Name: capillary-based centrifugal microfluidic device for size-controllable formation of monodisperse microdroplets
Uploaded: 2016-02-22T15:00:00.000+00:00
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Description: Watch this Scientific Journal Video about Capillary-based Centrifugal Microfluidic Device for Size-controllable Formation of Monodisperse Microdroplets at JoVE.com

This work was supported by the PRESTO "Design and Control of Cellular Functions" research area of the Japan Science and Technology Agency (JST), a Grant-in-Aid for Scientific Research of Innovative Areas "Molecular Robotics" (Project No. 24104002) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan, Grant-in-Aid for Young Scientists (A) (Project No. 24680033) and Scientific Research (B) (Project No. 26280097) from the Japan Society for the Promotion of Science (JSPS), and the Creative Design for Bioscience and Biotechnology course of the School of Bioscience and Biotechnology at Tokyo Tech.

모세관 기반 미세 유체 장치 1. 제작 <ol><li> 홀더의 설정 참고 : 홀더 디자인 그림 2A로 표시됩니다. <ol><li> 홀더의 네 각 디스크 잘라 (도 2A (Ⅰ) - (IV)) 밀링 머신을 사용하여 2 mm 두께의 플레이트를 폴리 아세탈 수지. (SH) 직경은 1.8 mm 고정 홈 (I)의 디스크 (1)의 직경 8.5 mm, 모세관 구멍 (CH)의 직경 1.3 mm; 홀더 네 디스크 각각에 대해 다음의 기준을 사용 (ⅱ)이 디스크 직경 8.7 mm, CH 직경 2.0 mm, SH 직경 1.8 mm; (ⅲ) 디스크 (3)의 직경 8.7 mm, CH 직경 0.5 mm, SH 직경 1.8 mm; 및 (ⅳ) 4 장 직경 9.1 mm, 지름 1.0 mm CH, SH 직경 1.8 mm. </li><li> M2 (40) 나사 (그림 2B)를 × 사용하여 홀더를 조립합니다. 홀더 (도 2B)의 저부는도 2a (i), (ii) 및 상부 (도 디스크 (1)과 디스크 (2)로 구성홀더 URE의 2B)를 디스크 (3)와도 2a에 4 장 구성 (III), (IV). <ol><li> 홀더의 하부를 구성하는 각 디스크 (1)의 세 SH에 나사를 삽입 2. 쓰레드 부의 부분을 협지하여 나사를 짧게. 0.9 cm (상기 홀더의 바닥 부와 동일한 길이)에 나사의 길이를 유지한다. </li><li> 홀더의 상부를 구성하는 각각의 디스크의 3 개의 SH에 나사를 삽입 4. 나사부의 일부를 협지하여 나사를 짧게. 0.7 cm (홀더의 상부와 같은 길이)에 나사의 길이를 유지한다. </li><li> 홀더 조립 긴 나사를 사용하여 홀더의 하부 및 상부에 참여. 참고 : 정확한 홀더의 각 부분의 길이를 유지 : 하단 부분은 0.9 cm입니다; 상부는 0.7 cm (그림 2B)입니다. </li></ol></li></ol></li><li> 유리 모세관의 제작 <ol><li> 내부 유리 모세관 (외경 (OD) / 내경 (ID) : 1.0 / 0.6 mm), 및 외부 유리 모세관 (OD / ID : 2.0 / 1.12 mm) 두 개의 유리 모세관의 형식을 사용합니다. </li><li> 3 등분으로 외부 유리 모세관을 분할하는 유리 절단기를 사용하여 두 개의 동일한 조각으로 내부 유리 모세관을 분할하는 유리 커터를 사용합니다. </li><li> 유리 모세관 풀러 (도 3a)을 사용하여 각각의 분할 된 내측 및 외측 유리 모세관을 선명. 최대의 풀러의 무게를 설정합니다. 외부 유리 모세관 60도, 내부 모세관 70도에서 풀러의 열 레벨을 설정합니다. 조심스럽게 유리 모세관을 선명하게. <ol><li> 유리 모세관의 잘록한 부분에서 끝의 길이를 유지 : 내부 모세관은 1.5-1.8 cm이다 외부 모세관은 0.8-1.0 cm (그림 3C)입니다. 이 길이는 기재의 길이보다 짧거나 길면, 풀러 열 조정하여주십시오. </li></ol></li><li> 에프IX microforge에 내부 또는 외부 유리 모세관 테이프 (그림 3B)를 사용하여 서있다. </li><li> 세 단계 (도 3b)에 microforge를 사용하여 유리 모세관의 끝 부분을 잘라 (ⅰ) 백금선에 유리 비드를 유리 모세관의 끝을 터치 (II) 발 밟아 백금선 가열 1-2 초 동안 스위치, 및 (iii) 1 ~ 2 초 후에, 백금선을 냉각시켜 유리 모세관의 끝 부분을 잘라. <ol><li> 각각 내부 (d 개의 i) 및 외부 (D 오) 모세관 구멍의 직경을 조정합니다. 내측 유리 모세관의 오리피스 직경, 10 5, 20 ㎛의 (d에서 I = 5, 10, 20 μm의)과 외부 유리 모세관의 (d 오)이 실험에서 60 μm의 (라 오 = 60 μm의)입니다. 참고 : 유리 모세관은 일회용이다. 유리 capillar의 제조를 반복하여이거 야. </li></ol></li></ol></li></ol> 생성 W / O의 미세 방울 2. 절차 <ol><li> 오일 계면 활성제와 외부 유리 모세관을 입력합니다. 오일 및 계면 활성제의 혼합물은 헥사 (w / w)이 실험 (도 4a)의 소르 비탄 모노 올레 에이트 2 %를 함유한다. 참고 : 오일과 계면 활성제의 많은 조합이있다 (예를 들어, 오일 플루오르 또는 탄산 일 수있다 계면 활성제는 이온 성 비이 온성, 또는 플루오로 할 수있다). <ol><li> 외부 유리 모세관에 헥사 데칸 포함 소르 비탄 모노 올레 에이트의 10 μl를 소개합니다. 도 4a에서, 외부 유리 모세관 (d 오)의 오리피스 직경은 60 μm의 (d 오 = 60 μm의)이다. 유리 모세관의 구멍을 조정하려면, 단계 1.2.4-1.2.5로 돌아갑니다. </li></ol></li><li> 홀더 (그림 4B)의 바닥 부분에 외부 모세관을 설정합니다. </li><li> 드라모세관 작용에 의해 내 유리 모세관 (도 4C)으로 수용액 약 0.1 μL w. 도 4c에서, 내부 유리 모세관 (d 개의 I)의 구멍 직경은 10 μm의가 (d는 내가 10 μm의 =)입니다. 유리 모세관의 구멍을 조정하려면, 단계 1.2.4-1.2.5로 돌아갑니다. </li><li> 홀더 (도 4D-a)의 상단부에 내측 모세관을 설정한다. 외부 모세관 (그림 4D-)에 내부 모세관을 삽입합니다. 그림에서와 같이 흰색 도트 원을 보면 4D-는, 외부 모세관 내부의 내부 모세관의 위치 (외부 모세관 (μm의 w) = (130)의 내부 직경) 관찰 (그림 4D-B를 C) 디지털 현미경을 사용하여 . 외측 모세관의 내부 모세관의 위치는 w = 100 ~ 150 ㎛, 설정해야한다. 참고 : 내부의 위치를 변경하려면외부 모세관의 모세관는 홀더의 상부에 나사를 켜십시오. 이것에 의해, 거리 (W)을 정밀하게 제어 할 수있다. </li><li> 1.5 ml의 샘플을 마이크로 튜브의 바닥에 헥사 함유 소르 비탄 모노 올 레이트 (w / w 2 %) 100 μl를 소개한다. 샘플의 마이크로 튜브 (도 4E-a)에서, 내측 및 외측 모세관으로 홀더를 설치한다. 공기 - 오일 인터페이스 (그림 4E-B)에서 멀리 유지하기 위해 외부 모세관을 확인하십시오. </li><li> 원심 분리기 (그림 4 층) 미세 방울을 생성하기 위해 1 ~ 2 초 동안 1,600 XG의 중력에 탁상 스윙 아웃 형 미니 원심 분리기를 사용하여 샘플 마이크로 튜브. 실온에서 모든 실험을 수행한다. 참고 : 스윙 아웃 형 원심 분리기를 사용합니다. 액적 샘플 마이크로 튜브의 측벽에 충돌하고, 고정 각도 타입의 원심 분리를 사용할 때 분해된다. </li><li> 천천히 피펫 W / O 방울을 작성하고, 유리 (S)에 올려LIDE. </li><li> 미세 방울의 캡처 된 이미지가 디지털 현미경 (배율 200 배)을 이용하여 생성. </li></ol>

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No conflicts of interest are declared.

이 장치를 사용하면, / O W 미세 방울은 단 분산 제트 흐름 (17)의 플래 토우 레일리 불안정성에 의해 발생되었다. 현미경 검사 위성 방울의 존재를 공개하지 않았다. 장치의 제조에있어서, 세 가지 중요한 단계가 성공적으로 단 분산 W / O의 미세 방울을 생성하는 것이 필수적이다. 우선, 오일 및 계면 활성제의 수용액의 직선 유동을 공급하는 네 개의 디스크 모세관 구멍이 동심원 형태로 배?...

W / O 미세 방울 1-5 단백질 합성 (6), 단백질 결정화 (7), 에멀젼 PCR 8, 9, 세포 캡슐화 (10), 인공 세포와 같은 시스템 5,6의 건설을 포함한 생화학 및 생명 공학의 연구에 많은 중요한 응용 프로그램을 가지고있다. 이러한 응용을위한 W / O의 미세 방울을 생산 중요한 기준은 사이즈 W / O의 미세 방울의 단순 분산도를 제어한다. 미세 유체 장치의 제조 단 분산의 크기가 제어 W / 미세 방울 (11)은 공동 (12, 13)에있어서 흐르는 흐름 포커싱 방법 14,15 및 마이크로에 T 접합 방법 (16)에 기초 이여. 이러한 방법은 매우 단 분산 W / O의 미세 방울을 생성하지만, 미세 처리가 마이크로 채널 제조 복잡한 처리 및 전문 기술을 필요로하고, 또한 시료 용액을 다량 (최소 수백이 필요81; 때문에 마이크로에 시료 용액을 실시 주사기 펌프와 튜브 불가피 데드 볼륨 l). / O 미세 방울가 필요하다 (W) 따라서, 사용하기 쉬운과 낮은 죽은 볼륨 방법은 단 분산 생성합니다. 이 논문은, 실험 절차의 동영상과 함께 / O 미세 방울 (그림 1) W 단 분산 셀 크기의 생성 원심 모세관 기반의 축 대칭 공동 흐르는 미세 유체 장치 (17)에 대해 설명합니다. 이 간단한 방법은 크기의 단 분산 및 크기 제어를 달성 할 수있다. 그냥 탁상 미니 원심 분리기 및 샘플링 마이크로 튜브에 고정 된 모세관 기반의 축 대칭 공동 흐르는 미세 유체 장치가 필요합니다. 우리의 방법은 단지 매우 소량 (0.1 μL)를 필요로하고, 샘플의 상당한 양을 낭비하지 않는다.

<table><tbody><tr><td>2-mm-thick polyacetal plastic plate</td><td>Tool</td><td>Nikkyo Technos, Co., Ltd. (Japan)</td><td>244-6432-08</td></tr><tr><td>Milling machine</td><td>Tool</td><td>Roland DG Co., Ltd. (Japan)</td><td>MDX-40A</td></tr><tr><td>End Mill RSE230-0.5*2.5</td><td>Tool</td><td>NS Tool Co., Ltd. (Japan)</td><td>01-00644-00501</td></tr><tr><td>M2*40 screws</td><td>Tool</td><td>Jujo Synthetic Chemistry Labo. (Japan)</td><td>0001-024</td></tr><tr><td>Glass Capillry Puller</td><td>Tool</td><td>Narishige (Japan)</td><td>PC-10</td></tr><tr><td>Microforge</td><td>Tool</td><td>Narishige (Japan)</td><td>MF-900</td></tr><tr><td>Inner Glass Capillary</td><td>Tool</td><td>Narishige (Japan)</td><td>G-1</td></tr><tr><td>Outer Glass Capillary</td><td>Tool</td><td>World Precision Instruments Inc. (USA)</td><td>1B200-6</td></tr><tr><td>1.5 ml Sample tube</td><td>Tool</td><td>INA OPTIKA CO.,LTD (Japan)</td><td>ST-0150F</td></tr><tr><td>Hexadecane</td><td>Reagent</td><td>Wako Pure Chemical Industries Ltd. (Japan)</td><td>080-03685&amp;nbsp;</td></tr><tr><td>Sorbitan monooleate (Span 80)</td><td>Reagent</td><td>Tokyo Chemical Industry Co., Ltd. (Japan)</td><td>S0060</td></tr><tr><td>Milli Q system</td><td>Reagent</td><td>Merck Millipore Corporation (Germany)</td><td>ZRQSVP030</td></tr><tr><td>Swinging-out-type Mini-centrifuge</td><td>Tool</td><td>Hitech Co., Ltd. (Japan)</td><td>ATT101</td></tr><tr><td>Digital Microscope</td><td>Tool</td><td>KEYENCE Corporation (Japan)</td><td>VHX-2001</td></tr></tbody></table>

capillary-based centrifugal microfluidic device for size-controllable formation of monodisperse microdroplets

여기서는 모세관 기반 원심 미세 유동 장치를 이용하여 제어 할 수있는 크기, 단 분산, W / 출력 미세 방울의 신속한 제조를위한 간단한 방법을 보여준다. W / O의 미세 방울은 최근 화학 실험 소형화 에이블 강력한 방법에 사용되어왔다. / O 미세 방울 필요 W 따라서, 다목적 현상 방법은 단 분산 얻었다. 우리는 모세관 기반 원심 선대칭 공동 흐르는 미세 유동 장치에 기초하여 / O의 미세 방울 W 단 분산 생성하기위한 방법을 개발 하였다. 우리는 모세관 구멍을 조절함으로써 미세 방울의 크기를 제어하는​​데 성공했다. 우리의 방법은, 밀봉 용 시료 액의 소량 (0.1 μL)를 필요로 사용하기 쉬운 다른 마이크로 유체 기술을보다 장비를 필요로 초당 W / O의 미세 방울 수백 수천의 수의 생산을 가능하게 . 우리는이 방법이 소중한 생물들 필요 생물학적 연구를 도움이 될 것 기대신속한 정량 분석​​ 생화학 및 생물학적 연구를위한 샘플의 부피를 보존하여 amples.

본 연구에서는 모세관 기반 원심 미세 유동 장치 (도 1)를 사용하여 셀 크기 W / O의 미세 방울의 생성을위한 간단한 방법을 제시한다. 미세 유동 장치가 모세관 홀더 (도 2B)로 구성하고, 두 유리 모세관 (도 3c에 내부 및 외부 유리 모세관), 및 계면 활성제를 포함하는 기름을 포함하는 마이크로 튜브. 우리는 내측 유리 모세관에 시료 용?...

Watch this Scientific Journal Video about Capillary-based Centrifugal Microfluidic Device for Size-controllable Formation of Monodisperse Microdroplets at JoVE.com

Capillary-based Centrifugal Microfluidic Device for Size-controllable Formation of Monodisperse Microdroplets

Here, we demonstrate a simple production method for size-controllable, monodisperse, water-in-oil (W/O) microdroplets using a capillary-based centrifugal microfluidic device. This method requires only a small sample volume and enables high-yield production. We expect this method will be useful for rapid biochemical and cellular analyses.

단 분산 미세 방울의 크기가 제어 형성을위한 모세관 기반 원심 미세 유체 장치

Here, we demonstrate a simple method for the rapid production of size-controllable, monodisperse, W/O microdroplets using a capillary-based centrifugal ...

capillary-based-centrifugal-microfluidic-device-for-size-controllable

Research

JoVE Journal

Engineering

10.3K 조회수.  Tokyo Institute of Technology. Here, we demonstrate a simple production method for size-controllable, monodisperse, water-in-oil (W/O) microdroplets using a capillary-based centrifugal microfluidic device. This method requires only a small sample volume and enables high-yield production. We expect this method will be useful for rapid biochemical and cellular analyses.

비디오: Capillary-based Centrifugal Microfluidic Device for Size-controllable Formation of Monodisperse Microdroplets

Fluorescence detection methods for microfluidic droplet platforms

The development of microfluidic platforms for performing chemistry and biology has in large part been driven by a range of potential benefits that accompany system miniaturisation. Advantages include the ability to efficiently process nano- to femoto- liter volumes of sample, facile integration of functional components, an intrinsic predisposition towards large-scale multiplexing, enhanced analytical throughput, improved control and reduced instrumental footprints.1
In recent years much interest has focussed on the development of droplet-based (or segmented flow) microfluidic systems and their potential as platforms in high-throughput experimentation.2-4 Here water-in-oil emulsions are made to spontaneously form in microfluidic channels as a result of capillary instabilities between the two immiscible phases. Importantly, microdroplets of precisely defined volumes and compositions can be generated at frequencies of several kHz. Furthermore, by encapsulating reagents of interest within isolated compartments separated by a continuous immiscible phase, both sample cross-talk and dispersion (diffusion- and Taylor-based) can be eliminated, which leads to minimal cross-contamination and the ability to time analytical processes with great accuracy. Additionally, since there is no contact between the contents of the droplets and the channel walls (which are wetted by the continuous phase) absorption and loss of reagents on the channel walls is prevented.
Once droplets of this kind have been generated and processed, it is necessary to extract the required analytical information. In this respect the detection method of choice should be rapid, provide high-sensitivity and low limits of detection, be applicable to a range of molecular species, be non-destructive and be able to be integrated with microfluidic devices in a facile manner. To address this need we have developed a suite of experimental tools and protocols that enable the extraction of large amounts of photophysical information from small-volume environments, and are applicable to the analysis of a wide range of physical, chemical and biological parameters. Herein two examples of these methods are presented and applied to the detection of single cells and the mapping of mixing processes inside picoliter-volume droplets. We report the entire experimental process including microfluidic chip fabrication, the optical setup and the process of droplet generation and detection.

Droplet-based microfluidic platforms are promising candidates for high throughput experimentation since they are able to generate picoliter, self-compartmentalized vessels inexpensively at kHz rates. Through integration with fast, sensitive and high resolution fluorescence spectroscopic methods, the large amounts of information generated within these systems can be efficiently extracted, harnessed and utilized.

microfluidic 비말 플랫폼 형광 검출 방법

The development of microfluidic platforms for performing chemistry and biology has in large part been driven by a range of potential benefits that ...

연구

생체공학

Fabricating High-viscosity Droplets using Microfluidic Capillary Device with Phase-inversion Co-flow Structure

The generation of monodisperse droplets with high viscosity has always been a challenge in droplet microfluidics. Here, we demonstrate a phase-inversion co-flow device to generate uniform high-viscosity droplets in a low-viscosity fluid. The microfluidic capillary device has a common co-flow structure with its exit connecting to a wider tube. Elongated droplets of the low-viscosity fluid are first encapsulated by the high-viscosity fluid in the co-flow structure. As the elongated low-viscosity droplets flow through the exit, which is treated to be wetted by the low-viscosity fluid, phase inversion is then induced by the adhesion of the low viscosity droplets to the tip of the exit, which results in the subsequent inverse encapsulation of the high-viscosity fluid. The size of the resultant high-viscosity droplets can be adjusted by changing the flow rate ratio of the low-viscosity fluid to the high-viscosity fluid. We demonstrate several typical examples of the generation of high-viscosity droplets with a viscosity up to 11.9 Pas, such as glycerol, honey, starch, and polymer solution. The method provides a simple and straightforward approach to generate monodisperse high-viscosity droplets, which may be used in a variety of droplet-based applications, such as materials synthesis, drug delivery, cell assay, bioengineering, and food engineering.

A phase-inversion co-flow device is demonstrated to generate monodisperse high-viscosity droplets above 1 Pas, which is difficult to realize in droplet microfluidics.

위상 반전 공동 흐름 구조와 미세 모 세관 장치를 사용 하 여 조작 높은 점도 방울

The generation of monodisperse droplets with high viscosity has always been a challenge in droplet microfluidics. Here, we demonstrate a phase-inversion ...

화학

Generation of Size-controlled Poly (ethylene Glycol) Diacrylate Droplets via Semi-3-Dimensional Flow Focusing Microfluidic Devices

Uniform and size-controllable poly (ethylene glycol) diacrylate (PEGDA) droplets could be produced via the flow focusing process in a microfluidic device. This paper proposes a semi-three-dimensional (semi-3D) flow-focusing microfluidic chip for droplet formation. The polydimethylsiloxane (PDMS) chip was fabricated using the multi-layer soft lithography method. Hexadecane containing surfactant was used as the continuous phase, and PEGDA with the ultraviolet (UV) photo-initiator was the dispersed phase. Surfactants allowed the local surface tension to drop and formed a more cusped tip which promoted breaking into tiny micro-droplets. As the pressure of dispersed phase was constant, the size of droplets became smaller with increasing continuous phase pressure before dispersed phase flow was broken off. As a result, droplets with size variation from 1 &#181;m to 80 &#181;m in diameter could be selectively achieved by changing the pressure ratio in two inlet channels, and the average coefficient of variation was estimated to be below 7%. Furthermore, droplets could turn into micro-beads by UV exposure for photo-polymerization. Conjugating biomolecules on such micro-beads surface have many potential applications in the fields of biology and chemistry.

Generation of Size-controlled Poly ethylene Glycol Diacrylate Droplets via Semi-3-Dimensional Flow Focusing Microfluidic Devices

Here, we present a protocol to illustrate the fabrication processes and verifying experiments of a semi-three-dimensional (semi-3D) flow-focusing microfluidic chip for droplet formation.

크기 제어 폴 리 (에틸렌 글리콜) 빛 방울 장치를 통해 세미 3 차원 흐름 초점 미세의 세대

Uniform and size-controllable poly (ethylene glycol) diacrylate (PEGDA) droplets could be produced via the flow focusing process in a microfluidic device. ...

공학

A Microfluidic Chip for ICPMS Sample Introduction

This protocol discusses the fabrication and usage of a disposable low cost microfluidic chip as sample introduction system for inductively coupled plasma mass spectrometry (ICPMS). The chip produces monodisperse aqueous sample droplets in perfluorohexane (PFH). Size and frequency of the aqueous droplets can be varied in the range of 40 to 60 &#181;m and from 90 to 7,000&#160;Hz, respectively. The droplets are ejected from the chip with a second flow of PFH and remain intact during the ejection. A custom-built desolvation system removes the PFH and transports the droplets into the ICPMS. Here, very stable signals with a narrow intensity distribution can be measured, showing the monodispersity of the droplets. We show that the introduction system can be used to quantitatively determine iron in single bovine red blood cells. In the future, the capabilities of the introduction device can easily be extended by the integration of additional microfluidic modules.

We present a discrete droplet sample introduction system for inductively coupled plasma mass spectrometry (ICPMS). It is based on a cheap and disposable microfluidic chip that generates highly monodisperse droplets in a size range of 40&#8722;60 &#181;m at frequencies from 90 to 7,000 Hz.

ICPMS 샘플 소개를위한 미세 유체 칩

This protocol discusses the fabrication and usage of a disposable low cost microfluidic chip as sample introduction system for inductively coupled plasma ...

High Speed Droplet-based Delivery System for Passive Pumping in Microfluidic Devices

A novel microfluidic system has been developed that uses the phenomenon of passive pumping along with a user controlled droplet based fluid delivery system. Passive pumping is the phenomenon by which surface tension induced pressure differences drive fluid movement in closed channels. The automated fluid delivery system consists of a set of voltage controlled valves with micro-nozzles connected to a fluid reservoir and a control system. These voltage controlled valves offer a volumetrically precise way to deliver fluid droplets to the inlet of a microfluidic device in a high frequency manner. Based on the  dimensions demonstrated in the current study example, the system is capable of flowing 4 milliliters per minute (through a 2.2mm by 260um cross-sectional channel). Based on these same channel dimensions, fluid exchange of a point inside the channel can be achieved in as little as eight milliseconds. It is observed that there is interplay between momentum of the system (imparted by a combination of the droplets created by the valves and the fluid velocity in the channel), and the surface tension of the liquid. Where momentum provides velocity to the fluid flow (or vice-versa), equilibration of surface tension at the inlet provides a sudden stop to any flow. This sudden stop allows the user to control the flow characteristics of the channel and opens the door for a variety of biological applications, ranging anywhere from reagent delivery to drug-cell studies. It is also observed that when nozzles are aimed at the inlet at shallow angles, the droplet momentum can cause additional interesting fluid phenomena, such as mixing of multiple droplets in the inlet.

A novel microfluidic system has been developed using the phenomenon of passive pumping and a user controlled fluid delivery system. This microfluidic system has the potential to be used in a wide variety of biological applications given its low cost, ease of use, volumetric precision, high speed, repeatability and automation.

Microfluidic 장치에 수동 펌핑을위한 고속 비말 기반 전달 시스템

A novel microfluidic system has been developed that uses the phenomenon of passive pumping along with a user controlled droplet based fluid delivery ...

생물학

A Microfluidic Platform for Precision Small-volume Sample Processing and Its Use to Size Separate Biological Particles with an Acoustic Microdevice

A major advantage of microfluidic devices is the ability to manipulate small sample volumes, thus reducing reagent waste and preserving precious sample. However, to achieve robust sample manipulation it is necessary to address device integration with the macroscale environment. To realize repeatable, sensitive particle separation with microfluidic devices, this protocol presents a complete automated and integrated microfluidic platform that enables precise processing of 0.15&#8211;1.5 ml samples using microfluidic devices. Important aspects of this system include modular device layout and robust fixtures resulting in reliable and flexible world to chip connections, and fully-automated fluid handling which accomplishes closed-loop sample collection, system cleaning and priming steps to ensure repeatable operation. Different microfluidic devices can be used interchangeably with this architecture. Here we incorporate an acoustofluidic device, detail its characterization, performance optimization, and demonstrate its use for size-separation of biological samples. By using real-time feedback during separation experiments, sample collection is optimized to conserve and concentrate sample. Although requiring the integration of multiple pieces of equipment, advantages of this architecture include the ability to process unknown samples with no additional system optimization, ease of device replacement, and precise, robust sample processing.

This protocol describes a system architecture for performing automated small volume (0.15&#8211;1.5 ml) particle separations using a microfluidic device, and discusses methods to optimize acoustofluidic device performance and operation.

미세 유체 정밀 작은 볼륨 샘플 처리를위한 플랫폼 및 음향 마이크로 디바이스로 구분 생물 입자 크기의 사용

A major advantage of microfluidic devices is the ability to manipulate small sample volumes, thus reducing reagent waste and preserving precious sample. ...

A Femtoliter Droplet Array for Massively Parallel Protein Synthesis from Single DNA Molecules

Advances in spatial resolution and detection sensitivity of scientific instrumentation make it possible to apply small reactors for biological and chemical research. To meet the demand for high-performance microreactors, we developed a femtoliter droplet array (FemDA) device and exemplified its application in massively parallel cell-free protein synthesis (CFPS) reactions. Over one million uniform droplets were readily generated within a finger-sized area using a two-step oil-sealing protocol. Every droplet was anchored in a femtoliter microchamber composed of a hydrophilic bottom and a hydrophobic sidewall. The hybrid hydrophilic-in-hydrophobic structure and the dedicated sealing oils and surfactants are crucial for stably retaining the femtoliter aqueous solution in the femtoliter space without evaporation loss. The femtoliter configuration and the simple structure of the FemDA device allowed minimal reagent consumption. The uniform dimension of the droplet reactors made large-scale quantitative and time-course measurements convincing and reliable. The FemDA technology correlated the protein yield of the CFPS reaction with the number of DNA molecules in each droplet. We streamlined the procedures about the microfabrication of the device, the formation of the femtoliter droplets, and the acquisition and analysis of the microscopic image data. The detailed protocol with the optimized low running cost makes the FemDA technology accessible to everyone who has standard cleanroom facilities and a conventional fluorescence microscope in their own place.

The overall goal of the protocol is to prepare over one million ordered, uniform, stable, and biocompatible femtoliter droplets on a 1 cm2 planar substrate that can be used for cell-free protein synthesis.

단일 DNA 분자에서 대규모 병렬 단백질 합성을 위한 펨토리터 물방울 배열

Advances in spatial resolution and detection sensitivity of scientific instrumentation make it possible to apply small reactors for biological and ...

Glass-Based Devices to Generate Drops and Emulsions

In this manuscript, three different step-by-step protocols to generate highly monodisperse emulsion drops using glass-based microfluidics are described. The first device is built for the generation of simple drops driven by gravity. The second device is designed to generate emulsion drops in a coflowing scheme. The third device is an extension of the coflowing device with the addition of a third liquid that acts as an electric ground, allowing the formation of electrified drops that subsequently discharge. In this setup, two of the three liquids have an appreciable electrical conductivity. The third liquid mediates between these two and is a dielectric. A&#160;voltage difference applied between the two conducting liquids creates an electric field that couples with hydrodynamic stresses of the coflowing liquids, affecting the jet and drop formation process. The addition of the electric field provides a path to generate smaller drops than in simple coflow devices and for generating particles and fibers with a wide range of sizes.

Here, a protocol to manufacture glass-based microfluidic devices used for generating highly monodisperse emulsions with controlled drop size is presented.

방울과 에멀젼을 생성하는 유리 기반 장치

In this manuscript, three different step-by-step protocols to generate highly monodisperse emulsion drops using glass-based microfluidics are described. ...

Generation of Dynamical Environmental Conditions using a High-Throughput Microfluidic Device

Mimicking in vivo environmental conditions is crucial for in vitro studies on complex life machinery. However, current techniques targeting live cells and organs are either highly expensive, like robotics, or lack nanoliter volume and millisecond time accuracy in liquid manipulation. We herein present the design and fabrication of a microfluidic system, which consists of 1,500 culture units, an array of enhanced peristaltic pumps and an on-site mixing modulus. To demonstrate the capacities of the microfluidic device, neural stem cell (NSC) spheres are maintained in the proposed system. We observed that when the NSC sphere is exposed to CXCL in day 1 and EGF in day 2, the round-shaped conformation is well maintained. Variation in the input order of 6 drugs causes morphological changes to the NSC sphere and the expression level representative marker for NSC stemness (i.e., Hes5 and Dcx). These results indicate that dynamic and complex environmental conditions have great effects on NSC differentiation and self-renewal, and the proposed microfluidic device is a suitable platform for high throughput studies on the complex life machinery.

We present a microfluidic system for high throughput studies on complex life machinery, which consists of 1500 culture units, an array of enhanced peristaltic pumps and an on-site mixing modulus. The microfluidic chip allows for the analysis of the highly complex and dynamic micro-environmental conditions in vivo.

고처리량 미세 유체 장치를 사용하여 동적 환경 조건의 생성

Mimicking in vivo environmental conditions is crucial for in vitro studies on complex life machinery. However, current techniques targeting live cells and ...

조합 플러그 생산을 위한 미세유체 장치 설정

Bilayer Microfluidic Device for Combinatorial Plug Production

Droplet microfluidics is a versatile tool that allows the execution of a large number of reactions in chemically distinct nanoliter compartments. Such systems have been used to encapsulate a variety of biochemical reactions - from incubation of single cells to implementation of PCR reactions, from genomics to chemical synthesis. Coupling the microfluidic channels with regulatory valves allows control over their opening and closing, thereby enabling the rapid production of large-scale combinatorial libraries consisting of a population of droplets with unique compositions. In this paper, protocols for the fabrication and operation of a pressure-driven, PDMS-based bilayer microfluidic device that can be utilized to generate combinatorial libraries of water-in-oil emulsions called plugs are presented. By incorporating software programs and microfluidic hardware, the flow of desired fluids in the device can be controlled and manipulated to generate combinatorial plug libraries and to control the composition and quantity of constituent plug populations. These protocols will expedite the process of generating combinatorial screens, particularly to study drug response in cells from cancer patient biopsies.

저자 스포트라이트: Integrating Computational and Experimental Approaches in Precision Oncology

The fabrication of a polydimethylsiloxane (PDMS)-based bilayer device for the production of combinatorial libraries in water-in-oil emulsions (plugs) is presented here. The necessary hardware and software required to automate plug production are detailed in the protocol, and the production of a quantitative library of fluorescent plugs is also demonstrated.

조합 플러그 생산을 위한 이중층 미세유체 장치

Droplet microfluidics is a versatile tool that allows the execution of a large number of reactions in chemically distinct nanoliter compartments. Such ...