SELECTBIO

Lab on a Chip World Congress 2013

There is a strong drive within the forensic science community to improve the speed, cost, portability, and reliability of forensic DNA evidence processing.  This path toward integration will allow for rapid suspect identification and detention at the point of contact.  The authors present an integrated DNA analysis platform for fully automated “swab-in profile-out” evidence processing.  This self contained device eliminates the need for highly trained forensic personnel and reduces the possibility of run-to-run contamination.  The authors demonstrate automated workflow processing of solid-phase biological samples on a real world size scale; quickly extracting genetic biomaterials, performing 17-plex STR-PCR amplification, and delivering the amplicons to downstream multiplexed CE detection.  The unique hurdles associated with obtaining rapid high quality genetic analysis across multiple loci and sample types are discussed and recent developments are presented.


Advancements in Automated Sample Preparation, STR-PCR, and CE Detection for Rapid Nucleic Acid Analysis on an Integrated Microfluidic Platform

The greatest challenge in implementing complex bioanalytical assays on a chip-sized platform is to develop a fluid control system that is simple yet fully functional. Manipulation of droplets on an open surface promises easier, more flexible, and more functionally integrated liquid control, than does continuous flow microfluidics. The talk focuses on the development of a droplet microfluidic platform for the detection of disease biomarkers and infectious pathogens using crude biosamples. The platform exploited the dual functionality of silica superparamagnetic particles (SSP) for solid phase extraction of DNA and magnetic actuation. This enabled the integration of sample preparation and genetic analysis within discrete droplets, including the steps of cell lysis, DNA binding, washing, elution, amplification and detection. The microfluidic device was self contained, with all reagents stored in droplets, thereby eliminating the need for fluidic coupling to external reagent reservoirs. In addition, a compact sample handling stage, which integrated the magnet manipulator, the droplet microfluidic device and a Peltier thermal cycler, was built for convenient droplet manipulation and real-time detection. Feasibility of the platform was demonstrated by analysing ovarian cancer biomarker Rsf-1 and detecting infectious pathogen with the real time polymerase chain reaction.


All-in-one Droplet Microfluidic Chips for Biomolecular Assays

Commercialization of Microfluidic Devices for Point-of-Care Diagnostics

The use of continuous flow reactors for gas/liquid chemical reactions remains relatively undeveloped even though application of such devices for reactions involving highly reactive gases may, potentially, be highly beneficial. Our long term interest in the synthesis of molecules bearing fluorine atom substituents by selective transformation of carbon-hydrogen bonds to carbon-fluorine bonds using highly reactive elemental fluorine, prompted us to develop flow reactor systems for the purpose of increasing selectivity, controllability and safety of direct fluorination processes.   The development and use of practical, easily maintained, economic and simple multi-channel thin film gas/liquid flow reactors that may be fabricated using standard workshop techniques will be described. Direct fluorination of 1,3-dicarbonyl systems on a practical scale (e.g. 100 g per 24 hour period in a 9-channel device) demonstrates the applicability of flow devices for industry.   Recent use of the HOF.MeCN complex, formed by reaction of fluorine with water in acetonitrile, for various oxidation processes will also be presented.

Continuous Flow Reactors for Gas/Liquid Chemical Processes

Microfluidic systems have gained popularity as key experimental tool in the chemical and biological sciences due to a range of features. These include the ability process small reagent/sample volumes, enhanced analytical performance, reduced instrumental footprint, low unit cost, facile integration of functional components within monolithic substrates and the capacity to exploit atypical fluid behaviour to control chemical and biological operations in both time and space. Droplet-based or segmented-flow microfluidics exploits the spontaneous formation of droplets within microchannel environments to perform a variety of analytical processes. These droplets have sub-nanoliter volumes and act as an individual, isolated reaction vessels. Variation of the cross-sectional dimensions of microchannels can be used to regulate droplet volumes, and flow rate variation allows control of reagent concentrations. Importantly, droplets can be generated at kHz frequencies, meaning that millions of individual reactions can be processed in very short times. In my talk I will report recent advances in this area with particular reference to high throughput reaction screening and cell-based assays.



Droplet-Based Microfluidics for High-Throughput Biological Screening

Intercellular communication is strongly dependent on the spatial organization of tissues at the micrometer scale. We present microfabricated cell culture substrates that enable fine-grained control of cell positioning, allowing the investigation of contact-dependent signaling dynamics, short-range paracrine effects, and feedback signaling. Reconfigurable comb substrates allow two cell populations to be co-cultured either in direct contact or close proximity without contact; pure populations can also be quickly recovered from the co-culture. The system was originally developed for the study of cell-cell interactions in the liver, where we discovered the importance of direct contact as well as the temporal length of contact. More recently, we have discovered short-range paracrine effects in tumor-stromal culture and ES-feeder culture. In the cancer system, this provides a platform for high-throughput profiling of effects from tumor-stromal crosstalk. In the stem cell system, this provides an unmixed feeder-layer system for maintaining pluripotency without contamination of the ES cells by the feeders. 


Investigation of Cell-cell Interactions via Compartmentalized Co-culture Platforms

Chemical gradients drive many processes in biology, ranging from nerve signal transduction to ovulation. At present, microscopy is the primary tool used to understand these gradients. Microscopy has provided many important breakthroughs in our understanding of the fundamental biology, but is limited due to the need to incorporate fluorescent molecules into biological systems. As a result, there is a need to develop tools that can measure chemical gradient formation in biological systems that do not require fluorescent modification of the targets in question, can be multiplexed to measure more than one molecule and is compatible with a variety of biological sample types, including in vitro cell cultures and ex vivo tissue slices. Work from our group on the development of microfluidic tools to measure chemical gradients in living tissue will be presented. Two separate systems are under development. The first is a microfluidic system designed to analyze metabolite and protein expression from tissue. The sampling system can resolve up to 19 different ports and can be interface with either electrochemical or fluorescence-based detection methods. Using these two detection methods, we are capable of analyzing the release of either small molecule metabolites or proteins and peptides using immunoassays. The second system uses a high-density electrode array to image release of electrochemically active metabolites like nitric oxide from live tissue slices. Electrochemical characterization of this system combined with a microfluidic system for gradient generation will be shown.


Mapping Chemical Gradients in Living Tissue in Space and Time Using Microfluidics

In this work, we report on an in vitro systematic fluid dynamic investigation of RBC suspensions flowing in a microcirculation-mimicking silicon device containing a network of microchannels of diameter comparable to cell size. The microfluidics device is made of PDMS and is fabricated by using soft-lithography techniques. RBC membrane rheological behavior is investigated by analyzing the transient behavior of RBC shape in confined flow and by measuring the membrane viscoelastic properties in converging/diverging microchannels. The time scale of the transient process is independent on the applied pressure drop and the measured value for healthy cells is in agreement with previous micropipette data from the literature. In order to study the effect of reduced deformability of the RBC membrane, the flow behavior of glutaraldehyde (GA)-hardened RBCs has been also analyzed. The comprehension of the single cell behavior leaded to the analysis of the RBC flow-induced clustering. Cluster size and velocity is investigated as a function of the applied pressure drop and of the microcapillary residence time, including the effect of polydispersity. The main results of this work is the development of a novel methodology to estimate cell membrane viscoelastic properties. Possible applications include the analysis of RBC deformability in pathological situations, for which reliable quantitative methods are still lacking.   


Microconfined Flow-based Imaging Methods to Study Red Blood Cell Deformability and Clustering in Vitro

"Z-microsystems is a 60 years tooling and injection molding company. Since 13 years we are a reliable partner for our customers in the medical market. In this technology spotlight we give you insights into thin channels, chambers and variation of different thicknesses are possible to manufacture in several forms of e.g. chips, cartridges, micro arrays with optical combination and so on. The one of the main Advantages is the roughness smooth and precise Nano- or Microstructures with small tolerances to build up in the insert and in the injection molded parts too. We are ISO 9001:2008 &amp; ISO 13485:2003 + AC:2007 certified.
Processes: Fluidic development &gt; Prototyping &gt; Pre Series &gt; Series &gt; Bonding &gt; Coating
"


More Accuracy & Precision, Through Tighter Tolerances & Smoother Microfluidic Channels - A Case Study

Smart Consumables with microscale or supreme optical features are prerequisites for emerging applications in the biomedical markets. The increasing complexity of such new products requires new manufacturing technologies. Sony DADC BioSciences offers development, manufacture and supply of polymer-based smart consumables to OEM partners. Specializing in customized mass manufacturing of highly sophisticated consumables, Sony DADC actively applies expertise in innovation to offer state-of-the-art solutions to the biomedical market. Smart polymer-based consumables have been created to solve a variety of experimental assay problems. Applications in high-throughput screening in genomics, in-vitro diagnostics, and single cell assays will be presented.


New Generation Polymeric Materials Leveraging Nanostructuring and Coating Technologies

In April 2003, the Human Genome Project was declared complete. This first human genome sequencing took 13 years, mobilized 20000 researchers and cost $3B! Since the early 2000s, sequencing technologies have come a long way from Sanger sequencing (also called 1st generation sequencing) used for the Human Genome Project to the Next Generation Sequencing technologies available today.  Next Generation Sequencing (NGS) technologies have demonstrated the capacity to sequence DNA at unprecedented speed, enabling previously unimaginable novel healthcare applications. Indeed, by sequencing a large number of DNA molecules in parallel after PCR amplification, technology developers have been able to sharply increase machine throughput by a factor of 10. The launch of a new detection method based on electrochemical detections was also a huge step forward for the sequencing world. At the same time, price has taken a steep decline.  Starting its transition from research to clinical market, the number of NGS Platforms used worldwide will surge in the next several years and the run time will continue to decrease. This will make the NGS Consumables market explode. Yole Développement foresees the NGS Consumables market growing to $3.1B in 2018, the result of several millions of cartridges being sold each year.  This presentation will describe both market and technology trends of the NGS market.


Next Generation Sequencing Chips: Market & Technology Trends

Leveraging the inherent heat-induced relaxation of pre-stressed thermoplastic sheets - commodity shrink-wrap film - we pattern in a variety of ways at the large scale and achieve our desired structures by controlled shrinking down to 5% of the original, patterned sizes. This enables us to 'beat' the limit of resolution inherent to traditional 'top-down' manufacturing approaches. With these tunable shape memory polymers, we can robustly integrate extremely high surface area and high aspect ratio nanostructures directly into our microsystems. 


Shrink Manufacturing Advanced Research Tools (SMART) 

We are investigating tunable wetting of a smart polymer - dodecylbenzenesulfonate-doped polypyrrole (PPy(DBS)). Here, we are probing fundamental mechanisms related to the electrochemical transport and reversible wetting states of PPy(DBS) upon continuous reduction and oxidation reactions. Successful research on this tunable wetting concept will impact a wide range of microfluidics applications, attributed to the low-voltage operation. 


Smart Polymer-based Microfluidics

Microfluidics has in recent years made the critical transition from being a technology looking for a problem to a real problem-solving tool. However a comparatively high entry barrier still exists for the utilization of this technology due to its multidisciplinary nature and apparent complexity. Especially in the development of highly integrated sample-in-result-out devices e.g. for molecular diagnostics or biothreat analysis, the generation of a successful microfluidic design de-novo often proves to be a significant challenge. In order to solve this problem, we have developed a modular toolbox approach. In this strategy, functional elements are defined, realized and validated and combined in the second stage to an integrated device. We will present examples of such developments for applications in molecular diagnostics, food and pathogen analysis, and point out critical stages of such developments as well as insights in overcoming these hurdles. By modularizing developments, especially the early proof-of-concept phases can be realized faster and at lower cost compared to conventional approaches. 


Advancements in Automated Sample Preparation, STR-PCR, and CE Detection for Rapid Nucleic Acid Analysis on an Integrated Microfluidic Platform

Posted by

SELECTBIO

4,690 views

|

October 16, 2013

Lab on a Chip World Congress 2013

Keep up to date with all your favourite videos and channels.

Get personalised notifications on new releases and channel content by subscribing to the LabTube eNewsletter.