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Item An inexpensive, versatile, compact, programmable temperature controller and thermocycler for simultaneous analysis and visualization within a microscope(Springer Science and Business Media LLC, 2021-05) Cruz, Pablo Martínez; Wood, Mikayla A.; Abbasi, Reha; LeFevre, Thomas B.; McCalla, Stephanie E.Microfluidic Lab on a Chip (LOC) devices are key enabling technologies for research and industry due to their compact size, which increases the number of integrated operations while decreasing reagent use. Common operations within these devices such as chemical and biological reactions, cell growth, or kinetic measurements often require temperature control. Commercial temperature controllers are constrained by cost, complexity, size, and especially versatility for use in a broad range of applications. Small companies and research groups need temperature control systems that are more accessible, which have a wide applicability. This work describes the fabrication and validation of an inexpensive, modular, compact, and user-friendly temperature control system that functions within a microscope. This system provides precise temperature acquisition and control during imaging of any arbitrary sample which complies with the size of a microscope slide. The system includes two parts. The first part is a compact and washable Device Holder that is fabricated from high temperature resistant material and can fit securely inside a microscope stage. The second part is a robust Control Device that incorporates all the necessary components to program the temperature settings on the device and to output temperature data. The system can achieve heating and cooling times between 50°C and 100°C of 32 seconds and 101 seconds, respectively. A Bluetooth enabled smartphone application has been developed for real-time data visualization. The utility of the temperature control system was shown by monitoring rhodamine B fluorescence in a microfluidic device over a range of temperatures, and by performing a polymerase chain reaction (PCR) within a microscope. This temperature control system could potentially impact a broad scope of applications that require simultaneous imaging and temperature control.Item Coupling fluid flow to hydrogel fluidic devices with reversible “pop-it” connections(Royal Society of Chemistry, 2021-01) Abbasi, Reha; LeFevre, Thomas B.; Benjamin, Aaron D.; Thornton, Isaak J.; Wilking, James N.Here, we describe a simple, reversible, plug-based connector designed to couple microfluidic tubing to a hydrogel-based fluidic device, to allow for pressurized liquid flow through the system.Item Improving transport in hydrogels for 3D bioprinting applications(Montana State University - Bozeman, College of Engineering, 2021) Abbasi, Reha; Chairperson, Graduate Committee: James Wilking; Aaron D. Benjamin was an author and Madison Owens, Robert J. Olsen, Danica J. Walsh, Thomas B. LeFevre and James N. Wilking were co-authors of the article, 'Light-based 3D printing of hydrogels with high-resolution channels' in the journal 'Biomedical physics & engineering express' which is contained within this dissertation.; Thomas B. LeFevre was an author and Aaron D. Benjamin, Isaak J. Thornton, and James N. Wilking were co-authors of the article, 'Coupling fluid flow to hydrogel fluidic devices with reversible "pop-it" connections' in the journal 'Lab on a chip' which is contained within this dissertation.; Zahra Mahdieh was an author and Galip Yiyen, Robert A. Walker and James N. Wilking were co-authors of the article, 'Light-based 3D bioprinting of hydrogels containing colloidal calcium peroxide' submitted to the journal 'Bioprinting' which is contained within this dissertation.Hydrogels are soft, water-based gels with widespread applications in medicine, tissue engineering, and biotechnology. Many of these applications require structuring hydrogels in three-dimensional space. Light-based 3D printers offer exquisite spatial control; however, technologies for light-based 3D-printing of hydrogels remain limited. This is mainly caused by poor material transportation through the hydrogel. For example, limited transport of oxygen and other nutrients through 3D printed tissue constructs containing living cells leads to low cell viability. Here, we describe three experimental research studies focused on improving material transport in 3D-printed hydrogels. In the first part of this thesis, we describe a generalizable method for light-based 3D printing of hydrogels containing open, well-defined, submillimeter-scale channels with any orientation. These submillimeter channels allow for bulk liquid flow through the hydrogel to improve nutrient and oxygen transport. In the second part of this thesis, we describe a simple, reversible, plug-based connector designed to couple tubing to a hydrogel-based fluidic device to allow for pressurized liquid flow through the system. The resulting connection can withstand liquid pressures significantly greater than traditional, connector-free approaches, enabling long-term flow through 3D-printed hydrogels. In the third part of this thesis, we characterize the printability of photopolymerizable resins containing particles that slowly dissolve to release oxygen and thereby improve cell viability. The light-based 3D bioprinting technologies we describe in this thesis will improve material transport through 3D printed hydrogels and enable a wide variety of applications in 3D bioprinting and hydrogel fluidics.Item Light-Based 3D Printing of Hydrogels with High-Resolution Channels(2019-01) Benjamin, Aaron D.; Abbasi, Reha; Owens, Madison; Olsen, Robert J.; Walsh, Danica J.; LeFevre, Thomas B.; Wilking, James N.Hydrogels are soft, water-based gels with widespread applications in personal care products, medicine and biomedical engineering. Many applications require structuring the hydrogel into complex three-dimensional (3D) shapes. For these applications, light-based 3D printing methods offer exquisite control over material structure. However, the use of these methods for structuring hydrogels is underdeveloped. In particular, the ability to print hydrogel objects containing internal voids and channels is limited by the lack of well-characterized formulations that strongly attenuate light and the lack of a theoretical framework for predicting and mitigating channel occlusion. Here we present a combined experimental and theoretical approach for creating well-defined channels with any orientation in hydrogels using light-based 3D printing. This is achieved by the incorporation of photoblocker and the optimization of print conditions to ensure layer-layer adhesion while minimizing channel occlusion. To demonstrate the value of this approach we print hydrogels containing individual spiral channels with centimeter-scale length and submillimeter-scale cross-section. While the channels presented here are relatively simple, this same approach could be used to achieve more complex channel designs mimicking, for example, the complex vasculature of living organisms. The low cytotoxicity of the gel makes the formulation a promising candidate for biological applications.