Download or read book Development of Multi purpose Microfluidic Platform for Single cell and Pairwise Cell Interaction Assay written by Young-Ji Kim and published by . This book was released on 2009 with total page 95 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Cell Analysis on Microfluidics written by Jin-Ming Lin and published by Springer. This book was released on 2017-10-25 with total page 435 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book presents a detailed overview of the design, formatting, application, and development of microfluidic chips in the context of cell biology research, enumerating each element involved in microfluidics-based cell analysis, discussing its history, status quo, and future prospects, It also offers an extensive review of the research completed in the past decade, including numerous color figures. The individual chapters are based on the respective authors' studies and experiences, providing tips from the frontline to help researchers overcome bottlenecks in their own work. It highlights a number of cutting-edge techniques, such as 3D cell culture, microfluidic droplet technique, and microfluidic chip-mass spectrometry interfaces, offering a first-hand impression of the latest trends in the field and suggesting new research directions. Serving as both an elementary introduction and advanced guidebook, the book interests and inspires scholars and students who are currently studying microfluidics-based cell analysis methods as well as those who wish to do so.

Download or read book Microfluidics for Single Cell Analysis written by Jin-Ming Lin and published by Springer Nature. This book was released on 2019-08-28 with total page 263 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book summarizes the various microfluidic-based approaches for single-cell capture, isolation, manipulation, culture and observation, lysis, and analysis. Single-cell analysis reveals the heterogeneities in morphology, functions, composition, and genetic performance of seemingly identical cells, and advances in single-cell analysis can overcome the difficulties arising due to cell heterogeneity in the diagnostics for a targeted model of disease. This book provides a detailed review of the state-of-the-art techniques presenting the pros and cons of each of these methods. It also offers lessons learned and tips from front-line investigators to help researchers overcome bottlenecks in their own studies. Highlighting a number of techniques, such as microfluidic droplet techniques, combined microfluidics-mass-spectrometry systems, and nanochannel sampling, it describes in detail a new microfluidic chip-based live single-cell extractor (LSCE) developed in the editor’s laboratory, which opens up new avenues to use open microfluidics in single-cell extraction, single-cell mass spectrometric analysis, single-cell adhesion analysis and subcellular operations. Serving as both an elementary introduction and advanced guidebook, this book interests and inspires scholars and students who are currently studying or wish to study microfluidics-based cell analysis methods.

Download or read book Multidisciplinary Microfluidic and Nanofluidic Lab on a Chip written by Xiujun (James) Li and published by Newnes. This book was released on 2021-09-19 with total page 486 pages. Available in PDF, EPUB and Kindle. Book excerpt: Multidisciplinary Microfluidic and Nanofluidic Lab-on-a-Chip: Principles and Applications provides chemists, biophysicists, engineers, life scientists, biotechnologists, and pharmaceutical scientists with the principles behind the design, manufacture, and testing of life sciences microfluidic systems. This book serves as a reference for technologies and applications in multidisciplinary areas, with an emphasis on quickly developing or new emerging areas, including digital microfluidics, nanofluidics, papers-based microfluidics, and cell biology. The book offers practical guidance on how to design, analyze, fabricate, and test microfluidic devices and systems for a wide variety of applications including separations, disease detection, cellular analysis, DNA analysis, proteomics, and drug delivery. Calculations, solved problems, data tables, and design rules are provided to help researchers understand microfluidic basic theory and principles and apply this knowledge to their own unique designs. Recent advances in microfluidics and microsystems for life sciences are impacting chemistry, biophysics, molecular, cell biology, and medicine for applications that include DNA analysis, drug discovery, disease research, and biofluid and environmental monitoring. - Provides calculations, solved problems, data tables and design rules to help understand microfluidic basic theory and principles - Gives an applied understanding of the principles behind the design, manufacture, and testing of microfluidic systems - Emphasizes on quickly developing and emerging areas, including digital microfluidics, nanofluidics, papers-based microfluidics, and cell biology

Download or read book Studying Cell Metabolism and Cell Interactions Using Microfluidic Devices Coupled with Mass Spectrometry written by Huibin Wei and published by Springer Science & Business Media. This book was released on 2012-10-17 with total page 117 pages. Available in PDF, EPUB and Kindle. Book excerpt: This thesis describes a new approach for cell analysis by the rapid developing microfluidic technology. The nominee has made great contributions to develop a new analysis platform which combined microfluidic devices with mass spectrometry to determine the trace compounds secreted by cells. Based on this analysis platform, she studied the specific cell secreting behaviors under controlled microenvironment, of which the secretion compounds were qualified and semi-quantified by mass spectrometry. A novel cell sorting device integrated homogenous porous PDMS membrane was invented to classify cells from real samples based on the size difference. The nominee further studied the signal transmission between different cells, and the signal chemicals were qualitative and quantitative monitored by the analysis platform. This indicates the potential significant application of the new cell analysis platform in medicine screening and early diagnosis.

Download or read book Single Cell Assays written by Paul C.H. Li and published by Springer Nature. This book was released on 2023-07-10 with total page 227 pages. Available in PDF, EPUB and Kindle. Book excerpt: This detailed volume explores the use of single-cell assays in research for drug discovery, microfluidics, and more. The book delves into methodologies involving a variety of cell types and diseases, small molecules and biologics, as well as studies of the genome and transciptome. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Single-Cell Assays: Microfluidics, Genomics, and Drug Discovery serves to enable researchers to obtain a rapid overview in state-of-the-art microfluidic single-cell assays and an impression of what possibilities these assays offer to drug discovery.

Download or read book Microfluidic Platform for Multi parameter Analysis of Live Single Cells written by Meryl Rodrigues and published by . This book was released on 2014 with total page 54 pages. Available in PDF, EPUB and Kindle. Book excerpt: Cellular heterogeneity is a key factor in various cellular processes as well as in disease development, especially associated with immune response and cancer progression. Cell-to-cell variability is considered to be one of the major obstacles in early detection and successful treatment of cancer. Most present technologies are based on bulk cell analysis, which results in averaging out the results acquired from a group of cells and hence missing important information about individual cells and their behavior. Understanding the cellular behavior at the single-cell level can help in obtaining a complete profile of the cell and to get a more in-depth knowledge of cellular processes. For example, measuring transmembrane fluxes oxygen can provide a direct readout of the cell metabolism. The goal of this thesis is to design, optimize and implement a device that can measure the oxygen consumption rate (OCR) of live single cells. A microfluidic device has been designed with the ability to rapidly seal and unseal microchambers containing individual cells and an extracellular optical oxygen sensor for measuring the OCR of live single cells. The device consists of two parts, one with the sensor in microwells (top half) and the other with channels and cells trapped in Pachinko-type micro-traps (bottom half). When the two parts of the device are placed together the wells enclose each cell. Oil is flown in through the channels of the device to produce isolated and sealed microchamber around each cell. Different fluids can be flowed in and out of the device, alternating with oil, to rapidly switch between sealed and unsealed microenvironment around each cell. A fluorescent ratiometric dual pH and oxygen sensor is placed in each well. The thesis focuses on measuring changes in the oxygen consumption rate of each cell within a well. Live and dead cells are identified using a fluorescent live/dead cell assay. Finally, the technology is designed to be scalable for high-throughput applications by controlling the flow rate of the system and increasing the cell array density.

Download or read book Development of a Microfluidic Platform for Multicellular Tumour Spheroid Assays written by Kay Seonaid McMillan and published by . This book was released on 2016 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Microfluidics is a valuable technology for a variety of different biomedical applications. In particular, within cancer research, it can be used to improve upon currently used in vitro screening assays by facilitating the use of 3D cell culture models. One of these models is the multicellular tumour spheroid (MCTS), which provides a more accurate reflection of the tumour microenvironment in vivo by reproducing the cell to cell contact, the development of a nutritional gradient and the formation of a heterogeneous population of cells. Therefore, the MCTS provides a more physiologically relevant in vitro model for testing the efficacy of treatments at the preclinical level. Currently, methods for the formation and culture of spheroids have several limitations, including being labour intensive, being low throughput, producing shear stress towards cells and the hanging drop system being unstable to physical shocks. Recently, microfluidics (especially droplet microfluidics) has been employed for the culture and screening of spheroids, providing a high-throughput methodology which only requires small volumes of fluids and small numbers of cells. However, current issues with droplet microfluidics include complicated droplet gelation procedures and short cell culture times.In this thesis, the use of microfluidic technologies as an approach for spheroid formation and culture are investigated with the aim to create a platform for radiotherapeutic and chemotherapeutic treatment of spheroids using cell lines. Initially, the use of emulsion technology at the macro scale was evaluated to determine the best conditions for spheroid culture. Once this was achieved the spheroids were compared to spheroids using a traditional method and radiotherapeutic treatment was conducted. Subsequently, avenues for miniaturising the developed emulsion-based methods were studied to provide a microfluidic technology. Finally, along with identifying the optimal culture conditions using hydrogels, a microfluidic system that integrated both droplet and single phase microfluidics features was developed for the formation and culture of spheroids. Using the latter, proof of principle experiments were conducted to demonstrate the suitability of the platform for both chemotherapeutic and radiotherapeutic assays within the same device.

Download or read book High Throughput Microfluidic Technologies for Cell Separation and Single cell Analysis written by Lidan Wu (Ph. D.) and published by . This book was released on 2016 with total page 102 pages. Available in PDF, EPUB and Kindle. Book excerpt: The heterogeneity of individual cellular behavior in response to physical and chemical stimuli has raised increasing attention in many biological processes. There is great incentive in developing techniques for high throughput single-cell measurements and manipulations. Particularly, cell size has been recognized as an important parameter in single cell study and pericellular protease activity plays a key role in regulating the microenvironment of individual cells. Therefore, this thesis focuses on establishing new methods to address the issues of cell size and single cell protease measurement. We first develop a size-based cell separation technique using Dean-coupled inertial microfluidic sorter. Separation of cells by size before downstream assays might be beneficial in simplifying the system and facilitating the discovery of rare subpopulations through enrichment of cells with certain size range or cell cycle phase. By investigating the particle focusing and separation mechanisms in curved microfluidic channel, we develop a novel design of inertial microfluidic sorter with higher separation resolution and then demonstrate its capacity in leukocyte isolation from blood. This novel cell sorter would be a promising alternative to many other cell separation problems. We then establish a microfluidic platform for functional measurement of single cell pericellular proteases, including both those secreted and expressed on cell surface. We apply the platform to studying the PMA-mediated protease response of HepG2 cells at single-cell level and reveal the diversity in the dynamic patterns of single-cell protease activity profile upon drug stimulation. We also present the preliminary exploration of single-cell protease activity behavior in anticancer drug resistance development. Lastly, we explore the applicability of our platform for single-cell shedding measurement. Protease-mediated molecular shedding is one of the key mechanisms through which individual cells actively regulate their own microenvironment. However, the amount of molecules being shed for individual cells is extremely low, posing significant challenges in detecting shedding quantitatively. By means of analytical analysis and numerical simulations, we investigate the intrinsic noise of low-abundance molecule detection. Experimental characterizations have also been performed to evaluate the impact of practical factors on actual readout variation.

Download or read book Biomimetic Microfluidic Platforms for Monitoring Cellular Interactions in Microscale Flow written by Erdem Kucukal and published by . This book was released on 2020 with total page 175 pages. Available in PDF, EPUB and Kindle. Book excerpt: Cell-cell and cell-protein interactions strongly regulate critical cellular processes such as cell differentiation, cell proliferation, and cell division. To have a better understanding of these interactions, micro-engineered biomimetic platforms providing physiologically relevant environments are needed. Last decades have witnessed remarkable advances in micro/nanotechnologies, through which novel micro-platforms have been developed to mimic physiological microenvironments, providing a means to better understand cellular biomechanics, such as adhesive interaction between different cell types, in vitro. These assays have remarkably enhanced our understanding of many different pathophysiologies that stem from altered cellular mechanical properties as in cancer and sickle cell disease (SCD). The adhesive and deforming characteristics of red blood cells (RBCs) play a critical role in vascular occlusions that lead to life-threatening crises in SCD. The altered properties of sickle hemoglobin (HbS) containing RBCs reduce cellular deformability and increase cellular adhesion onto extracellular matrix (ECM) proteins as well as endothelial cells. This sophisticated dynamic process takes place within a wide range of shear rates indigenous to different types of microvasculature. However, most microfluidic in vitro adhesion assays either do not mimic the shear rate transitioning or they do so through discreet shear rate alterations rather than a continuous transition, which fails to fully recapitulate the flow dynamics in the microvasculature. In Chapter 2, a shear-gradient microfluidic device that creates a variable shear gradient along the flow direction to investigate the adhesion of RBCs under continuously transitioning shear rates is developed and functionalized with endothelium associated biomolecules. With this method, shear gradient dependent adhesion of RBCs from healthy and SCD subjects are quantified, and level of this adhesion is categorized by new parameters such as Shear Gradient Microfluidic Adhesion (SiGMA) Index and Adhesion Threshold (AT). Heterogeneity in shear gradient dependent adhesion levels are reported and correlated with subject clinical phenotype such as increased inflammation and iron overload. Cell-ECM protein interactions represent the environment in which endothelial cell damage leads to the exposure of sub-endothelium with which circulation blood cells can interact. This sort of damage may be triggered in certain pathological scenarios such as SCD. Nevertheless, a more complete picture should also include endothelial cells to better mimic the intravascular microenvironment in studying cellular interactions. In Chapter 3, a microfluidic system integrated with cultured human umbilical vein endothelial cells (HUVECs) is developed. Patients with SCD usually has ongoing in vivo hemolysis, and as a result, the free heme levels in their plasma significantly increase because of accelerated RBC lysis. To simulate this favored hemolytic microvascular conditions observed in SCD, the endothelial monolayer is treated with heme and its effect on RBC adhesion is assessed. Furthermore, the associations between heme-mediated RBC adhesion and subject clinical variables (lactate dehydrogenase (LDH), reticulocyte counts, age, etc.) are unraveled. Cellular adhesive interactions between blood cells, combined with altered mechanical properties, can manifest in altered biophysical characteristics of whole blood as a tissue. One of the implications is abnormal blood viscosity that can lead to lethal cardiovascular complications. In Chapter 4, a microfluidic platform integrated with the micro particle image velocimetry is developed to probe whole blood viscosity (WBV) in subjects with SCD. The associations between WBV and RBC adhesion to an endothelium associated biomolecule as well as between WBV and subject clinical variables, such as hematocrit and RBC count, are reported. In Chapter 5, a tube-in-tube flow-loop platform, similar to a system commonly utilized in industrial applications, is first developed to analyze the hemocompatibility of four different biomaterials used in vascular catheter manufacture. Then a novel microfluidic flow-loop assay is designed and fabricated for the same purpose. Non-fouling characteristics of these materials are assessed via a protein binding assay, in which fluorescently labeled human fibrinogen and bovine serum albumin was flowed over the biomaterial surface under physiologic flow conditions. Non-thrombogenicity is quantified by flowing whole bovine blood over the biomaterials followed by an acid phosphatase colorimetric protocol to infer platelet adhesion / thrombus formation at the cell-biomaterial interface. Results from the microfluidic device are compared against the tube-in-tube platform, and the hemocompatibility of these four biomaterials are discerned.

Download or read book Developing Droplet Microfluidic Technologies for Single cell and Single molecule Assays written by Soroush Kahkeshani and published by . This book was released on 2017 with total page 115 pages. Available in PDF, EPUB and Kindle. Book excerpt: In the past decade, growing number of commercialized "lab on a chip" systems have been addressing needs in biology for patients and researchers. The ability to miniaturize conventional systems used in lab while increasing sensitivity in addition to newly developed platforms, have enabled new applications such as point-of-care diagnostics and single-cell sequencing. This dissertation focuses on new droplet microfluidic platforms developed for addressing the current challenges (controlling encapsulation beyond Poisson distribution, removing bulky and expensive pumps,...) in commercialization of single-cell and single-molecule assays. In chapter 1, current state of the art, recent commercialized products and their limitations are discussed. In chapter 2, new findings about particle interactions in single-phase microfluidics leading to "preferred interparticle spacings" and trajectory attractors in inertial microflow regimes are presented. Furthermore, these findings have been combined with new modifications made to conventional flow focusing droplet generators in order to avoid jetting regime in inertial regimes, thus allowing making use of preferred spacing between cells/particles in inertial flows for controlling number of cells/particles encapsulated in droplets (one cell/particle per drop). Our findings could be used for many applications such as drug discovery. The other application could be for droplet sequencing (drop-seq) where for example RNA of individual cells are sequenced and it is important that only one cell is present in each droplet to generate unbiased data. In chapter 3 and 4 of the dissertation, new patented technology for generation of droplets, replacing bulky and expensive pumps, with body force exerted on the ferrofluid by magnetic field acting on magnetic nanoparticles mixed with the fluid. Thus the cost and footprint of the device is reduced significantly, allowing for use in limited-resource settings (global health) for quantification of biomolecules by controlled magnetic dosing of nanoliter droplets.

Download or read book Integrated Microfluidic Device for Single cell High Throughput Screening in Dynamic Gene Expression Analysis written by Lawrence Kwan Yeung Hui and published by . This book was released on 2008 with total page 60 pages. Available in PDF, EPUB and Kindle. Book excerpt: Over the past decade, interest in microfluidics has surged as applications have trended towards novel biological assays. Specifically, the ability of microfluidics to parallelize cellular studies through array-based chip designs has attracted researchers interested in investigating cellular function under a wide variety of environmental conditions. The capability of microfluidic devices to control microenvironment conditions and induce dynamic perturbation to cellular systems makes microfluidics (or "lab-on-a-chip") an attractive platform to study gene expression dynamics. In this project, the functionality of microfluidic technology is exploited to design and construct a device for isolation and observation of cells in high throughput. The integration of a concentration gradient with homogenous medium within each chamber was designed specifically to investigate gene regulation in Saccharomyces cerevisiae under various concentrations of chemical inducers. These devices were designed to sustain cells for extended periods of time with high temporal resolution to study dynamic gene expression in single cells. The device builds on previous studies by probing up to eight distinct cell cultures in parallel. The microfluidic platform was then used to study yeast cells at various levels of inducer perturbations. Further experimentation revealed the utility of a parallel gradient by producing an induction curve of the yeast response. Such high-throughput designs will prove essential to yeast systems biology research as it strives to understand the complex regulatory interactions that dictate cell function by probing vast regions of parameter space.

Download or read book New Microfluidic Platforms for Cell Studies written by Irena Barbulovic-Nad and published by . This book was released on 2009 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book A Microfluidic Platform for the Genome wide Analysis of Electrical Phenotype written by Michael David Vahey and published by . This book was released on 2010 with total page 143 pages. Available in PDF, EPUB and Kindle. Book excerpt: This thesis presents the development of a new microfluidic method for separating and characterizing cells based upon differences in their electrical properties. The objective of this work is to obtain a genome-wide mapping of genotype to electrical phenotype in the budding yeast Saccharomyces cerevisiae. Towards this end, we present: (1) the development of a novel equilibrium gradient separation method, called isodielectric separation (IDS); (2) the development of physical theories describing how interactions between particles effect microscale separations; and (3) the application of IDS to a screen for electrical phenotypes in the yeast deletion library. Despite the variety of technologies available for cell sorting, a relative lack of intrinsic separation methods - those which separate cells according to their natural, unmodified characteristics - persists. To address this need, we have developed isodielectric separation. IDS separates cells according to differences in their intrinsic electrical properties. Using dielectrophoresis in a medium with spatially varying electrical conductivity, IDS drives cells to the locations where their polarization charges vanish, spatially resolving cells with different electrical properties. Our implementation of IDS offers label-free, continuous-flow separation, and is capable of resolving graded differences in electrical properties. Additionally, we demonstrate the ability to extract quantitative information from samples during separation, establishing IDS as an analytical technique as well as a preparative one. Any platform for performing genetic screens must have high throughput. Although satisfying this requirement would be greatly facilitated by using high cell densities, physical interactions between cells under these conditions can affect the performance of devices used for screening. Although pervasive, interactions between cells or particles are challenging to describe quantitatively, especially in the confined environments typical of microfluidic devices. By studying the effects of electrostatic and hydrodynamic interactions between particles in a microfluidic device, we have found that ensembles of interacting particles exhibit emergent behaviors that we are able to predict through numerical simulations and a simple analytic model based on hydrodynamic coupling. Applying these theoretical tools to IDS and other microfluidic separation methods has provided insight into how particle interactions can profoundly influence separation performance in counterintuitive ways. Having demonstrated the performance metrics necessary for a genetic screen, we apply IDS to the genome-wide analysis of electrical properties in the budding yeast S. cerevisiae. Although others have studied changes in electrical properties induced by drastic changes in gene expression (e.g. in differentiation) or by specific mutations in a small number of genes, a systematic and comprehensive analysis of the relationship between genotype and electrical phenotype has yet to be performed. Using IDS, we have screened the -5000 strains in the yeast deletion collection for altered electrical phenotype. This work has identified a number of genes associated with distinct electrical properties, and, by analyzing known interactions and correlations between these genes, we have identified pathways and morphologies that appear to be primary determinants of electrical phenotype.

Download or read book Development of a Novel Microfluidic Platform to Study T Cell Signaling written by Shannon L. Faley and published by . This book was released on 2007 with total page 109 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Development of an Automated 3D Flow focusing Microfluidic Platform for Single Cell Sorting written by Yingkai Lyu and published by . This book was released on 2020 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Development and Application of a Microfluidic Platform for Single cell Cultivation of CHO Suspension Cell Lines written by Julian Schmitz and published by . This book was released on 2022 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: