Developing Single-Laser Sources for Multimodal Coherent Anti-Stokes Raman Scattering Microscopy

Coherent anti-Stokes Raman scattering (CARS) microscopy has developed rapidly and is opening the door to new types of experiments. This work describes the development of new laser sources for CARS microscopy and their use for different applications. It is specifically focused on multimodal nonlinear optical microscopy—the simultaneous combination of different imaging techniques. This allows us to address a diverse range of applications, such as the study of biomaterials, fluid inclusions, atherosclerosis, hepatitis C infection in cells, and ice formation in cells. For these applications new laser sources are developed that allow for practical multimodal imaging. For example, it is shown that using a single Ti:sapphire oscillator with a photonic crystal fiber, it is possible to develop a versatile multimodal imaging system using optimally chirped laser pulses. This system can perform simultaneous two photon excited fluorescence, second harmonic generation, and CARS microscopy. The versatility of the system is further demonstrated by showing that it is possible to probe different Raman modes using CARS microscopy simply by changing a time delay between the excitation beams. Using optimally chirped pulses also enables further simplification of the laser system required by using a single fiber laser combined with nonlinear optical fibers to perform effective multimodal imaging. While these sources are useful for practical multimodal imaging, it is believed that for further improvements in CARS microscopy sensitivity, new excitation schemes are necessary. This has led to the design of a new, high power, extended cavity oscillator that should be capable of implementing new excitation schemes for CARS microscopy as well as other techniques. Our interest in multimodal imaging has led us to other areas of research as well. For example, a fiber-coupling scheme for signal collection in the forward direction is demonstrated that allows for fluorescence lifetime imaging without significant temporal distortion. Also highlighted is an imaging artifact that is unique to CARS microscopy that can alter image interpretation, especially when using multimodal imaging. By combining expertise in nonlinear optics, laser development, fiber optics, and microscopy, we have developed systems and techniques that will be of benefit for multimodal CARS microscopy.

Long-term hydroclimatic change and interannual variability in water sources, Apex River (Iqaluit), Baffin Island, Nunavut

The eastern Canadian Arctic is home to Canada’s largest Indigenous population, which depends on local freshwater sources for drinking water. However, small watersheds have rarely been analyzed for long-term hydrologic response to changing climate. This study aims to address this issue by examining the Apex River, a small watershed with a long hydroclimatic record, near Iqaluit, Nunavut. Particular emphasis was placed on the long-term changes in climate and river discharge, and the seasonal variability of water sources between two snapshots in time, 1983 and 2013. Long-term hydrological data were obtained from gauge station 10UH002, operated by Environment and Climate Change Canada, and long-term meteorological data were acquired from Environment Canada–operated stations near Iqaluit Airport. Breakpoint analysis suggested that long-term mean annual surface air temperatures have increased since 1994. In contrast, no long-term total precipitation or annual discharge changes were observed. However, river flow initiation and cessation analyses of the Apex River flow season indicates that flow extended into the autumn since the 2000s. The 2013 flow season lasted 44 days longer than the 1983 flow season. Systematic river sampling was undertaken throughout the 2013 thaw season to determine contributing proportions of event (snowmelt or rainfall) and pre-event (baseflow) water to river runoff. Results from the stable isotope hydrograph separation for 2013 were compared to findings for 1983. Snow was the main source of water to the river during the snowmelt period in 1983 and 2013, however baseflow was still an important contributor. Although there was high similarity of water sources early in the season in 1983 and 2013, the two years differed during the autumn. In 2013 there was a high rainfall runoff response that was not present in 1983, suggesting high release of late-season sub-surface water storage and an increased sensitivity to late-season rainfall events in 2013. This research provides insights into the hydrologic response of the Apex River to long-term climatic change, and highlights the need for high-quality precipitation and discharge data for effective long-term hydrological assessment.

ROCK OUTCROPS IN THE CANADIAN SHIELD: AN INVESTIGATION OF CONTAMINANT TRANSPORT FROM SURFACE SOURCES IN FRACTURED ROCK AQUIFERS

Flow, recharge and transport dynamics in fractured rock aquifers with low lying rock outcrops is a largely unexplored area of study in hydrogeology. The purpose of this thesis is to examine these topics in an agricultural area in Eastern Ontario. The study consists of a regional scale groundwater quality study, an infiltration experiment that considers bacteria transport from the ground surface to a well, and a numerical modelling study that tests the parameters that affect surface infiltration of a tracer from a rock outcrop to a deeper horizontal fracture. In the water quality study, approximately 65% of the samples contained total coliform, 16% contained E. coli, and 1% contained nitrate-N at greater than 5 mg/L. Occurrence of E. coli increased when considering seasonality, where wells were drilled on rock outcrops, and for shallow well intervals. Nitrate-N did not occur above the Guidelines for Canadian Drinking Water Quality (Health Canada, 2012) of 10 mg/L. Rapid arrival times were observed in the infiltration study for both the microspheres (30 minutes) and a dye tracer (45 minutes) in a well approximately 6.0 m in horizontal and 2.8 m in vertical distance from the tracer source. Transport velocities were approximately 38.9 m/day for the dye tracer and 115.2 m/day for the colloidal tracer. Results of the model runs indicate that overburden can provide an effective protective layer to transport in fractures, that high groundwater velocities occur in larger fracture apertures and higher gradients dilute tracer concentrations, and that lower groundwater velocities occur with smaller fracture apertures and lower gradients result in elevated tracer concentrations. Lower rainfall rates, larger fracture apertures, early tracer time, larger gradients, and lower water levels maintained unsaturated conditions for longer time periods such that tracer transport was delayed until saturated conditions were attained. The overall heterogeneity of this aquifer environment creates a source water protection conundrum where the water quality is generally good, while transport can occur very quickly in proximity to rock outcrops and in areas with limited overburden.

#DressCodePM & Re-Framing The Niqab: News Sources, Hashtag Activism, and Media Representation

This thesis originates from my interest in exploring how minorities are using social media to talk back to mainstream media. This study examines whether hashtags that trend on Twitter may impact how news stories related to minorities are covered in Canadian media. The Canadian Prime Minister Stephen Harper stated the niqab was “rooted in a culture that is anti-women” on 10 March 2015. The next day #DressCodePM trended in response to the PM’s niqab remarks. Using network gatekeeping theory, this study examines the types of sources quoted in the media stories published on 10 and 11 March 2015. The study’s goal is to explore whether using tweet quotes leads to the representation of a more diverse range of news sources. The study compares the types of sources quoted in stories that covered Harper’s comments without mentioning #DressCodePM versus stories that mention #DressCodePM. This study also uses Tuen A. van Dijk’s methodology of asking “who is speaking, how often and how prominently?” in order to examine whose voices have been privileged and whose voices have been marginalized in covering the niqab in Canadian media from the 1970s and until the days following the PM’s remarks. Network gatekeeping theory is applied in this study to assess whether the gated gained more power after #DressCodePM trended. The case study’s findings indicates that Caucasian male politicians were predominantly used as news sources in covering stories related to the niqab for the past 38 years in the Globe and Mail. The sourcing pattern of favouring politicians continued in Canadian print and online media on 10 March 2015 following Harper’s niqab comments. However, ordinary Canadian women, including Muslim women, were used more often than politicians as news sources in the stories about #DressCodePM that were published on 11 March 2015. The gated media users were able to gain power and attract Canadian Media’s attention by widely spreading #DressCodePM. This study draws attention to the lack of diversity of sources used in Canadian political news stories, yet this study also shows it is possible for the gated media users to amplify their voices through hashtag activism.

The relationship of E. coli and total coliform culture to the quantitative molecular detection of various fecal indicators, sources, and pathogens in private drinking water wells

Water remains a predominant vector for human enteric pathogens not just for developing countries but also developed nations, where numerous infectious disease outbreaks, linked to the contamination of drinking water have been documented. Private drinking water wells are a source of drinking water that is largely unstudied even though a significant percentage of the population in Ontario relies on wells as their primary water source. As there exists little to no systematic surveillance for enteric infections or outbreaks related to well water sources, these individuals may be at higher risk of waterborne infectious diseases. The relationships between various fecal indicators in the water of private drinking water wells, including E. coli, Total Coliforms (TC) and Bacteroides, and enteric pathogens, including Campylobacter jejuni, Salmonella spp., and Shiga toxin producing E. coli, were studied. Convenience private well water samples collected from various regions of interest during the summer of 2014 underwent membrane filtration and culture to determine quantities of E. coli and TC colony forming units. 289 E. coli positive and 230 TC-only waters were successfully analyzed by individual qPCR assays for the aforementioned enteric pathogens. Microbial source tracking methods targeted to specific Bacteroides were used to determine the source of fecal contamination as either human or bovine. The source of fecal contamination varied by geographic region and is thought to be due to such things as differences in septic tank density and underlying geology, among others. Fecal indicators, E. coli and Bacteroides, were significantly correlated. E. coli as measured by qPCR was more strongly correlated to both total and human-specific Bacteroides genetic markers than culturable E. coli. Lastly, 1.9% of samples showed molecular evidence of contamination with enteric pathogens. Although low, this finding is significant given the limited volume of water available for testing, and suggests a potential health risk to consumers. Knowing the extent of contamination, as well as the biologic source, can better inform risk assessment and the development of potential intervention strategies for private well water in specific regions of Ontario.

RED COATS AND WILD BIRDS: MILITARY CULTURE AND ORNITHOLOGY ACROSS THE NINETEENTH-CENTURY BRITISH EMPIRE

“Red coats and wild birds: military culture and ornithology across the nineteenth-century British Empire” investigates the intersections between British military culture and the practices and ideas of ornithology, with a particular focus on the British Mediterranean. Considering that British officers often occupied several imperial sites over the course of their military careers, to what extent did their movements shape their ornithological knowledge and identities at “home” and abroad? How did British military naturalists perceive different local cultures (with different attitudes to hunting, birds, field science, etc.) and different local natures (different sets of birds and environments)? How can trans-imperial careers be written using not only textual sources (for example, biographies and personal correspondence) but also traces of material culture? In answering these questions, I centre my work on the Mediterranean region as a “colonial sea” in the production of hybrid identities and cultural practices, and the mingling of people, ideas, commodities, and migratory birds. I focus on the life geographies of four military officers: Thomas Wright Blakiston, Andrew Leith Adams, L. Howard Lloyd Irby, and Philip Savile Grey Reid. By the mid-nineteenth century, the Mediterranean region emerged as a crucial site for the security of the British “empire route” to India and South Asia, especially with the opening of the Suez Canal in 1869. Military stations served as trans-imperial sites, connecting Britain to India through the flow of military manpower, commodities, information, and bodily experiences across the empire. By using a “critical historical geopolitics of empire” to examine the material remnants of the “avian imperial archive,” I demonstrate how the practices and performances of British military field ornithology helped to: materialize the British Mediterranean as a moral “semi-tropical” place for the physical and cultural acclimatization of British officers en route to and from India; reinforce imperial presence in the region; and make “visible in new ways” the connectivity of North Africa to Europe through the geographical distribution of birds. I also highlight the ways in which the production of ornithological knowledge by army officers was entwined with forms of temperate martial masculinity.

A Study of the Development of the Ability of the Archives Department of the Hudson's Bay Company to Carry Out Document Repairs

The objective of this thesis was to determine whether the establishment and operation of an archives services by the Hudson's Bay Company had an effect on the company's ability to carry out document repairs. Data collection methods included reviews of published material, archival records of the Hudson's Bay Company, and semi-structured interviews. The study found that the Hudson's Bay Company's commitment to operating a modern archives service in accordance with accepted archive administration practices had a substantial effect on its ability to carry out document repairs. The principled approach to repair, as practiced by the Public Record Office, was a major influence. A review of secondary sources placed this development squarely within the context of archival developments in 20th century England. Overall, the thesis findings add to the growing conversation about conservation history in England, in particular, archive conservation history as it occurred outside of the Public Record Office in the 20th century, by discussing how some methods of repair that were devised, adopted and extended by the Public Record Office in the 19th and 20th centuries were adopted and applied in the 20th century by a well-established business corporation.

Glycemic State Regulates Brain-Derived Neurotrophic Factor Responsiveness of Neurons in the Paraventricular Nucleus of the Hypothalamus

Brain derived neurotrophic factor (BDNF) is a member of the family of neurotrophins and binds to the tropomyosin-related kinase B (TrkB) receptor. Like other neurotrophic factors, BDNF is involved in the development and differentiation of neurons. Recently, studies have suggested important roles for BDNF in the regulation of energy homeostasis. The paraventricular nucleus (PVN) is critical for normal energy balance contains high levels of both BDNF and TrkB mRNA. Studies have shown that microinjections of BDNF into the PVN increase energy expenditure, suggesting BDNF plays a role in energy homeostasis through direct actions in this hypothalamic nucleus. We used male Sprague-Dawley rats to perform whole-cell current-clamp experiments from PVN neurons in slice preparation. BDNF was bath applied at a concentration of 2nM and caused depolarizations in 54% of neurons (n = 25; mean change in membrane potential: 8.9 ± 1.2 mV), hyperpolarizations in 23% (n = 11; mean change in membrane potential: -6.7 ± 1.4 mV), while the remaining cells tested were unaffected. Previous studies showing effects of BDNF on γ-aminobutyric acid type A (GABAA) mediated neurotransmission in PVN led us to examine if these BDNF-mediated changes in membrane potential were maintained in the presence of tetrodotoxin (TTX) sodium channel blocker (N = 9; 56% depolarized, 22% hyperpolarized, 22% non-responders) and bicuculline (GABAA antagonist) (N = 12; 42% depolarized, 17% hyperpolarized, 41% non-responders), supporting the conclusion that these effects on membrane potential were postsynaptic. We also evaluated the effects of BDNF on these neurons across varying physiologically relevant extracellular glucose concentrations. At 10 mM 23% (n = 11; mean: -6.7 ± 1.4 mV) of PVN neurons hyperpolarized in response to BDNF treatment, whereas at 0.2 mM glucose, 71% showed hyperpolarizing effects (n = 12; mean: -6.3 ± 2.8 mV). Our findings reveal that BDNF has direct impacts on PVN neurons and that these neurons are capable of integrating multiple sources of metabolically relevant input. Our analysis regarding glucose concentrations and their effects on these neurons’ response to other metabolic signals emphasizes the importance of using physiologically relevant conditions for study of central pathways involved in the regulation of energy homeostasis.