DOCSLIB.ORG

Polymerase Chain Reaction in Microfluidic Devices

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Polymerase Chain Reaction in Microfluidic Devices Featuring work from the BioNano Technology Lab, As featured in: Dr. Christian D. Ahrberg, Prof. Andreas Manz and Prof. Bong Geun Chung, Sogang University, Korea Polymerase chain reaction in microfl uidic devices We highlight recent developments of polymerase chain reaction (PCR) in microfl uidic devices. Time and space domain microfl uidic devices as well as isothermal nucleic acid amplifi cation and digital PCR microfl uidic devices are discussed. See Bong Geun Chung et al., Lab Chip, 2016, 16, 3866. www.rsc.org/loc Registered charity number: 207890 Lab on a Chip View Article Online CRITICAL REVIEW View Journal | View Issue Polymerase chain reaction in microfluidic devices Christian D. Ahrberg,a Andreas Manzbc and Bong Geun Chung*a Cite this: Lab Chip,2016,16,3866 The invention of the polymerase chain reaction (PCR) has caused a revolution in molecular biology, giving access to a method of amplifying deoxyribonucleic acid (DNA) molecules across several orders of magni- tude. Since the first application of PCR in a microfluidic device was developed in 1998, an increasing num- Received 2nd August 2016, ber of researchers have continued the development of microfluidic PCR systems. In this review, we intro- Accepted 6th September 2016 duce recent developments in microfluidic-based space and time domain devices as well as discuss various designs integrated with multiple functions for sample preparation and detection. The development of iso- DOI: 10.1039/c6lc00984k thermal nucleic acid amplification and digital PCR microfluidic devices within the last five years is also www.rsc.org/loc highlighted. Furthermore, we introduce various commercial microfluidic PCR devices. Introduction naturation, annealing, and extension. In the denaturation Creative Commons Attribution 3.0 Unported Licence. step (≈95 °C), double stranded DNA denaturates into two sin- Since polymerase chain reaction (PCR) was invented by Kary gle strands. Afterwards the temperature is decreased for the Mullis over 30 years ago,1 it has become a valuable tool annealing step (≈56 °C). Primers, which are short, comple- for molecular biology as it can exponentially amplify mentary sequences of DNA, can anneal to the single stranded deoxyribonucleic acid (DNA). Several million copies of DNA target DNA. Furthermore, a polymerase attaches to the com- can be generated from a single molecule by only few cycles of plex; the polymerase is often derived from thermophile or- PCR. A PCR cycle typically consists of three distinct steps, de- ganisms like Thermus aquaticus.2 To increase activity of the polymerase, the temperature is raised for the extension step, typically to around 72 °C. The polymerase gains activity, syn- This article is licensed under a a Department of Mechanical Engineering, Sogang University, Seoul, Korea. E-mail: [email protected] thesizing a second complementary strand of DNA from free b Microfluidics group, KIST-Europe, Saarbrücken, Germany nucleotides in solution. Therefore, an exponential increase of c Mechanotronics Department, Universität des Saarlandes, Saarbrücken, Germany double stranded DNA concentration is obtained by repeating Open Access Article. Published on 06 September 2016. Downloaded 10/4/2021 9:58:02 AM. Christian D. Ahrberg obtained AndreasManziscurrentlypro- his Bachelor and Master Degree fessor at the Saarland Univer- in Chemical and Bioengineering sity, and principal investigator from ETH Zürich. After graduat- at KIST Europe, Saarbrücken ing in 2012 he worked at Bayer Germany. He graduated 1986 Technology Services in Leverku- with PhD from Chemistry ETH sen before starting his PhD un- Zurich Switzerland, was group der supervision of Andreas Manz leader at Ciba-Geigy Ltd. at the Mechatronics department (Novartis) Basel Switzerland of Saarland University in cooper- 1988–1995, professor at Impe- ation with KIST Europe, during rial College London UK 1995– whichheworkedonaportable 2004, head of the ISAS (Leibniz- Christian D. Ahrberg real-time PCR device. Since grad- Andreas Manz Institute of Analytical Sciences) uating in 2016, currently he is a and professor at the Technical postdoctoral researcher at the BioNano Technology Lab in Sogang University Dortmund Germany 2003–2008. He was founder and University, Seoul, Korea. chairman of the RSC journal Lab on a Chip 2001–2008, and sci- entific advisor to Caliper Technologies Inc. (now part of Perkin Elmer) 1995–2000. 3866 | Lab Chip,2016,16,3866–3884 This journal is © The Royal Society of Chemistry 2016 View Article Online Lab on a Chip Critical review three temperature steps. The method was extended to reverse fluidic device was demonstrated by Kopp et al.14 A sample transcription PCR.3 Starting with a ribonucleic acid (RNA) was flown through a microfluidic channel, repeatedly passing molecule, a reverse transcription step is performed to obtain the three temperature zones for PCR. A 10 μL sample could DNA complementary to the RNA strand before PCR. A further perform 20 cycles of PCR within 90 seconds. Numerous appli- modification of PCR introduced fluorescent dyes into the re- cations of PCR in microfluidic devices have been pro- action mixture, allowing the real-time monitoring of reaction posed.15,16 Through the low sample and power consumption, progress.4 This extension of the method allowed the determi- these microfluidic devices are especially interesting for point- nation of initial copies of DNA in the reaction mixture of-care applications. The possibility to combine sample prep- through standard curves. The method is referred to as quan- aration, PCR, and detection in the same automated micro- titative real-time PCR (qrtPCR). Today, a number of PCR ap- fluidic device also promises reduction of handling time as – plications (e.g., diagnostics and medicine,5 7 agricultural sci- well as prevention of sample contamination. This review in- ences,8 and food sciences9) can be found. troduces and discusses various designs of PCR in micro- Microfluidic devices offer small thermal mass, low ther- fluidic device developed within the last five years. First, space mal inertia, and rapid heat transfer. They present a number domain PCR microfluidic devices are discussed before mov- of advantages compared to macroscopic equivalents. The ing on to their counterpart time domain PCR microfluidic de- small volumes reduce sample and reagent consumption, vices. Second, isothermal nucleic acid amplification and digi- leading to inexpensive operation of the systems. However, tal PCR in microfluidic devices are discussed (Table 1). volumes in most cases are still large enough for bulk kinetics to apply. Furthermore, different functions can easily be inte- Space domain PCR grated in microfluidic devices.10 Diffusive mass transport and heat conduction scale with the characteristic length of the When the sample is moved through a microchannel featuring system. While in macroscale systems diffusion and heat con- different temperatures along its length, the temperature is dependent on the position in the channel. Thus, this group Creative Commons Attribution 3.0 Unported Licence. duction are slow for efficient transport, the smaller scales of microfluidic devices allow for utilization of these effects. of devices is referred to as space domain PCR device. Sample Furthermore, these effects are easier to control than convec- preparation can easily be integrated in the microfluidic de- tive heat and mass transport used on larger scales instead. vice. The duration of the thermal steps of PCR and the num- This allows for a better control of processes in microfluidic ber of cycles are predefined by the microfluidic design. The devices. A reduction in size also leads to a higher surface to operation of space domain PCR devices requires movement volume ratio. This means that the exchange areas for mass of the sample along the channel. and heat transport are larger, allowing for faster transport. On the other hand, surface and wall effects become more Serpentine channel designs This article is licensed under a important compared to larger devices. As PCR requires many Since the first application of a microfluidic PCR device, a vari- heating and cooling steps, total analysis time can be signifi- ety of devices improving and evolving the initial design have cantly reduced by miniaturization. Examples of PCR conducted been developed. A polyimide device with three integrated in microfluidic chambers are among the first examples of resistive copper heaters has been presented (Fig. 1A).17 The Open Access Article. Published on 06 September 2016. Downloaded 10/4/2021 9:58:02 AM. PCR in microfluidic devices.11,12 Soon after ligase chain reac- microfluidic device was designed for 30 cycles of PCR with a tion, a method closely related to PCR was shown in the same time ratio of 1 : 1 : 2 for denaturation, annealing, and exten- devices.13 In 1998, the first flow through PCR in a micro- sion step. Due to the low cost of the plastic device, it can be disposed after the experiment preventing cross- contamination that could arise through adsorption of DNA Bong Geun Chung is currently onto the channel walls. In another study, a similar design Associate Professor at Depart- featuring 30 cycles of PCR was created in a polycarbonate ment of Mechanical Engineering substrate using a computer numerical controlled (CNC) verti- in Sogang University, Seoul, Ko- cal milling machine.18 The temperatures were created using rea. He received his Bachelor two copper heating blocks for denaturation and a combined and Master degrees from Han- annealing/extension step. Wu et al. reduced the flow resist of yang University, Seoul, Korea these devices by coating a 0.5 mm inner diameter steel capil- and his Ph.D. degree from Uni- lary with Norland Optical Adhesive 61 (NOA61).19 A smooth, versity of California Irvine, USA hydrophobic inner surface was used to reduce the back pres- in 2007. He was an Instructor at sure. While the previous examples experimented with differ- Department of Medicine in Har- ent substrate materials reducing device costs and optimizing vard Medical School, Boston, flow resistance, other researchers optimized the heaters or Bong Geun Chung USA.
Load more

Recommended publications
  • Digital Microfluidic Devices: the Role of the Dielectric Layer
    Vasco Alexandre Violante Rodrigues Licenciatura em Ciências de Engenharia em Micro e Nanotecnologias Digital Microfluidic devices: the role of the dielectric layer Dissertação para obtenção do Grau de Mestre em Engenharia de Micro e Nanotecnologias Orientador: Professor Doutor Rui Igreja, Dep. Ciência dos Materiais, FCT-UNL Co -orientador: Professor Doutor Hugo Águas, Dep. Ciência dos Materiais, FCT-UNL Júri: Presidente: Prof. Doutor Rodrigo Ferrão de Paiva Martins Arguente: Doutora Rita Maria Mourão Salazar Branquinho Vogal: Prof. Doutor Rui Alberto Garção Barreira do Nascimento Igreja Novembro, 2014 DIGITAL MICROFLUIDICS: THE ROLE OF THE DIELECTRIC LAYER © Vasco Alexandre Violante Rodrigues Faculdade de Ciências e Tecnologia Universidade Nova de Lisboa A Faculdade de Ciências e Tecnologia e a Universidade Nova de Lisboa têm o direito, perpétuo e sem limites geográficos, de arquivar e publicar esta dissertação através de exemplares impressos reproduzidos em papel ou de forma digital, ou por qualquer outro meio conhecido ou que venha a ser inventado, e de a divulgar através de repositórios científicos e de admitir a sua cópia e distribuição com objectivos educacionais ou de investigação, não comerciais, desde que seja dado crédito ao autor e editor. AGRADECIMENTOS É com enorme satisfação que vejo cumprida mais uma etapa da minha vida, tendo sido esta porventura uma das mais exigentes até ao momento. A conclusão deste trabalho encerra em si um capítulo que começou por ser caracterizado por grande incerteza, tendo eu pertencido ao reduzido grupo de pessoas que ingressou pela primeira vez no Mestrado Integrado em Engenharia de Micro e Nanotecnologias, e é com alegria que pertenço ao grupo ainda mais restrito de pessoas que o conclui agora, cinco anos volvidos.
    [Show full text]
  • Recent Advances in Electrowetting Microdroplet Technologies
    Chapter 4 Recent Advances in Electrowetting Microdroplet Technologies Robert W. Barber and David R. Emerson 4.1 Introduction The ability to handle small volumes of liquid, typically from a few picolitres (pL) to a few microlitres (mL), has the potential to improve the performance of complex chemical and biological assays [1–6]. The benefits of miniaturisation are well documented [7–9] and include smaller sample requirements, reduced reagent con- sumption, decreased analysis time, lower power consumption, lower costs per assay and higher levels of throughput and automation. In addition, miniaturisation may offer enhanced functionality that cannot be achieved in conventional macroscale devices, such as the ability to combine sample collection, analyte extraction, preconcentration, filtration and sample analysis onto a single microfluidic chip [10]. Microfluidics-based lab-on-a-chip devices have received considerable attention in recent years, and are expected to revolutionise clinical diagnostics, DNA and protein analysis and many other laboratory procedures involving molecular biology [11–13]. Currently, most lab-on-a-chip devices employ continuous fluid flow through closed microchannels. However, an alternative approach, which is rapidly gaining popularity, is to mani- pulate the liquid in the form of discrete, unit-sized microdroplets—an approach which is often referred to as digital microfluidics [14–18]. The use of droplet-based microfluidics offers many benefits over continuous flow systems; one of the main advantages is that the reaction or assay can be performed sequentially in a similar manner to traditional (macroscale) laboratory techniques. Consequently, a wide range of established chemical and biological protocols can be transferred to the micro-scale without the need to establish continuous flow protocols for the same reaction.
    [Show full text]
  • Recent Advances in Droplet-Based Microfluidic Technologies
    micromachines Review Recent Advances in Droplet-based Microfluidic Technologies for Biochemistry and Molecular Biology 1, 1, 2, Joel Sánchez Barea y , Juhwa Lee y and Dong-Ku Kang * 1 Department of Chemistry, Incheon National University, Incheon 22012, Korea; [email protected] (J.S.B.); [email protected] (J.L.) 2 Department of Chemistry, Research Institute of Basic Sciences, Incheon National University, Incheon 22012, Korea * Correspondence: [email protected]; Tel.: +82-32-835-8094 These authors contribute equally to this article. y Received: 2 May 2019; Accepted: 18 June 2019; Published: 20 June 2019 Abstract: Recently, droplet-based microfluidic systems have been widely used in various biochemical and molecular biological assays. Since this platform technique allows manipulation of large amounts of data and also provides absolute accuracy in comparison to conventional bioanalytical approaches, over the last decade a range of basic biochemical and molecular biological operations have been transferred to drop-based microfluidic formats. In this review, we introduce recent advances and examples of droplet-based microfluidic techniques that have been applied in biochemistry and molecular biology research including genomics, proteomics and cellomics. Their advantages and weaknesses in various applications are also comprehensively discussed here. The purpose of this review is to provide a new point of view and current status in droplet-based microfluidics to biochemists and molecular biologists. We hope that this review will accelerate communications between researchers who are working in droplet-based microfluidics, biochemistry and molecular biology. Keywords: droplet-based microfluidic; biochemistry; molecular biology; digital PCR; biochip; biosensor; digital quantification; microfluidic; single cell analysis 1.
    [Show full text]
  • Detection of Dna Hybridization on a Configurable Digital Microfluidic
    DETECTION OF DNA HYBRIDIZATION ON A CONFIGURABLE DIGITAL MICROFLUIDIC BIO- CHIP USING SPR IMAGING Lidija Malic1, Teodor Veres2 and Maryam Tabrizian1 1Biomedical Engineering Department, McGill University, CANADA and 2Industrial Materials Institute, National Research Council, CANADA ABSTRACT This paper presents a configurable digital microfluidic-based surface plasmon resonance (SPR) biochip platform comprising an electrowetting-on-dielectric (EWOD) microfluidic device coupled to SPR imaging (SPRi). We demonstrate its application for dynamic on-chip simultaneous immobilization of different DNA probes in combination with multichannel label-free real-time detection of subse- quent hybridization reactions. The integrated EWOD-SPRi system would enable the development of high-throughput, rapid and ultrasensitive biomolecular detection strategies beyond DNA microarray applications. KEYWORDS: Digital microfluidics, EWOD, SPR imaging, DNA hybridization INTRODUCTION EWOD microfluidics has attracted considerable attention in the past decade and the most recent efforts are directed towards its application in biomedical research [1- 3]. While these studies demonstrate the versatility of EWOD devices, the reported applications involve homogeneous phase reactions [4] and detection methods that require labeled biomolecules [2] or sample extraction from the chip [1]. This in- creases both the time and complexity of the assay. To introduce new applications relying on label-free, real-time surface sensitive detection techniques such as SPRi, it would be advantageous to use droplet-based EWOD actuation for surface specific biomolecule immobilization. However, the need of hydrophobic properties for EWOD actuation renders immobilization of biomolecules such as DNA on the sur- face of the chip impossible [3, 4]. In this paper, we demonstrate for the first time the use of an EWOD microfluidic chip to dynamically immobilize DNA probes in a two-dimensional array, followed by SPRi detection of bioaffinity interactions.
    [Show full text]
  • Demonstration of Automated DNA Assembly on a Digital Microfluidic Device
    Rochester Institute of Technology RIT Scholar Works Theses 5-12-2021 Demonstration of Automated DNA Assembly on a Digital Microfluidic Device Hee Tae An [email protected] Follow this and additional works at: https://scholarworks.rit.edu/theses Recommended Citation An, Hee Tae, "Demonstration of Automated DNA Assembly on a Digital Microfluidic Device" (2021). Thesis. Rochester Institute of Technology. Accessed from This Thesis is brought to you for free and open access by RIT Scholar Works. It has been accepted for inclusion in Theses by an authorized administrator of RIT Scholar Works. For more information, please contact [email protected]. ROCHESTER INSTITUTE OF TECHNOLOGY ROCHESTER, NY Demonstration of Automated DNA Assembly on a Digital Microfluidic Device by Hee Tae An A Thesis Submitted in Partial Fulfillment of the Requirements for the Degree of Master of Science in Microelectronic Engineering Submitted May 12, 2021 DEPARTMENT OF ELECTRICAL AND MICROELECTRONIC ENGINEERING KATE GLEASON COLLEGE OF ENGINEERING Approved By: Dr. Michael Schertzer, Associate Professor Date Thesis Advisor, Department of Mechanical Engineering Dr. Karl Hirschman, Professor Date Committee Member, Department of Electrical and Microelectronic Engineering Dr. Ivan Puchades, Assistant Professor Date Committee Member, Department of Electrical and Microelectronic Engineering Dr. Michael Schrlau, Associate Professor Date Committee Member, Department of Mechanical Engineering Dr. Sean Rommel, Director of Microelectronic Engineering/Professor Date Department Representative, Department of Electrical and Microelectronic Engineering Abstract The rapid manufacturing of highly accurate synthetic DNA is crucial for its use as a molec- ular tool, the understanding and engineering of regulatory elements, protein engineering, genetic refactoring, engineered genetic networks and metabolic pathways, and whole-genome syntheses [1,2].
    [Show full text]
  • Protein Stamping for Maldi Mass Spectrometry Using an Electrowetting-Based Microfluidic Platform
    PROTEIN STAMPING FOR MALDI MASS SPECTROMETRY USING AN ELECTROWETTING-BASED MICROFLUIDIC PLATFORM Vijay Srinivasan1, Vamsee Pamula, Phil Paik, and Richard Fair Department of Electrical and Computer Engineering, Duke University, Durham, NC 27708-0291. ABSTRACT MALDI-MS (matrix-assisted laser desorption/ionization mass spectrometry) is one of the most commonly used tech- niques for protein analysis. In conventional systems sample preparation is typically done in well-plates and transferred onto a MALDI target by robotic systems, which are complex, huge, expensive and slow. In this paper, we present a droplet-based microfluidic interface to transfer protein samples from a well-plate format onto a MALDI target for MS analysis. The droplets are actuated using the electrowetting phenomenon, and are immersed in silicone oil which pre- vents non-specific adsorption and enables the manipulation of high concentrations of proteins. Droplet transport and droplet formation were evaluated as a function of protein concentration using bovine serum albumin (BSA) as a test system. Droplet transport was possible for BSA concentrations up to 10mg/mL which is three orders of magnitude higher than previously reported results on handling proteins by electrowetting. Droplet formation from on-chip reser- voirs, using only electrowetting forces and no external pressure assistance, was possible up to concentrations of 0.01mg/mL. An interface between a well-plate format and the electrowetting chip, and a scheme to passively stamp droplets onto a target substrate was then designed and tested by stamping BSA solutions. In two separate experiments 3.6fmoles and 16fmoles of BSA were stamped onto a glass slide using 0.001mg/mL and 0.01mg/mL samples respec- tively.
    [Show full text]
  • Interfacing Microfluidics to LDI-MS by Automatic Robotic Spotting
    Microfluid Nanofluid (2010) 8:777–787 DOI 10.1007/s10404-009-0510-x RESEARCH PAPER Interfacing microfluidics to LDI-MS by automatic robotic spotting Chia-Wen Tsao • Song Tao • Chien-Fu Chen • Jikun Liu • Don L. DeVoe Received: 30 July 2009 / Accepted: 23 September 2009 / Published online: 13 October 2009 Ó Springer-Verlag 2009 Abstract We developed a method of interfacing micro- Keywords Mass spectrometry interfacing Á fluidics with mass spectrometry (MS) using a robotic Microfluidics Á Robotic spotting Á LDI-MS spotting system to automate the contact spotting process. We demonstrate that direct and automated spotting of analyte from multichannel microfluidic chips to a custom 1 Introduction microstructured MALDI target plate was a simple, robust, and high-throughput method for interfacing parallel The ability to perform high-throughput bioanalysis in a microchannels using matrix-assisted laser desorption/ioni- single integrated platform is a substantial benefit offered by zation mass spectrometry (MALDI-MS). Using thermo- microfluidics technology. Arrays of multiplexed micro- plastic cyclic olefin copolymer (COC) polymer microfluidic channels allowing large numbers of simultaneous on-chip chips containing eight parallel 100 lm 9 46 lm micro- analyses have been demonstrated in numerous applications channels connected to a single input port, spotting volume including chemical reaction screening (Wang et al. 2006a), repeatability and MALDI-MS signal uniformity are evalu- immunoassays (Wang et al. 2006b), enzymatic processing ated for a panel of sample peptides. The COC microfluidic (Su et al. 2005), biomolecular separations performed by chips were fabricated by hot embossing and solvent bond- zone electrophoresis (Chen et al. 2002; Shen et al.
    [Show full text]
  • Digital Microfluidics
    Microfluid Nanofluid DOI 10.1007/s10404-007-0161-8 REVIEW Digital microfluidics: is a true lab-on-a-chip possible? R. B. Fair Received: 10 February 2007 / Accepted: 14 February 2007 Ó Springer-Verlag 2007 Abstract The suitability of electrowetting-on-dielectric elemental operations, such as transport, mixing, or dis- (EWD) microfluidics for true lab-on-a-chip applications is pensing. These elemental operations are performed on an discussed. The wide diversity in biomedical applications elemental set of components, such as electrode arrays, can be parsed into manageable components and assembled separation columns, or reservoirs. Examples of the incor- into architecture that requires the advantages of being poration of these principles in complex biomedical appli- programmable, reconfigurable, and reusable. This capa- cations are described. bility opens the possibility of handling all of the protocols that a given laboratory application or a class of applications Keywords Lab-on-a-chip Á Digital microfluidics Á would require. And, it provides a path toward realizing the Biomedical applications Á Detection Á Analysis Á true lab-on-a-chip. However, this capability can only be Electrowetting realized with a complete set of elemental fluidic compo- nents that support all of the required fluidic operations. Architectural choices are described along with the reali- 1 Introduction zation of various biomedical fluidic functions implemented in on-chip electrowetting operations. The current status of Historically, microfluidic devices have been designed from this EWD toolkit is discussed. However, the question re- the bottom up, whereby various fluidic components are mains: which applications can be performed on a digital combined together to achieve a device that performs a microfluidic platform? And, are there other advantages particular application.
    [Show full text]
  • (EWOD): Current Perspectives and Applications in Ensuring Food Safety
    Volume 28 Issue 4 Article 8 2020 Electrowetting-on-dielectric (EWOD): Current perspectives and applications in ensuring food safety Follow this and additional works at: https://www.jfda-online.com/journal Part of the Food Science Commons, Medicinal Chemistry and Pharmaceutics Commons, Pharmacology Commons, and the Toxicology Commons This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License. Recommended Citation Barman, Snigdha Roy; Khan, Imran; Chatterjee, Subhodeep; Saha, Subhajit; Choi, Dukhyun; Lee, Sangmin; and Lin, Zong-Hong (2020) "Electrowetting-on-dielectric (EWOD): Current perspectives and applications in ensuring food safety," Journal of Food and Drug Analysis: Vol. 28 : Iss. 4 , Article 8. Available at: https://doi.org/10.38212/2224-6614.1239 This Review Article is brought to you for free and open access by Journal of Food and Drug Analysis. It has been accepted for inclusion in Journal of Food and Drug Analysis by an authorized editor of Journal of Food and Drug Analysis. Electrowetting-on-dielectric (EWOD): Current perspectives and applications in ensuring food safety REVIEW ARTICLE Snigdha Roy Barman a, Imran Khan b, Subhodeep Chatterjee c, Subhajit Saha a, Dukhyun Choi d,**, Sangmin Lee e,***, Zong-Hong Lin a,c,d,* a Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu 30013, Taiwan b Institute of NanoEngineering and Microsystems, National Tsing Hua University, Hsinchu 30013, Taiwan c Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu 30013, Taiwan d Department of Mechanical Engineering, Kyung Hee University, Yongin, 17104, South Korea e School of Mechanical Engineering, Chung-Ang University, Seoul 06974, South Korea Abstract Digital microfluidic (DMF) platforms have contributed immensely to the development of multifunctional lab-on-chip systems for performing complete sets of biological and analytical assays.
    [Show full text]
  • Lab on a Chip
    Lab on a Chip View Article Online PAPER View Journal On-chip integration of droplet microfluidics and nanostructure-initiator mass spectrometry for Cite this: DOI: 10.1039/c6lc01182a enzyme screening† Joshua Heinemann,ab Kai Deng,ac Steve C. C. Shih,f Jian Gao,b Paul D. Adams,abe Anup K. Singhac and Trent R. Northen*abd Biological assays often require expensive reagents and tedious manipulations. These shortcomings can be overcome using digitally operated microfluidic devices that require reduced sample volumes to automate assays. One particular challenge is integrating bioassays with mass spectrometry based analysis. Towards this goal we have developed μNIMS, a highly sensitive and high throughput technique that integrates drop- let microfluidics with nanostructure-initiator mass spectrometry (NIMS). Enzyme reactions are carried out in droplets that can be arrayed on discrete NIMS elements at defined time intervals for subsequent mass spectrometry analysis, enabling time resolved enzyme activity assay. We apply the μNIMS platform for ki- Received 20th September 2016, netic characterization of a glycoside hydrolase enzyme (CelE-CMB3A), a chimeric enzyme capable of Accepted 2nd December 2016 deconstructing plant hemicellulose into monosaccharides for subsequent conversion to biofuel. This study reveals NIMS nanostructures can be fabricated into arrays for microfluidic droplet deposition, NIMS is com- DOI: 10.1039/c6lc01182a patible with droplet and digital microfluidics, and can be used on-chip to assay glycoside hydrolase enzyme www.rsc.org/loc
    [Show full text]
  • Automated Injection from EWOD Digital Microfluidic Chip Into HPLC Purification System G.J
    Automated injection from EWOD digital microfluidic chip into HPLC purification system G.J. Shah1,2, J. Lei1, S. Chen1, C.-J. Kim1, P.Y. Keng1, R.M. van Dam1 1University of California, Los Angeles, CA 90095, 2Sofie Biosciences, Culver City, CA 90230, USA ABSTRACT We report an automated “chip-to-world” interface between an electrowetting-on-dielectric (EWOD) digital microfluidic device and high-performance liquid chromatography (HPLC) system, expanding the application of EWOD chemical synthesis devices to syntheses that require HPLC purification. The interface collects the crude product from the chip without the need for chip disassembly or other manual intervention. We report (a) bubble-free filling of the injection loop; (b) quantification of how much crude product from the chip is loaded into the loop; and (c) successful injection and HPLC purification of a crude product. It should be noted that because of the small chip volume, analytical-scale HPLC could be used, typically leading to 10-20x more concentrated purified product than semi-preparative HPLC. KEYWORDS Digital microfluidics, Electrowetting on dielectric (EWOD), world-to-chip interface, High-performance liquid chromatography (HPLC), Purification INTRODUCTION Due to its well-controlled volumes, inert surfaces, all-electronic control and flexibility of fluid movement, there has been growing interest in digital microfluidics using electrowetting-on-dielectric (EWOD) for chemical, biochemical and radiochemical synthesis applications [1,5,6]. Several of these require post-synthesis purification, separation and/or analysis. High-performance liquid chromatography (HPLC) is an important and ubiquitous separation technique for both preparative and analytical systems [7]. However, products are often manually pipetted off the chip and introduced into the HPLC [1].
    [Show full text]
  • Digital Microfluidics: a Versatile Platform for Applications in Chemistry, Biology and Medicine
    Digital Microfluidics: A Versatile Platform For Applications in Chemistry, Biology and Medicine By Mais J. Jebrail A thesis submitted in conformity with the requirements for the degree of Doctor of Philosophy – Interdisciplinary Department of Chemistry University of Toronto © Copyright by Mais J. Jebrail (2011) ii Digital Microfluidics: A Versatile Platform For Applications in Chemistry, Biology and Medicnine Mais J. Jebrail Doctor of Philosophy – Interdisciplinary Department of Chemistry University of Toronto 2011 Abstract Digital microfluidics (DMF) has recently emerged as a popular technology for a wide range of applications. In DMF, nL-mL droplets containing samples and reagents are controlled (i.e., moved, merged, mixed, and dispensed from reservoirs) by applying a series of electrical potentials to an array of electrodes coated with a hydrophobic insulator. DMF is distinct from microchannel-based fluidics as it allows for precise control over multiple reagent phases (liquid and solid) in heterogeneous systems with no need for complex networks of microvalves. In this thesis, digital microfluidics has been applied to address key challenges in the fields of chemistry, biology and medicine. For applications in chemistry, the first two-plate digital microfluidic platform for synchronized chemical synthesis is reported. The new method, which was applied to synthesizing peptide macrocycles, is fast and amenable to automation, and is convenient for parallel scale fluid handling in a straightforward manner. For applications in biology, I present the first DMF-based method for extraction of proteins (via precipitation) in serum and cell lysate. The performance of the new method was comparable to that of conventional techniques, with the advantages of automation and reduced analysis time.
    [Show full text]