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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Design and Characterization of a Miniaturized Fluorescence Analysis System for Measurement of Cell-Free DNA

Bondi, Parker 30 November 2018 (has links)
Sepsis is a dysregulated systemic response to infection and is one of the leading causes of in-hospital mortality in Canada. Accurate distinction between survivors and non-survivors of sepsis has recently been demonstrated through quantification of cell-free DNA (cfDNA) concentration in blood. In an analysis of 80 septic patients, non-survivors of sepsis had significantly higher cfDNA concentration levels than that of survivors or healthy patients. Real time separation of cfDNA from contaminants in blood has also been done using a cross channel microfluidic device. Current methods for DNA quantification utilize time consuming and complicated laboratory equipment and therefore are not suitable for bedside real-time testing. Thus a handheld cfDNA fluorescence device coined the Sepsis Check was designed that can perform DNA characterization in a reservoir device and DNA detection in a microfluidic cross channel device. The goal is to use this system along with the cross channel devices to set apart survivors or healthy donors from non-survivors in patients with sepsis. The design consists of a 470𝑛𝑚 light emitting diode (LED) with 170𝑚𝑊 of optical power (LED470L – ThorLabs), an aspherical uncoated lens with a focal length of 15𝑚𝑚 (LA1540-ML – ThorLabs), a 488𝑛𝑚 bandpass filter with a 3𝑛𝑚 full width at half maximum (FWHM) (FL05488-3 – ThorLabs), an aspherical uncoated lens with a focal length of 25𝑚𝑚 (LA1560-ML – ThorLabs), an aspherical uncoated lens with a focal length of 35𝑚𝑚 (LA1027-ML – ThorLabs), a 525𝑛𝑚 longpass filter with an optical density >4.0 (F84744 – Edmund Optics), and a Raspberry Pi Camera V2 (Raspberry Pi Foundation). The Sepsis Check is made to excite the dsDNA specific PicoGreen fluorophore which has a peak absorbance at 502𝑛𝑚 and a peak emission at 523𝑛𝑚. In summary, the Sepsis Check in this thesis is capable of calibrating dsDNA concentration from 1𝜇𝑔/𝑚𝐿 to 10𝜇𝑔/𝑚𝐿 and detect DNA accumulation of 5𝜇𝑔/𝑚𝐿 and 10𝜇𝑔/𝑚𝐿 in the cross channel device. This tool can be a valuable addition to the ICU to rapidly assess the severity of sepsis for informed decision making. / Thesis / Master of Applied Science (MASc)

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