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ItemDesign and Evaluation of the Flexible PDMS Microfluidic Diaphragm Pump with Check-Valve(Georgia Institute of Technology, 2017-05) Kim, SooMicrofluidic pumps in the field of medicine have several applications, including cell separation, microanalysis array, and drug delivery. Among these applications, the pump’s usage for drug delivery requires flow-in-one-direction. Previous attempts for this used standardized check valve from retail store, but this creates complexity in manufacturing and inconsistency of data acquiring due to the failure of manufacture. This thesis is focused on the design and fabrication of flexible micro check valve that can integrate with PDMS microfluidic pump. Both micro check valve and microfluidic channel were fabricated using replica molding, machining, and standard PDMS mixtures. Then, integrated micro pump was studied with compression test to see the performance relative to the result from pump with standardized check valve. Results indicates that integration of PDMS check valve and microfluidic channel is as effective as the one from previous research but there is room for improvement on overall design for integration. The future successful completion of the integration will serve crucial part of this project and can be further developed for drug delivery
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ItemImproving Separation of Differentiated Embryonic Stem Cells with a Microfluidic Device(Georgia Institute of Technology, 2017-05) Gura, Jeremy RDifferences in cell cytoskeletal stiffness can be utilized to sort differentiated embryonic stem cells into distinct populations through the use of a microfluidic device. An initial microfluidic system was developed and proven by previous researchers to sort cells1. Modifications to this microfluidic system were made and aspects have been improved to increase the efficiency of moving large numbers of cells through the device for use in PCR. Preliminary data from the updated microfluidic system shows that vertical integration of cells and increasing cell count along with increasing length of experiment show the greatest promise moving forward. Polymerase Chain Reaction (PCR) is a process to analyze differences in gene expression for genes which produce proteins which may have an effect on cell stiffness. Once total cell throughput is improved to large enough numbers, PCR was then completed in a separate project on the two populations differences in levels of gene expression were compared. The genes to be tested are VIM2, ACTN13, and LMNA4, along with GapDH as a constant, which previous research suggests produce proteins which may play a role in cell stiffness. These genes would therefore have different levels of expression, as measured by PCR, in cells with different levels of cytoskeletal stiffness. Improving the microfluidic separation system will also allow for future use in research, and for commercial use in the field of artificial organ generation, by collecting larger populations of pure populations of stem cells. A system was developed to generate large quantities of cells for the graduate student advisors’ other research endeavors along with other graduate students working on similar projects. Knowing the genes that alter cytoskeletal stiffness will allow for numerous avenues of opportunity, but will greatly change the way populations of cells are isolated and purified.
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ItemMicrofluidic Chip Development and Testing(Georgia Institute of Technology, 2016-12) Camarena, GuillermoThe placenta represents one of biology’s most important membrane’s, yet the study of the characteristics of said membrane is very difficult to simulate in the lab. While there have been methods of creating microfluidic chips to test the biomechanics of this membrane, they require complicated and expensive manufacturing processes. The microfluidic device described in this paper was created with the intent of testing the biomechanic characteristics of the amnion membrane with the use of materials and methods more commonly found in laboratories. The results show that although further testing is required, the microfluidic chip was successful in terms of creating a flow that could be used to test the characteristics of the amnion membrane ex-vivo.
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ItemIdentification of leukemia and cancerous leukocytes based on biophysical properties(Georgia Institute of Technology, 2016-05) Turbyfield, CoryThe goal of the research described in this thesis is to determine size, elasticity, and viscoelasticity of 4 leukocyte cell types, neutrophils, lymphocytes, HL60, and Jurkat. HL60 is a representative of Acute Myeloid Leukemia (AML), and Jurkat is representative of Acute T cell leukemia (ALL). Neutrophils and Lymphocytes are two different types of healthy white blood cells. We believe it is possible to differentiate cells only using mechanical characteristics.
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ItemMulti-Variant Differentiation of Healthy and Cancerous White Blood Cells to Progress the Diagnosis of Leukemia(Georgia Institute of Technology, 2016-05) Crawford, KatherineEvery three minutes, someone in the United States is diagnosed with blood cancer, the most common of which being Leukemia, or the cancer of white blood cells [1]. Distinguishing between healthy white blood cells and leukemia cells has proven to be difficult because the physical and visual similarities between the two. Unlike tumorous cancers, leukemia cells are much more difficult to distinguish and characterize in a clinical setting because of the physiologic nature of the disease. Current diagnostic methods like flow cytometry are known to be time-consuming and costly, creating a crucial need to more effectively identify white blood cells (WBCs) from leukemic cells. The purpose of this study is to use biomechanical markers to characterize the distinct properties of healthy WBC’s and the different types of leukemia. By measuring multiple biomechanical characteristics of each type of cell, each cell type will be able to be narrowed down into a cluster that is representative of the biomechanical characteristics distinctive to that cell type. As we know, it is difficult to distinguish differences in leukemic and healthy WBC’s by a factor of size alone, but adding additional biophysical parameters may lead to improved identification of the pathological condition. Our proposal intends on using four distinct biomechanical parameters to complete our analysis: size, elastic modulus, and both slow and quick viscoelastic response time constants, tau1 and tau2. It has been shown that distinct differences in these specific characteristics do exist between healthy and cancerous WBC’s [2,3], so by analyzing these parameters against each other, we hope to gain a more complete understanding of each of these cell types’ specific biomechanical blueprint for applications in clinical diagnostics.
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ItemMicrofluidic Stiffness-Dependent Separation of Red Blood Cells for Early Malaria Diagnosis and Surveillance(Georgia Institute of Technology, 2013-12-13) Byler, RebeccaThe characterization of the mechanical properties of cells has many broad applications since cell elasticity can indicate pathological state. Notably, many diseases cause significant changes in mechanical properties; for example, at the onset of a malaria infection, the invading parasites can strongly affect the elasticity of Red Blood Cells (RBCs) by causing structural changes. Thus, given the difference in mechanical properties between healthy RBCs and infected RBCs (iRBCs), there exists the potential to separate human blood through microfluidics in order to better detect malaria. We report a statistical difference in cell elasticity between RBCs and chemically mimicked iRBCs, which mimic the pathophysiology of malaria infection, through the use of Atomic Force Microscopy. We demonstrate stiffness-dependent separation of RBCs and chemically mimicked iRBCs via microfluidic technology. The successful completion of this technique will directly aid the long-term objective of this project, which is to develop point-of-care microfluidic technologies for malaria diagnosis and population surveillance that improves on the sensitivity of the existing malaria tests.