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You searched for subject:(biomolecular fluorescence detection). Showing records 1 – 2 of 2 total matches.

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Penn State University

1. Kumar, Nitin. ENGINEERING NOVEL SUBSTRATES FOR ADVANCED BIOMOLECULAR DETECTION.

Degree: PhD, Chemical Engineering, 2008, Penn State University

Detection of biological species is central to many areas of biology and life sciences such as ultra sensitive disease detection, biochemical sensing and targeted drug delivery. Currently, detection of biomolecules like proteins and DNA is done by fluorescence detection, however, enhancing detection sensitivity and increasing signal to noise ratio still remains a major challenge. Novel techniques are presently required which can provide reliable, low cost, and ultra-sensitive detection of proteins and DNA. This thesis will focus on two approaches in this context: the use of diblock copolymer templates and zinc oxide nanorods, with an aim to provide rapid, sensitive and accurate biomolecular detection. While complementary, these approaches are not necessarily interlinked. First approach will focus on utilizing the microphase separation behavior of diblock copolymers to pattern proteins with nanometer periodicity. The structural variety and chemical heterogeneity of polystyrene-block-poly(methylmethacrylate) (PS-b-PMMA) and polystyrene-b-poly(4-vinylpyridine) (PS-b-PVP) template surfaces were successfully exploited to spontaneous formation of self-assembled, linear and hexagonally-ordered protein arrays that exhibit repeat spacings in a nanoscopic dimension. More importantly, protein molecules on the polymeric templates maintained their natural conformation and activity for several months. Our results demonstrate that self-assembling, chemically heterogeneous, diblock copolymer templates can be used as excellent, high payload, high density protein templates making them highly suitable as functional substrates in many proteomics applications. Second approach will focus on the remarkably enhanced optical detection of DNA and proteins which is enabled by the use of nanoscale zinc oxide platforms. Using model protein and nucleic acid systems, we demonstrate that engineered nanoscale zinc oxide nanostructures can significantly enhance the detection capability of biomolecular fluorescence. Without any chemical or biological amplification processes, nanoscale zinc oxide platforms enabled increased fluorescence detection of these biomolecules when compared to other commonly used substrates such as glass, quartz, polymer, and silicon. We also demonstrate the easy integration potential of zinc oxide nanostructures into periodically patterned platforms which, in turn, will promote the assembly and fabrication of these materials into multiplexed, high-throughput, optical sensor arrays.

Subjects/Keywords: biomolecular fluorescence detection; diblock copolymers; protein arrays; zinc oxide nanorods

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Kumar, N. (2008). ENGINEERING NOVEL SUBSTRATES FOR ADVANCED BIOMOLECULAR DETECTION. (Doctoral Dissertation). Penn State University. Retrieved from https://etda.libraries.psu.edu/catalog/8553

Chicago Manual of Style (16th Edition):

Kumar, Nitin. “ENGINEERING NOVEL SUBSTRATES FOR ADVANCED BIOMOLECULAR DETECTION.” 2008. Doctoral Dissertation, Penn State University. Accessed January 20, 2020. https://etda.libraries.psu.edu/catalog/8553.

MLA Handbook (7th Edition):

Kumar, Nitin. “ENGINEERING NOVEL SUBSTRATES FOR ADVANCED BIOMOLECULAR DETECTION.” 2008. Web. 20 Jan 2020.

Vancouver:

Kumar N. ENGINEERING NOVEL SUBSTRATES FOR ADVANCED BIOMOLECULAR DETECTION. [Internet] [Doctoral dissertation]. Penn State University; 2008. [cited 2020 Jan 20]. Available from: https://etda.libraries.psu.edu/catalog/8553.

Council of Science Editors:

Kumar N. ENGINEERING NOVEL SUBSTRATES FOR ADVANCED BIOMOLECULAR DETECTION. [Doctoral Dissertation]. Penn State University; 2008. Available from: https://etda.libraries.psu.edu/catalog/8553

2. Liu, Cong, Ph. D. Single-molecule tracking and its application in biomolecular binding detection.

Degree: PhD, Biomedical Engineering, 2017, University of Texas – Austin

In the past two decades significant advances have been made in single-molecule detection, which enables the direct observation of single biomolecules at work in real time under physiological conditions. In particular, the development of single-molecule tracking (SMT) microscopy allows us to monitor the motion paths of individual biomolecules in living systems, unveiling the localization dynamics and transport modalities of the biomolecules that support the development of life. While 3D-SMT is probably the most suitable method for determining whether tracked molecules (can be any biomolecule such as DNA, membrane receptors, and transcription factors) form dimers or complexes with other molecules, great technical challenges remain to be overcome before the potential of 3D-SMT in biomolecular binding detection can be realized. This dissertation describes my work on recent methodology development to overcome these challenges, and new applications of the 3D-SMT technology in rare molecular species quantification. First, we provide an overview of current SMT technologies, with an emphasis on three-dimensional feedback controlled SMT. Advantages and drawbacks of each SMT method are outlined. Second, we describe the theoretical modeling and instrumentation of our confocal tracking microscope. Its multi-dimensional sensing capability (3D position, diffusion coefficient, fluorescence lifetime) is experimentally characterized. In order to maximize the tracking duration, we have also developed strategies to effectively slow-down fast diffusing molecule, and optimized the buffer conditions. Third, we show that our 3D-SMT microscope can detect biomolecular association/disassociated by two types of contrast mechanisms: diffusion rate and lifetime FRET signal. DNA transient binding is used as a model system because of ease of fluorescent labeling and tunable binding kinetics. Both of the two mechanisms involve tracking a fluorescent-labeled single-stranded DNA (ssDNA), but the second approach also requires its complementary strand to be labeled by a dark quencher. A combined analysis of multiple single-molecule trajectories allow us to measure the kinetics that is even beyond the physical bandwidth of the tracking system. In the end, we introduce the application of SMT in rare single-molecule species quantification. The theory for predicting the sensitivity and fidelity is established. Our work highlights the fundamental limitations that we are facing in precise single-molecule identification and quantification without amplification. Advisors/Committee Members: Yeh, Tim H. C. (advisor), Dunn, Andrew K (committee member), Stachowiak, Jeanne C (committee member), Brown, Richard M (committee member), Wang, Yaguo (committee member).

Subjects/Keywords: Single-molecule detection; Fluorescence lifetime; Single-molecule tracking; Biomolecular binding detection; Single biomolecules

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Liu, Cong, P. D. (2017). Single-molecule tracking and its application in biomolecular binding detection. (Doctoral Dissertation). University of Texas – Austin. Retrieved from http://hdl.handle.net/2152/47332

Chicago Manual of Style (16th Edition):

Liu, Cong, Ph D. “Single-molecule tracking and its application in biomolecular binding detection.” 2017. Doctoral Dissertation, University of Texas – Austin. Accessed January 20, 2020. http://hdl.handle.net/2152/47332.

MLA Handbook (7th Edition):

Liu, Cong, Ph D. “Single-molecule tracking and its application in biomolecular binding detection.” 2017. Web. 20 Jan 2020.

Vancouver:

Liu, Cong PD. Single-molecule tracking and its application in biomolecular binding detection. [Internet] [Doctoral dissertation]. University of Texas – Austin; 2017. [cited 2020 Jan 20]. Available from: http://hdl.handle.net/2152/47332.

Council of Science Editors:

Liu, Cong PD. Single-molecule tracking and its application in biomolecular binding detection. [Doctoral Dissertation]. University of Texas – Austin; 2017. Available from: http://hdl.handle.net/2152/47332

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