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Full-Text Articles in Physical Sciences and Mathematics
Synthesis Of Thiol-Acrylate Hydrogels For 3d Cell Culture And Microfluidic Applications, Anowar Hossain Khan
Synthesis Of Thiol-Acrylate Hydrogels For 3d Cell Culture And Microfluidic Applications, Anowar Hossain Khan
LSU Doctoral Dissertations
Globally cell culture is an $18.98 billion industry as of 2020, with an 11.6 percent annual growth rate. Drug discovery has an estimated worth of $69.8 billion in 2020 and is predicted to grow to $110.4 billion by 2025. Three-dimensional (3D) cell culture of cancer cells is one of the rapidly growing felids since it better recapitulates in vivo conditions compared to two-dimensional (2D) models. However, it is challenging to grow 3D tumor spheroids outside the body, and some of the existing technology can generate these spheroids outside the human body but poorly mimic in vivo tumor models. Therefore, there …
Thiol-Acrylate Polymerization Kinetics And Applications In Microfluidics, Michael Perrin Tullier
Thiol-Acrylate Polymerization Kinetics And Applications In Microfluidics, Michael Perrin Tullier
LSU Doctoral Dissertations
The reaction kinetics and some applications in the field of microfluidics for thiol-acrylate Michael addition polymerizations using multifunctional monomers have been researched and are presented here. The polymerization rate constants for base-catalyzed systems were found to increase with increasing thiol and acrylate functionality, which was attributed to the intramolecular interactions between functional groups. The nucleophile-initiated thiol-acrylate Michael addition polymerization kinetics were monitored via FTIR, and it was determined that the increase in the rate of reaction in these multifunctional systems was significantly less dramatic than the increase observed in monofunctional systems. While no radical polymerization was observed during most typical …
Microfluidic And Nanofluidic Devices - Mixed Scale Systems For Bioanalytical Applications, Michael David Vincent
Microfluidic And Nanofluidic Devices - Mixed Scale Systems For Bioanalytical Applications, Michael David Vincent
LSU Doctoral Dissertations
Developments in the field of microfluidics and nanofluidics allow an experimenter to employ new analytical techniques. One direction in which the field is moving towards is developing new devices and or instruments on the smallest scale possible. One way in which this can be achieved is by using a combination of top-down and bottom-up technologies. Some of these top-down techniques have been used in other fields such as the use of photolithography to fabricate semiconductors. Photolithography is a top-down method used to create features normally on the micrometer scale, but in order to create nanometer size features another top-down method …
Polymer Micro- And Nanofluidic Systems For In Vitro Diagnostics: Analyzing Single Cells And Molecules, Swathi Reddy Pullagurla
Polymer Micro- And Nanofluidic Systems For In Vitro Diagnostics: Analyzing Single Cells And Molecules, Swathi Reddy Pullagurla
LSU Doctoral Dissertations
Polymer micro- and nanofluidic systems, with their critical dimensions, offer a potential to outperform conventional analysis techniques and diagnostic methods by enhancing speed, accuracy, sensitivity and specificity. In this work, applications of microfluidics have been demonstrated to address the existing challenges in stroke diagnosis, by mRNA expression profiling from whole blood within <20 min. A brief overview of various biomarkers for stroke diagnosis is given in chapter 1 followed by design and testing of individual microfluidic modules (chapter 2 and 3) required for the development of POC diagnostic strategy for stroke. We have designed and evaluated the performance of polymer microfluidic devices for the isolation of leukocyte subsets, known for their differential gene expression in the event of stroke. Target cells (T-cells and neutrophils) were selected from with greater purities, from 50 µl whole human blood by using affinity based capture in COC devices within a 6.6 min processing time. In addition, we have also demonstrated the ability to isolate and purify total RNA by using UV activated polycarbonate solid phase extraction platform. Polymer-based nanofluidic devices were used to study the effects of surface charge on the electrodynamic transport dynamics of target molecules. In this work, we report the fabrication of mixed-scale micro- and nanofluidic networks in poly(methylmethacrylate), PMMA, using thermal nanoimprint lithography using a resin stamp and surface modification of polymer nanoslits and nanochannels for the assessment of the associated electrokinetic parameters – surface charge density, zeta potential and electroosmotic flow. This study provided information on possible routes that can be adopted to engineer proper wall chemistry of polymer nanochannels for the enhancement or reduction of solute/wall interactions in a variety of relevant single-molecule studies.
A Microfluidic Bacteria Culturing Device With Maldi Mass Spectrometry Detection, Pratap Reddy Machavaram Siva
A Microfluidic Bacteria Culturing Device With Maldi Mass Spectrometry Detection, Pratap Reddy Machavaram Siva
LSU Master's Theses
A novel microfluidic device was developed for bacterial cell culturing using mass spectrometry as the detector. One of the challenges in proteomics is to achieve high sensitivity in the identification of proteins in complex samples with widely varying concentrations. The main limitations for proteomic studies are relatively slow and labor-intensive steps such as cell culturing and protein digestion of small sample quantities. Microfluidics is a promising approach to increase throughput and to reduce the time-consuming steps that are necessary for proteomics. When an analytical detection method is combined with microfluidics it can overcome limitations that are important in the analysis …
Microfluidics For The Analysis Of Integral Membrane Proteins: A Top-Down Approach, Katrina Nychole Battle
Microfluidics For The Analysis Of Integral Membrane Proteins: A Top-Down Approach, Katrina Nychole Battle
LSU Doctoral Dissertations
The development of fully automated and high-throughput systems for proteomics is now in demand because of the need to generate new protein-based disease biomarkers. Unfortunately, it is difficult to identify protein biomarkers that are low abundant when in the presence of highly abundant proteins, especially in complex biological samples like serum, cell lysates, and other biological fluids. Membrane proteins, which are in many cases of low abundance compared to cytosolic proteins, have various functions and can provide insight into the state of disease and serve as targets for new drugs making them attractive biomarker candidates. Traditionally, proteins are identified through …
Fully Integrated Microsystem For Bacterial Genotyping, Hui-Wen Chen
Fully Integrated Microsystem For Bacterial Genotyping, Hui-Wen Chen
LSU Doctoral Dissertations
Methods for bacterial detection and identification has garnered renewed interest in recent years due to the infections they may cause and the antimicrobial resistances they can develop, the potential for bioterrorism threats and possible contamination of food/water supplies. Therefore, the rapid, specific and accurate detection of pathogens is crucial for the prevention of pathogen-related disease outbreaks and facilitating disease management as well as the containment of suspected contaminated food and/or water supplies. In this dissertation an integrated modular-based microfluidic system composed of a fluidic cartridge and a control instrument has been developed for bacterial pathogen detection. The integrated system can …
Selection And Enumeration Of Low Epcam Expressing Breast Cancer Ctcs Using A High Throughput Microfluidic Device, Brandy Charon Snowden
Selection And Enumeration Of Low Epcam Expressing Breast Cancer Ctcs Using A High Throughput Microfluidic Device, Brandy Charon Snowden
LSU Master's Theses
Breast cancer cells overexpress EpCAM that can be used as a marker to select breast cancer CTCs from heterogeneous clinical samples, even when present in low abundance. This study used MDA-MB-231 cells which have a low expression level of EpCAM (1.7 x 103 binding sites per cell.3) Herein, we will report on the use of antiEpCAM antibodies immobilized onto the surface of a capture bed poised within a PMMA microchip that was fabricated into an HTMSU used for selection of breast cancer CTCs seeded into a buffer solution. The microchannel walls within the capture bed were UV modified for 5, …
Integrated Modular Microfluidic System For Forensic Alu Dna Typing, Samuel Kimani Njoroge
Integrated Modular Microfluidic System For Forensic Alu Dna Typing, Samuel Kimani Njoroge
LSU Doctoral Dissertations
Driven by the numerous applications of genome-related research, fully integrated microfluidic systems have been developed that have advanced the capabilities of molecular and, in particular, genetic analyses. A brief overview on integrated microfluidic systems for DNA analysis is given in Chapter 1 followed by a report on micro-capillary electrophoresis (µCE) of Alu elements with laser-induced fluorescence (LIF) detection, in which the monomorphic Alu insertions on the X and Y chromosomes were utilized to detect male DNA in large female DNA background (Y: X = 1:19) without cell sorting prior to the determination. The polymorphic Alu loci with known restricted geographical …
Highly Efficient Selection, Enumeration, Enrichment, And Molecular Profiling Of Low-Abundance Biological Cells, Udara R. Dharmasiri Rasika Dharmasiri
Highly Efficient Selection, Enumeration, Enrichment, And Molecular Profiling Of Low-Abundance Biological Cells, Udara R. Dharmasiri Rasika Dharmasiri
LSU Doctoral Dissertations
After brief overviews of low-abundance cell selection techniques in chapter 1 and circulating tumor cells in chapter 2, this dissertation initially focuses on the development of aptamer incorporated high-throughput microfluidic techniques to select rare circulation prostate cancer cells (LNCaP) directly from whole blood with subsequent quantification of these rare cells using a non-labeling approach. Then, I extended the technology to environmental samples in an effort around time, sensitivity, and portability of traditional groundwater assessment. As a model bio- pathogen, E. coli O157:H7 was chosen due to its toxicity and its adverse impact on recreational waters. Low-abundance (<100 cells mL-1) E. coli O157:H7 cells were isolated and enriched from environmental water samples using a microfluidic chip that its capture beds were covalently decorated with E.coli O157:H7 specific polyclonal antibodies. The selected cells were enumerated using RT-qPCR technique. Finally, I have integrated HTMSU with electrokinetic enrichment microfluidic unit for performance of single recombinant low-abundance CTC cell-based assay. A series of analytical processes were carried out, including immunoaffinity selection of rare CTCs, quantification of selected cells via conductivity impedance and electrophoretic enrichment of selected cells for PCR/LDR/CE interrogation for detection of low-abundance point mutations in genomic DNA.
Microfluidic Devices Interfaced To Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry For Proteomics, Jeonghoon Lee
Microfluidic Devices Interfaced To Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry For Proteomics, Jeonghoon Lee
LSU Doctoral Dissertations
Microfluidic interfaces were developed for off-line matrix-assisted laser desorption/ionization mass spectrometry (MALDI). Microfluidic interfaces allow samples to be manipulated on-chip and deposited onto a MALDI target plate for analysis. For this research, microfluidic culturing devices and automated digestion and deposition microfluidic chip platforms were developed for the identification of proteins. The microfluidic chip components were fabricated on a poly(methyl methacrylate), PMMA, wafer using the hot embossing method and a molding tool with structures prepared via micromilling. One of the most important components of the chip system was a trypsin microreactor. An open channel microreactor was constructed in a 100 µm …
Polymer-Based Microfluidic Devices For High Throughput Single Molecule Detection: Applications In Biological And Drug Discovery, Paul Ichide Okagbare
Polymer-Based Microfluidic Devices For High Throughput Single Molecule Detection: Applications In Biological And Drug Discovery, Paul Ichide Okagbare
LSU Doctoral Dissertations
The realization of high throughput sample processing has become a primary ambition in many research applications with an example being high throughput screening (HTS), which represents the first step in the drug discovery pipeline. Microfluidics is a viable platform for parallel processing of biochemical reactions to increase data production rates due to its ability to generate fluidic networks with a high number of processors over small footprints suitable for optical imaging. Single-molecule detection (SMD) is another technology which has emerged to facilitate the realization of high throughput data processing afforded by its ability to eliminate sample processing steps and generate …
Revolutionizing Genomic Analyses: Mutation Analyses Using Novel Enzyme-Based Assays With Laser-Induced Fluorescence And Polymeric Microfluidic Devices As Electrophoretic Platforms, Rondedrick Deshaun Sinville
Revolutionizing Genomic Analyses: Mutation Analyses Using Novel Enzyme-Based Assays With Laser-Induced Fluorescence And Polymeric Microfluidic Devices As Electrophoretic Platforms, Rondedrick Deshaun Sinville
LSU Doctoral Dissertations
Polymer-based microelectrophoresis was investigated to analyze known (mutation detection) and unknown (mutation scanning) low-abundant mutations in genomic DNA with high diagnostic value for colorectal cancers. For our mutation detection assays, point mutations in the K-ras oncogene were identified using the ligase detection reaction (LDR). For the mutation scanning assay, which searches for sporadic mutations, an EndoV-LDR assay was utilized with mutations in the p53 tumor suppressor gene used as a model. A poly(methylmethacrylate), PMMA, microchip filled with a 4% linear polyacrylamide (LPA) gel was used to electrophoretically sort products formed from LDRs, which produced oligonucleotides <65 bp in length. Using microchip electrophoresis with the LPA, a 44 bp ligation product was resolved from a 100-fold molar excess of unligated primers (25 bp) in approximately 120 s, which was ~17 times faster than conventional capillary gel electrophoresis. In order to simplify the electrophoretic process and further reduce development time, the LDR products were sorted in the absence of the sieving gel using free solution conjugate electrophoresis (FSCE). FSCE incorporated polyamide “drag-tags” onto LDR primers, which provided DNA fragment mobilities in free solution that were dependent upon their size. LDR/drag-tagged (LDR-dt) products could be formed in a multiplexed format for mutant-to-wild-type ratios as low as 1 to 100 with single base resolution. Separations were conducted using capillary array electrophoresis (CAE) and PMMA microchips filled with only a TRIS buffer. Analysis times for the LDR-dt products were less than 11 min using CAE and ~85 s for PMMA microchips with high reproducible migration times within and between microchips. PMMA-based microchips were also evaluated for the identification of sporadic mutations using an endonuclease V – LDR (Endo V/LDR) technique. Endo V cleaves heteroduplexed DNA one base 3’ of single-base mismatched sites as well as nicking DNA at some matched sites as LDR reseals miscleaved sites to reduce false positive signals. Results suggested that Endo V/LDR products from p53 mutations could be successfully separated and detected using a PMMA microfluidic chip filled with a sparsely cross-linked replaceable polyacrylamide gel in less than 6 min, which was approximately 10-fold shorter compared to CAE.
Fabrication Of Dna Microarrays On Poly (Methylmethacrylate) Substrates For Biomolecular Reporting, Catherine N. Situma
Fabrication Of Dna Microarrays On Poly (Methylmethacrylate) Substrates For Biomolecular Reporting, Catherine N. Situma
LSU Doctoral Dissertations
DNA microarrays require the use of substrates with well-established surface modification / probe attachment chemistries. Glass/quartz have been widely adopted as typical support materials since their surface modification chemistries which involve the use of siloxane –based chemistries have been widely studied however; these chemistry is susceptible to hydrolytic cleavage especially at high or low pH values. Recently, polymers have been sought as alternative microarray support materials but their surface modification strategies are not well characterized compared to glass. This report will entail surface photo-modification of PMMA polymer substrates by UV irradiation which produces functional scaffolds of carboxylic groups that allow …
Integrating Micro-Scale Separations To Matrix Assisted Laser Desorption And Ioniation Time Of Flight Mass Spectrometry (Maldi-Tof-Ms) For Protein Analysis, Harrison K. Musyimi
Integrating Micro-Scale Separations To Matrix Assisted Laser Desorption And Ioniation Time Of Flight Mass Spectrometry (Maldi-Tof-Ms) For Protein Analysis, Harrison K. Musyimi
LSU Doctoral Dissertations
This dissertation describes the integration of micro-scale separations to matrix assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI TOF MS) for protein analysis. MALDI MS provides unsurpassed accurate mass measurements of intact bio-molecules, for example peptides and proteins, which in turn generate high molecular specificity enabling the identity, function and structure of these molecules to be characterized. However, in order to realize the full potential of MS in proteomic studies, integrated sample processing on automated and high throughput platforms is required to address the complexity, diversity and the dynamic range of proteomic analysis. The work described here contributes towards …
Two-Dimensional And High-Throughput Electrophoretic Separation Of Proteins Using Polymeric Microchips, Hamed Shadpour
Two-Dimensional And High-Throughput Electrophoretic Separation Of Proteins Using Polymeric Microchips, Hamed Shadpour
LSU Doctoral Dissertations
A major task in proteomics is to identify proteins from a biological sample using two-dimensional (2-D) separation prior to mass spectrometry of peptides generated via proteolytic digestion of the proteins. For 2-D separations, microfluidic devices are superior to bench top and capillary-based systems since they potentially provide higher separation efficiencies due to the minimal dead volumes produced during peak transfer between the two separation dimensions. In addition, fast separations can be envisioned because the column lengths are typically shorter in microfluidic platforms without scarifying peak capacity. High-throughput capabilities are extremely desirable for many types of bio-analytical analyses, such as understanding …
Fabricating Microfluidic Devices In Polymers For Bioanalytica Applications, Sean M. Ford
Fabricating Microfluidic Devices In Polymers For Bioanalytica Applications, Sean M. Ford
LSU Doctoral Dissertations
The research presented in this document focuses on the fabrication, characterization and application of microfluidic systems fabricated in poly(methyl methacrylate) (PMMA) with the emphasis focused on the fabrication processing steps. Microfluidic devices were produced in PMMA using X-ray lithography. The fabrication methods investigated were sacrificial mask, polyimide membrane mask and embossing techniques. PMMA microfluidic devices fabricated using X-ray lithography were characterized using scanning electron microscopy (SEM) and optical microscopy, while analytical techniques such as electroosmotic flow determination, separations, and fluorescent microscopy were used to characterize fluid transport in these devices. A novel method for the heat annealing of PMMA to …