. This microfluidic platform consists of a top fluidic channel and a bottom microgrooved substrate. To fabricate the microgrooved channels, a top poly(dimethylsiloxane) (PDMS) mold containing the impression of the microfluidic channels was aligned and bonded to a microgrooved substrate. Using this device, mouse fibroblast cells were immobilized and patterned within microgrooved substrates (25, 50, 75, and 100 &mu;m wide). To study apoptosis in a microfluidic device, media containing hydrogen peroxide, Annexin V, and propidium iodide was perfused into the fluidic channel for 2 hours. We found that cells exposed to the oxidative stress became apoptotic. These apoptotic cells were confirmed by Annexin V that bound to phosphatidylserine at the outer leaflet of the plasma membrane during the apoptosis process. Using this microfluidic device with microgrooved patterns, the apoptosis process was observed in real-time and analyzed by using an inverted microscope containing an incubation chamber (37&deg;C, 5% CO2). Therefore, this microfluidic device incorporated with microgrooved substrates could be useful for studying the cellular behavior and performing high-throughput drug screening.

We describe a protocol for the fabrication of microfluidic devices that can enable cell capture and culture. In this approach patterned microstructures such as grooves within microfluidic channels are used to create low shear stress regions within which cell can dock.

세포 행동 유학을위한 그루브 패턴 Microfluidic 장치

We describe a microfluidic device with microgrooved patterns for studying cellular behavior.  This microfluidic platform consists of a top fluidic channel ...

A Gradient-generating Microfluidic Device for Cell Biology

 The fabrication and operation of a gradient-generating microfluidic device for studying cellular behavior is described.  A microfluidic platform is an enabling experimental tool, because it can precisely manipulate fluid flows, enable high-throughput experiments, and generate stable soluble concentration gradients.  Compared to conventional gradient generators, poly(dimethylsiloxane) (PDMS)-based  microfluidic devices can generate stable concentration gradients of growth factors with well-defined profiles.  Here, we developed simple gradient-generating microfluidic devices with three separate inlets.  Three microchannels combined into one microchannel to generate concentration gradients.  The stability and shape of growth factor gradients were confirmed by fluorescein isothyiocyanate (FITC)-dextran with a molecular weight similar to epidermal growth factor (EGF).  Using this microfluidic device, we demonstrated that fibroblasts exposed to concentration gradients of EGF migrated toward higher concentrations.  The directional orientation of cell migration and motility of migrating cells were quantitatively assessed by cell tracking analysis.  Thus, this gradient-generating microfluidic device might be useful for studying and analyzing the behavior of migrating cells.

We describe a protocol for the microfabrication of the gradient-generating microfluidic device that can generate spatial and temporal gradients in well-defined microenvironment. In this approach, the gradient-generating microfluidic device can be used to study directed cell migration, embryogenesis, wound healing, and cancer metastasis.

세포 생물학을위한 그라디언트 생성 Microfluidic 장치

 The fabrication and operation of a gradient-generating microfluidic device for studying cellular behavior is described.  A microfluidic platform is an ...

Preparing E18 Cortical Rat Neurons for Compartmentalization in a Microfluidic Device

In this video, we demonstrate the preparation of E18 cortical rat neurons. E18 cortical rat neurons are obtained from E18 fetal rat cortex previously dissected and prepared. The E18 cortex is, upon dissection, immediately dissociated into individual neurons. It is possible to store E18 cortex in Hibernate E buffer containing B27 at 4&deg;C for up to a week before the dissociation is performed. However, there will be a drop in cell viability. Typically we obtain our E18 Cortex fresh. It is transported to the lab in ice cold Calcium free Magnesium free dissection buffer (CMFM). Upon arrival, trypsin is added to the cortex to a final concentration of 0.125%. The cortex is then incubated at 37&deg;C for 8 minutes. DMEM containing 10% FBS is added to the cortex to stop the reaction. The cortex is then centrifuged at 2500 rpm for 2 minutes. The supernatant is removed and 2 ml of Neural Basal Media (NBM) containing 2% B27 (vol/vol) and 0.25% Glutamax (vol/vol) is added to the cortex which is then re-suspended by pipetting up and down. Next, the cortex is triturated with previously fire polished glass pipettes, each with a successive smaller opening. After triturating, the cortex is once again centrifuged at 2500 rpm for 2 minutes. The supernatant is then removed and the cortex pellet re-suspended with 2 ml of NBM containing B27 and Glutamax. The cell suspension is then passed through a 40 um nylon cell strainer. Next the cells are counted. The neurons are now ready for loading into the neuron microfluidic device.

In this video we demonstrate the preparation of E18 Cortical Rat Neurons.

Microfluidic 장치에서 Compartmentalization에 대한 E18 두피 랫 뉴런 준비

In this video, we demonstrate the preparation of E18 cortical rat neurons. E18 cortical rat neurons are obtained from E18 fetal rat cortex previously ...

Cell Capture Using a Microfluidic Device

Microfluidic 장치를 사용하여 셀 캡처

Assessing Neural Stem Cell Motility Using an Agarose Gel-based Microfluidic Device

While microfluidic technology is reaching a new level of maturity for macromolecular assays, cell-based assays are still at an infant stage1. This is largely due to the difficulty with which one can create a cell-compatible and steady microenvironment using conventional microfabrication techniques and materials. We address this problem via the introduction of a novel microfabrication material, agarose gel, as the base material for the microfluidic device. Agarose gel is highly malleable, and permeable to gas and nutrients necessary for cell survival, and thus an ideal material for cell-based assays. We have shown previously that agarose gel based devices have been successful in studying bacterial and neutrophil cell migration2. In this report, three parallel microfluidic channels are patterned in an agarose gel membrane of about 1mm thickness. Constant flows with media/buffer are maintained in the two side channels using a peristaltic pump. Cells are maintained in the center channel for observation. Since the nutrients and chemicals in the side channels are constantly diffusing from the side to center channel, the chemical environment of the center channel is easily controlled via the flow along the side channels. Using this device, we demonstrate that the movement of neural stem cells can be monitored optically with ease under various chemical conditions, and the experimental results show that the over expression of epidermal growth factor receptors (EGFR) enhances the motility of neural stem cells. Motility of neural stem cells is an important biomarker for assessing cells aggressiveness, thus tumorigenic factor3. Deciphering the mechanism underlying NSC motility will yield insight into both disorders of neural development and into brain cancer stem cell invasion.

We demonstrate that the over expression of epidermal growth factor receptors (EGFR) enhances the motility of neural stem cells(NSCs) using a novel agarose gel based microfluidic device. This technology can be readily adaptable to other mammalian cell systems where cell sources are scarce, such as human neural stem cells, and the turn around time is critical.

아가로 오스 겔 기반 Microfluidic 장치를 사용 신경 줄기 세포의 운동성 평가

While microfluidic technology is reaching a new level of maturity for macromolecular assays, cell-based assays are still at an infant stage1. This is ...

A Multi-compartment CNS Neuron-glia Co-culture Microfluidic Platform

We present a novel multi-compartment neuron co-culture microsystem platform for in vitro CNS axon-glia interaction research, capable of conducting up to six independent experiments in parallel for higher-throughput. We developed a new fabrication method to create microfluidic devices having both micro and macro scale structures within the same device through a single soft-lithography process, enabling mass fabrication with good repeatability.
The multi-compartment microfluidic co-culture platform is composed of one soma compartment for neurons and six axon/glia compartments for oligodendrocytes (OLs). The soma compartment and axon/glia compartments are connected by arrays of axon-guiding microchannels that function as physical barriers to confine neuronal soma in the soma compartment, while allowing axons to grow into axon/glia compartments. OLs loaded into axon/glia compartments can interact only with axons but not with neuronal soma or dendrites, enabling localized axon-glia interaction studies. The microchannels also enabled fluidic isolation between compartments, allowing six independent experiments to be conducted on a single device for higher throughput.
Soft-lithography using poly(dimethylsiloxane) (PDMS) is a commonly used technique in biomedical microdevices. Reservoirs on these devices are commonly defined by manual punching. Although simple, poor alignment and time consuming nature of the process makes this process not suitable when large numbers of reservoirs have to be repeatedly created. The newly developed method did not require manual punching of reservoirs, overcoming such limitations. First, seven reservoirs (depth: 3.5 mm) were made on a poly(methyl methacrylate) (PMMA) block using a micro-milling machine. Then, arrays of ridge microstructures, fabricated on a glass substrate, were hot-embossed against the PMMA block to define microchannels that connect the soma and axon/glia compartments. This process resulted in macro-scale reservoirs (3.5 mm) and micro-scale channels (2.5 &mu;m) to coincide within a single PMMA master. A PDMS replica that served as a mold master was obtained using soft-lithography and the final PDMS device was replicated from this master.
Primary neurons from E16-18 rats were loaded to the soma compartment and cultured for two weeks. After one week of cell culture, axons crossed microchannels and formed axonal only network layer inside axon/glia compartments. Axons grew uniformly throughout six axon/glia compartments and OLs from P1-2 rats were added to axon/glia compartments at 14 days in vitro for co-culture.

We developed a novel multi-compartment neuron co-culture microsystem platform for in vitro CNS axon-glia interaction research. The platform is capable of conducting up to six independent experiments in parallel and was fabricated using a newly developed macro/micro hybrid fabrication method.

다중 구획 CNS 뉴런 - glia 공동 문화 Microfluidic 플랫폼

We present a novel multi-compartment neuron co-culture microsystem platform for in vitro CNS axon-glia interaction research, capable of conducting up to ...

Microfluidic Co-culture of Epithelial Cells and Bacteria for Investigating Soluble Signal-mediated Interactions

The human gastrointestinal (GI) tract is a unique environment in which intestinal epithelial cells and non-pathogenic (commensal) bacteria coexist. It has been proposed that the microenvironment that the pathogen encounters in the commensal layer is important in determining the extent of colonization. Current culture methods for investigating pathogen colonization are not well suited for investigating this hypothesis as they do not enable co-culture of bacteria and epithelial cells in a manner that mimics the GI tract microenvironment. Here we describe a microfluidic co-culture model that enables independent culture of eukaryotic cells and bacteria, and testing the effect of the commensal microenvironment on pathogen colonization. The co-culture model is demonstrated by developing a commensal Escherichia coli biofilm among HeLa cells, followed by introduction of enterohemorrhagic E. coli (EHEC) into the commensal island, in a sequence that mimics the sequence of events in GI tract infection.

This protocol describes a microfluidic co-culture model for simultaneous and localized culture of epithelial cells and bacteria. This model can be used for investigating the role of different soluble molecular signals on pathogenesis as well as screen the effectiveness of putative probiotic bacterial strains.

Microfluidic 가용성식이 중재 신호의 상호 작용을 조사하는 상피 세포 박테리아의 공동 문화

The human gastrointestinal (GI) tract is a unique environment in which intestinal epithelial cells and non-pathogenic (commensal) bacteria coexist. It has ...

A Microfluidic Device for Quantifying Bacterial Chemotaxis in Stable Concentration Gradients

Chemotaxis allows bacteria to approach sources of attractant chemicals or to avoid sources of repellent chemicals. Bacteria constantly monitor the concentration of specific chemoeffectors by comparing the current concentration to the concentration detected a few seconds earlier. This comparison determines the net direction of movement. Although multiple, competing gradients often coexist in nature, conventional approaches for investigating bacterial chemotaxis are suboptimal for quantifying migration in response to concentration gradients of attractants and repellents. Here, we describe the development of a microfluidic chemotaxis model for presenting precise and stable concentration gradients of chemoeffectors to bacteria and quantitatively investigating their response to the applied gradient. The device is versatile in that concentration gradients of any desired absolute concentration and gradient strength can be easily generated by diffusive mixing. The device is demonstrated using the response of Escherichia coli RP437 to gradients of amino acids and nickel ions.

This protocol describes the development of a microfluidic device for investigating bacterial chemotaxis in stable concentration gradients of chemoeffectors.