Surface quality plays a vital role in controlling the function performance of the workpiece. With the development of the measuring technique, areal surface measurement has been widely applied in the industry. However, estimating the uncertainty of areal surface measurement is still a challenge. Except for the metrological characteristics of the measurement system, measurement conditions should be considered for uncertainty evaluation. The dissertation investigates the influence of measurement settings on surface measurement. A silver-plated surface, three different rough grinding surfaces, and three different rough cylindrical grinding surfaces were measured using structured illumination microscopy. The measurements were at the different objective lenses, vertical scanning interval, exposure time, and sample tilt. The results show that the measurement settings influence the non-measured points, measurement noise, and areal surface texture parameters. Therefore, according to the investigation, the sample tilt and exposure time should also be included in the uncertainty budget. An approach was proposed to investigate the influence of non-measured points on the areal surface texture parameters. The relation between the non-measured points ratio and measurement settings was investigated, and how the areal surface texture parameters changed due to the non-measured points was studied. Moreover, an approach based on the metrological characteristic method was proposed to estimate the uncertainty due to the measurement noise. This method can be extended to the uncertainty evaluation due to other metrological characteristics. Additionally, an approach based on the Monte Carlo Method was proposed to estimate the measurement uncertainty due to different influences. This approach was verified as feasible in the practical measurement.

Laser application in the field of biological and medical sciences has significantly grown, thereby strengthening the field of Biophotonics. Research conducted in Biophotonics focuses on the concept of using light especially in the visible and near infrared regions of the electromagnetic radiation for the evaluation of living systems. In this thesis new discoveries are presented about low level laser therapy, optical trapping, transmission spectroscopy, luminescence spectroscopy and structured illumination microscopy (SIM), displaying the impact each technique has on HIV infected cells. The results showed that the irradiation of HIV-1 infected TZM-bl cells with low power red laser reduces HIV-1 infection. The outcomes of this study further proved that when irradiation is used in conjunction with efavirenz, an antiretroviral drug, HIV-1 infection could be reduced to undetectable levels in TZM-bl cells. Through the coupling of transmission spectroscopy with optical trapping, and separately, use of luminescence spectroscopy, label free diagnosis of HIV in infected cell samples was achieved. This finding affirms that HIV-1 infection can be detected in a label free manner when using laser based techniques. Furthermore, the photoluminescence spectrometer system was employed to generate a decay curve, which was necessary so as to have some understanding on lifetime of the luminescent signal in infected TZM-bl cells. Finally, in order to confirm that indeed TZM-bl cells were infected, an established super-resolution microscopy system SIM was used to detect HIV-1 infection in TZM-bl cells. Indeed in the infected cells viral molecules p24 and gp41 were detected through SIM, while they were not detected in uninfected cells. In future studies, super resolution microscopy would be coupled to an optical trapping system in order to confirm that each trapped cells is whether infected or uninfected so as to improve HIV diagnosis. / College of Science, Engineering and Technology / Ph. D. (Science, Engineering and Technology)

Manual inspection is a prevailing practice for quality assessment of poultry meat, but it is labor-intensive, tedious, and subjective. This thesis aims to assess the efficacy of an emerging structured illumination reflectance imaging (SIRI) technique with machine learning approaches for assessing WS and microbial spoilage in broiler breast meat. Broiler breast meat samples were imaged by an in house-assembled SIRI platform under sinusoidal illumination. In first experiment, handcrafted texture features were extracted from direct component (DC, corresponding to conventional uniform illumination) and amplitude component (AC, unique to the use of sinusoidal illumination) images retrieved from raw SIRI pattern images build linear discriminant analysis (LDA) models for classifying normal and defective samples. A further validation experiment was performed using deep learning as a feature extractor followed by LDA. The third experiment was on microbial spoilage assessment of broiler meat, deep learning models were used to extract features from DC and AC images builds on classifiers. Overall, this research has demonstrated consistent improvements of AC over DC images in assessing WS and spoilage of broiler meat and that SIRI is a promising tool for poultry meat quality detection.

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Neutrophile sind die häufigsten weißen Blutkörperchen im menschlichen Blut. Sie bilden die erste Verteidigungslinie und töten eindringende Krankheitserreger ab. Neutrophile extrazelluläre Fallen (NETs) sind netzartige Strukturen, die aus dekondensiertem Chromatin bestehen und mit zytotoxischen Proteinen dekoriert sind. NETs können Mikroben in vitro und in vivo einfangen und abtöten, sind aber auch für verschiedene Krankheiten verantwortlich. Frühere Studien haben eine spezifische Gruppe von 20-50 Neutrophilenproteinen identifiziert, die an NETs gebunden sind und von denen einige eine mikrobizide Wirkung haben. Wie diese Proteine an die NETs binden, wie sie interagieren und wie die Bindung ihre antimikrobielle Aktivität beeinflusst, ist noch nicht bekannt. In dieser Dissertation habe ich die Verteilung von acht neutrophilen Proteinen und Nukleosomen auf NETs mit Hilfe der Superauflösungsmikroskopie untersucht. Es wurden drei unabhängige Techniken mit Auflösungen von mehr als 90 nm verwendet. Die Nukleosomen bildeten auf den NETs periodische Cluster mit deutlich größeren Abständen im Vergleich zum kondensierten Chromatin. Drei NET-Proteine waren ebenfalls in periodischen Clustern auf den NETs lokalisiert und zwei von ihnen waren stark mit Nukleosomen kolokalisiert. Alle anderen analysierten Proteine zeigten keine Muster der Bindung an NETs. Zusammengenommen zeigen diese Ergebnisse, dass die Bindung von Proteinen an NETs zumindest teilweise spezifisch ist und teilweise durch Wechselwirkungen mit Nukleosomen vermittelt wird. Die erfolgreiche Einführung der superauflösenden Mikroskopie für schwierige NET-Proben in Kombination mit einem vorgeschlagenen rekonstituierten NET-System eröffnet neue Möglichkeiten für das Verständnis der molekularen Mechanismen der NET-Bildung und der Protein-Protein-Interaktion bei der NET-vermittelten Abtötung. / Neutrophils are the most abundant human white blood cell in circulation. They are the first line of defense and kill invading pathogens. Neutrophil Extracellular Traps (NETs) are weblike structures composed of decondensed chromatin decorated with cytotoxic proteins. NETs can trap and kill microbes in vitro and in vivo, but also mediate several diseases. Previous studies identified a specific set of 20-50 neutrophil proteins bound to NETs, several with microbicidal activity. It remains unknown how these proteins bind to NETs, how they interact and how binding influences their anti-microbial activity. In this dissertation, I studied the distribution of eight neutrophil proteins and nucleosomes on NETs using super-resolution microscopy. Three independent techniques with resolutions larger than 90nm were used. Nucleosomes formed periodic clusters on NETs, with significantly larger spacing compared to condensed chromatin. Three NET proteins also localized in periodic clusters on NETs and two of them strongly co-localized with nucleosomes. All other proteins analyzed showed no patterns binding to NETs. Taken together, these findings demonstrate that, at least some, protein binding to NETs is specific and in part mediated by interactions with nucleosomes. The successful introduction of super-resolution microscopy to the challenging NET samples in combination with a proposed reconstituted NET system opens new possibilities to understand the molecular mechanisms behind NET formation and protein-protein interaction in NET mediated killing.