Among the central constituents of the innate immune system are macrophages, which are known for phagocytosis or ‘eating’ foreign particles or pathogens. Macrophages express several cell-surface proteins including transmembrane and membrane-anchored receptors, which play a vital role in their response to pathogenic stimuli. The plasma membrane is a highly fluid and dynamic environment, which facilitates the diffusion of lipids and proteins within the plane of the membrane. This study aims to measure the lateral diffusion of two types of plasma membrane receptors on macrophages, toll-like receptor II (TLR2) and cluster of differentiation 14 (CD14), to answer three main research questions: 1) Which type of fluorescence-based microscopy techniques is best suited for measuring the lateral diffusion of TLR2 and CD14 on macrophage plasma membrane? 2) Does culturing macrophages on different surface topographies impact the diffusion of TLR2 in the plasma membrane and its pro-inflammatory response, along with morphological changes? 3) Does aging alter the lateral diffusion of TLR2 in the plasma membrane of macrophages? To date, a variety of fluorescence-based methods have been developed to study the dynamics of cell membrane constituents. These techniques are based on either ensemble or single particle measurements. We have used single particle tracking methods to track the mobility of fluorescently labeled membrane receptors on murine bone marrow-derived macrophages. Total internal reflection fluorescence microscopy (TIRF) was used to visualize and capture the dynamics in live cells. Using a custom routine algorithm we detected, localized, and tracked the particles to calculate their diffusion coefficient, extracted from the mean-squared displacement as the most common measure of diffusion. We also measured the diffusion coefficient using an ensemble-based technique known as Raster Image Correlation Spectroscopy (RICS) with a confocal laser-scanning microscope. The use of confocal eliminates the out-of-focus signal and enables measurements that are confined to a narrow plane in the cell. Also, the ability of RICS to separate the slow and immobile fractions of particles makes it possible to detect heterogeneities in diffusion. To our knowledge, this is the first study that has utilized both SPT and RICS to directly compare receptors’ diffusion in different membrane sections. Moreover, this is the first study that has examined the diffusion of receptors on macrophages adhered to different surface topographies, and the first that has investigated the receptors’ diffusion in young and old macrophages. / Thesis / Doctor of Philosophy (PhD) / The immune system is highly dependent on a specialized subset of white blood cells known as macrophages that are capable of clearing damaged and dead cells as well as a wide range of invading micro-organisms. Specific receptor proteins present on the membrane of macrophages are involved in the recognition of particles and subsequent signaling to recruit other immune cells or to promote healing and wound repair. To date, a variety of fluorescence-based microscopy methods have been used to study the dynamics of cell membrane components. The mobility of several membrane receptors in macrophages has been studied using microscopy techniques, which have provided valuable insights into their function. However, there is still insufficient information about the behavior of two key receptors (TLR2 and CD14) that participate in signaling in response to bacterial products. This thesis aims to answer three major questions with regard to receptor mobility (i.e., diffusion) within macrophage membrane: 1) Which type of fluorescence-based microscopy technique is more suitable for measuring the mobility of TLR2 and CD14 receptors on macrophage membranes? 2) What is the impact of different surface topographies on TLR2 diffusion in adhered macrophages, as well as cell shape, and the ability of macrophages to internalize particles? 3) Does aging alter TLR2 mobility in the membrane of macrophages? The following chapters provide detailed answers to these questions. In brief, we have demonstrated that TLR2 and CD14 diffusion measurements in adhered macrophages highly depend on the membrane section chosen. In addition, our results show that micro- and nanostructured surface topographies alter the shape of adhered macrophages and yield higher bacteria internalization, while the diffusion of TLR2 is not changed. When comparing macrophages derived from young and old mice, we find similar diffusion rate of TLR2 in macrophages of the two age groups.

[ES] La calidad superficial en las piezas mecanizadas depende del acabado superficial, resultado de las marcas dejadas por la herramienta durante el proceso de corte. Las aproximaciones teóricas tradicionales indican que estas marcas están relacionadas con los parámetros de corte (velocidad de corte, avance, profundidad de corte...), el tipo de máquina, el material de la pieza, la geometría de la herramienta, etc. Pero no todos los tipos de mecanizado y selección de materiales pueden dar un resultado ambiguo. Hoy en día, de manera progresiva, se están utilizando las técnicas de fresado de Alta Velocidad sobre materiales de difícil mecanizado cada vez más. El fresado de Alta Velocidad implica a un considerable número de parámetros del proceso que pueden afectar a la formación topográfica 3D de la superficie. La hipótesis de que los parámetros de rugosidad superficial dependen de las huellas dejadas por la herramienta, determinadas por las condiciones de trabajo y las propiedades del entorno, condujo al desarrollo de una metodología de investigación personalizada. Este trabajo de investigación muestra como la combinación de los parámetros, inclinación del eje de la herramienta, deflexión geométrica de la herramienta y comportamiento vibracional del entorno, influencian sobre el parámetro de rugosidad superficial 3D, Sz. El modelo general fue dividido en varias partes, donde se ha descrito la influencia de parámetros del proceso adicionales, siendo incluidos en el modelo general propuesto. El proceso incremental seguido permite al autor desarrollar un modelo matemático general, paso a paso, testeando y añadiendo los componentes que más afectan a la formación de la topografía de la superficie. En la primera parte de la investigación se seleccionó un proceso de fresado con herramientas de punta plana. Primero, se analiza la geometría de la herramienta, combinada con múltiples avances, para distinguir los principales parámetros que afectan a la rugosidad superficial. Se introduce un modelo de predicción con un componente básico para la altura de la rugosidad, obtenida por la geometría de la herramienta de corte. A continuación, se llevan a cabo experimentos más específicamente diseñados, variando parámetros tecnológicos. Esto empieza con el análisis de la inclinación del eje de la herramienta contra la mesa de fresado. Los especímenes de análisis son muestras con cuatro recorridos de corte rectos con corte en sentido contrario. Las trayectorias lineales con diferentes direcciones dan la oportunidad de analizar la inclinación del husillo de fresado en la máquina. Un análisis visual reveló diferencias entre direcciones de corte opuestas, así como marcas dejadas por el filo posterior de la herramienta. Considerando las desviaciones de las marcas de corte observadas en las imágenes de rugosidad superficial obtenidas a partir de las medidas, se introdujo un análisis sobre el comportamiento dinámico del equipo y de la herramienta de corte. Las vibraciones producen desviaciones en la mesa de fresado y en la herramienta de corte. Estas desviaciones fueron detectadas e incluidas en el modelo matemático para completar la precisión en la predicción del modelo. Finalmente, el modelo de predicción del parámetro de rugosidad Sz fue comprobado con un mayor número de parámetros del proceso. Los valores de Sz medidos y predichos, fueron comparados y analizados estadísticamente. Los resultados revelaron una mayor desviación de la rugosidad predicha en las muestras fabricadas con diferentes máquinas y con diferentes avances. Importantes conclusiones sobre la precisión del equipo de fabricación han sido extraídas y de ellas se desprende que la huella de la herramienta de corte está directamente relacionada con los parámetros de la topografía de la superficie. Además, la influencia de la huella está afectada por la geometría de la herramienta de corte, la rigidez de la herramienta y la precisión del equipo. La geometría de la herramienta conforma la base del parámetro Sz, desviación de la altura de la superficie. Las conclusiones alcanzadas son la base para recomendaciones prácticas, aplicables en la industria. / [CA] La qualitat superficial en les peces mecanitzades depèn de l'acabat superficial, resultat de les marques deixades per l'eina durant el procés de tall. Les aproximacions teòriques tradicionals indiquen que aquestes marques estan relacionades amb els paràmetres de tall (velocitat de tall, avanç, profunditat de tall...), el tipus de màquina, el material de la peça, la geometria de l'eina, etc. Però no tots els tipus de mecanitzat i selecció de materials poden donar un resultat ambigu. Avui en dia, de manera progressiva, s'estan utilitzant les tècniques de fresat d'Alta Velocitat sobre materials de difícil mecanització cada vegada més. El fresat d'Alta Velocitat implica un considerable nombre de paràmetres del procés que poden afectar la formació topogràfica 3D de la superfície. La hipòtesi que els paràmetres de rugositat superficial depenen de les empremtes deixades per l'eina, determinades per les condicions de treball i les propietats de l'entorn, va conduir al desenvolupament d'una metodologia d'investigació personalitzada. Aquest treball de recerca mostra com la combinació dels paràmetres, inclinació de l'eix de l'eina, deflexió geomètrica de l'eina i comportament vibracional de l'entorn, influencien sobre el paràmetre de rugositat superficial 3D, Sz. El model general va ser dividit en diverses parts, on s'ha descrit la influència de paràmetres addicionals del procés, sent inclosos en el model general proposat. El procés incremental seguit permet a l'autor desenvolupar un model matemàtic general, pas a pas, testejant i afegint els components que més afecten a la formació de la topografia de la superfície. En la primera part de la investigació es va seleccionar un procés de fresat amb eines de punta plana. Primer, s'analitza la geometria de l'eina, combinada amb múltiples avanços, per distingir els principals paràmetres que afecten la rugositat superficial. S'introdueix un model de predicció amb un component bàsic per a l'altura de la rugositat, obtinguda a través de la geometria de l'eina de tall. A continuació, es duen a terme experiments més específicament dissenyats, variant paràmetres tecnològics. Això comença amb l'anàlisi de la inclinació de l'eix de l'eina contra la taula de fresat. Els espècimens d'anàlisi són mostres amb quatre recorreguts de tall rectes amb tall en sentit contrari. Les trajectòries lineals amb diferents direccions donen l'oportunitat d'analitzar la inclinació del fus de fresat en la màquina. Una anàlisi visual revelà diferències entre direccions de tall oposades, així com marques deixades pel tall posterior de l'eina. Considerant les desviacions de les marques de tall observades en les imatges de rugositat superficial obtingudes a partir de les mesures, es va introduir una anàlisi sobre el comportament dinàmic de l'equip i de l'eina de tall. Les vibracions produeixen desviacions en la taula de fresat i en l'eina de tall. Aquestes desviacions van ser detectades i incloses en el model matemàtic per completar la precisió en la predicció de el model. Finalment, el model de predicció de el paràmetre de rugositat Sz va ser comprovat amb un major nombre de paràmetres del procés. Els valors de Sz mesurats i predits, van ser comparats i analitzats estadísticament. Els resultats van revelar una major desviació de la rugositat predita en les mostres fabricades amb diferents màquines i amb diferents avanços. Importants conclusions sobre la precisió de l'equip de fabricació han estat extretes i d'elles es desprèn que l'empremta de l'eina de tall està directament relacionada amb els paràmetres de la topografia de la superfície. A més, la influència de la empremta està afectada per la geometria de l'eina de tall, la rigidesa de l'eina i la precisió de l'equip. La geometria de l'eina conforma la base del paràmetre Sz, desviació de l'altura de la superfície. Les conclusions assolides són la base per recomanacions pràctiques, aplicables en la indústria. / [EN] Surface quality of machined parts highly depends on the surface texture that reflects the marks, left by the tool during the cutting process. The traditional theoretical approaches indicate that these marks are related to the cutting parameters (cutting speed, feed, depths of cut...), the machining type, the part material, the tool geometry, etc. But, different machining type and material selection can give a variable result. In nowadays, more progressively, High Speed milling techniques have been applied on hard-to-cut materials more and more extensively. High-speed milling involves a considerable number of process parameters that may affect the 3D surface topography formation. The hypothesis that surface topography parameters depends on the traces left by the tool, determined by working conditions and environmental properties, led to the development of a custom research methodology. This research work shows how the parameters combination, tool axis inclination, tool geometric deflection, cutting tool geometry and environment vibrational behavior, influence on 3D surface topography parameter Sz. The general model was divided in multiple parts, where additional process parameters influence has been described and included in general model proposed. The incremental process followed allows the author to develop a general mathematical model, step by step, testing and adding the components that affect surface topography formation the most. In the first part of the research a milling procedure with flat end milling tools was selected. First, tool geometry, combined with multiple cutting feed rates, is analyzed to distinguish the main parameters that affect surface topography. A prediction model is introduced with a basic topography height component, performed by cutting tool geometry. Next, specifically designed experiments were conducted, varying technological parameters. That starts with cutting tool axis inclination against the milling table analysis. The specimens of analysis are samples with 4 contrary aimed straight cutting paths. Linear paths in different directions give a chance to analyze milling machine spindle axis topography, as well as marks left from cutting tool back cutting edge. Considering the deviations of cutting marks observed in the images of the surface topography obtained through the measurements, the milling equipment and cutting tool dynamical behavior analysis were introduced. Vibrations produce deviations in the milling table and cutting tool. These deviations were detected and included in the mathematical model to complete the prediction model accuracy. Finally, the prediction model of the topography parameter SZ was tested with increased number of process parameters. Measured and predicted SZ values were compared and analyzed statistically. Results revealed high predicted topography deviation on samples manufactured with different machines and with different feed rates. Relevant conclusions about the manufacturing equipment accuracy have been drawn and they state that cutting tool's footprint is directly related with surface topography parameters. Besides, footprint influence is affected by cutting tool geometry, tool stiffness and equipment accuracy. / Logins, A. (2021). High speed milling technological regimes, process condition and technological equipment condition influence on surface quality parameters of difficult to cut materials [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/164122 / TESIS

Coherence scanning interferometry (CSI) is a non-contacting optical technique which is widely used for the measurement of surface topography. CSI combines the lateral resolution of a high power microscope with the axial resolution of an interferometer. As with any other metrology instrument, CSI is calibrated to define measurement uncertainty. The traditional calibration procedure, as recommended by instrument manufacturers, consists of calibration of the axial and lateral scales of the instrument. Although calibration in this way provides uncertainties for the measurement of rectilinear artefacts, it does not give information about tilt-related uncertainty. If an object with varying slope is measured, significant errors are observed as the surface gradient increases. In this thesis a novel approach of calibration and adjustment for CSI using a spherical object is introduced. This new technique is based on three dimensional linear filtering theory. According to linear theory, smooth surface measurement in CSI can be represented as a linear filtering operation, where the filter is characterised either by point spread function (PSF) in space domain or by transfer function (TF) in spatial frequency domain. The derivation of these characteristics usually involves making the Born approximation, which is strictly only applicable for weakly scattering objects. However, for the case of surface scattering and making use of the Kirchhoff approximation, the system can be considered linear if multiple scattering is assumed to be negligible. In this case, the object is replaced by an infinitely thin foil-like object, which follows the surface topography and, therefore, is called the foil model of the surface. For an ideal aberration free instrument, the linear characteristics are determined by the numerical aperture of the objective lens and the bandwidth of the source. However, it is found that the PSF and TF of a commercial instrument can depart significantly from theory and result in a significant measurement error. A new method, based on modified inverse filter to compensate the phase and amplitude-related errors in the system PSF/TF, is demonstrated. Finally, a method based on de-warping to compensate distortion is discussed. The application of the linear theory as well as modified inverse filter is dependent on the assumption of the shift invariance. As distortion introduces a field dependent magnification, the presence of distortion for CSI with relatively large field of view, restricts the applicability of the linear theory. Along with this restriction, distortion also introduces erroneous height measurement for objects with gradients. This new approach, based on de-warping, resolves the problems associated with distortion.

Traditionally, surface topography measurement was in the domain of quality control of engineering parts. With the advancement of manufacturing technology and affordable computational costs, different types of surfaces are produced with varied shapes and surface textures. These pose significant measurement problems, therefore, surface topography research is gaining momentum to achieve a better control of the surface. Coherence scanning interferometry (CSI) is one of the most common techniques used for measurement of surface topography. It is preferred over tactile and other non-contact techniques since it provides fast and accurate measurement with high vertical (~ 1 nm) and lateral (~1 μm) resolutions over larger areas without any damage to the surface. Essentially, CSI is treated as one dimensional (1D) superposition of the light waves from an object and a reference that generates a three dimensional (3D) interferogram. Secondly, despite the advantages, there is no standard configuration of CSI that can provide absolute surface topography measurement of an engineering part with multiple materials. An effective solution to this problem will be particularly useful in the field of semiconductor and bio-related industries where chips and instruments are made of many materials. In this Thesis, first, the CSI technique is analysed in terms of a wider theoretical framework of 3D linear filtering technique which shows the similarities among other seemingly disparate techniques such as confocal and optical coherence tomography. Due consideration to the spectral characteristic of the source and the effect of numerical aperture are given and important parameters such as vertical and lateral resolutions are computed to compare this theory with standard analysis methods. Additionally, it is shown that the 3D fringe pattern can be considered to be a superposition of a reference field and the scattered field from the top foil-like layer on the top the object. The scattered field from this foil object is dependent on the normal Fresnel reflection coefficients. Therefore, it explains the phase offset and the proportional height offset introduced by different materials, especially, metals. In an object, where multiple materials are present, each material introduces different phase to the fringe pattern and therefore, the surface topography of the entire object is altered. To overcome this problem, the optical polarising properties of the material are exploited. A novel configuration of polarisation sensitive CSI is presented where interferograms with orthogonal circular polarisations are recorded and analysed. The configuration, initially, needs to be calibrated with a material and after that at each point on the object, the refractive index and height offset can be calculated. Therefore, it can be dually used to identify unknown materials present on the object and also to compensate for the height offset introduced by each material to produce absolute surface topography of the entire object. The configuration provides good agreement with ellipsometric results for metals. Additionally, it retains the advantages of high vertical and lateral resolution same as other standard coherence scanning interferometers.

This thesis is focused on the analysis and on the design of the milling technology of the components for the aerospace industry which are characterised by a high aspect ratio, reduced stiffness and all covered by a high demand on the fatigue performance. This work includes the experimental verification of the proposed technology using CNC machining center, evaluation of surface structure using optical high-resolution 3D surface measurement system ALICONA, analysis of the force loading using KISTLER device, fatigue testing and fatigue fracture analysis.

<p>This thesis describes a method of growing a highly adherent nano-porous alumina coating on titanium implant materials, a design which might be useful in hard tissue replacement. Alumina layers were formed by anodisation of aluminium, which had been deposited on titanium and titanium alloys by electron beam evaporation. Mechanical testing showed the coatings’ shear and tensile strength to be ~20MPa and ~10MPa respectively. </p><p>Human osteoblasts were cultured on purchased membranes, produced in the same way with similar characteristics as the coating mentioned above. Cell viability, proliferation and phenotype were assessed by measuring redox reactions, DNA, tritiated thymidine incorporation and alkaline phosphatase production. Results showed normal osteoblastic growth patterns with increasing cell numbers the first two weeks after which cell growth decreased and alkaline phosphatase production increased, indicating that osteoblastic phenotype was retained on the alumina. Flattened cell morphology with filipodia attached to the pores of the material was seen. </p><p>Implants frequently trigger inflammatory responses due to accumulation and activation of cells such as polymorphonuclear granulocytes (PMN), also called neutrophils. Activation and morphology of human PMN in response to nano-porous alumina with two pore sizes (20 and 200 nm) was investigated by luminol-amplified chemiluminescence, granule enzyme deposition measurement, optical and scanning electron microscopy. Activation was observed on both membrane types, however less pronounced on the 200 nm alumina. For both membranes a decrease in activation was seen after coating with fibrinogen, collagen I and serum (more pronounced for the two latter). On fibrinogen-coated alumina many flattened cells were observed, indicating frustrated phagocytosis. Finally when culturing osteoblasts on non-coated and collagen-coated membranes (after exposure to PMN) many more cells had established on the protein-coated surface after 24 h. </p><p>The overall results indicate that it might be possible to produce a novel bone implant coating by anodisation of aluminium deposited on titanium and that this material will support osteoblast adhesion and proliferation. Furthermore neutrophil activation can be suppressed when coating the alumina with collagen I, which is beneficial considering the fact that this protein also is essential for bone formation.</p>

This thesis describes a method of growing a highly adherent nano-porous alumina coating on titanium implant materials, a design which might be useful in hard tissue replacement. Alumina layers were formed by anodisation of aluminium, which had been deposited on titanium and titanium alloys by electron beam evaporation. Mechanical testing showed the coatings’ shear and tensile strength to be ~20MPa and ~10MPa respectively. Human osteoblasts were cultured on purchased membranes, produced in the same way with similar characteristics as the coating mentioned above. Cell viability, proliferation and phenotype were assessed by measuring redox reactions, DNA, tritiated thymidine incorporation and alkaline phosphatase production. Results showed normal osteoblastic growth patterns with increasing cell numbers the first two weeks after which cell growth decreased and alkaline phosphatase production increased, indicating that osteoblastic phenotype was retained on the alumina. Flattened cell morphology with filipodia attached to the pores of the material was seen. Implants frequently trigger inflammatory responses due to accumulation and activation of cells such as polymorphonuclear granulocytes (PMN), also called neutrophils. Activation and morphology of human PMN in response to nano-porous alumina with two pore sizes (20 and 200 nm) was investigated by luminol-amplified chemiluminescence, granule enzyme deposition measurement, optical and scanning electron microscopy. Activation was observed on both membrane types, however less pronounced on the 200 nm alumina. For both membranes a decrease in activation was seen after coating with fibrinogen, collagen I and serum (more pronounced for the two latter). On fibrinogen-coated alumina many flattened cells were observed, indicating frustrated phagocytosis. Finally when culturing osteoblasts on non-coated and collagen-coated membranes (after exposure to PMN) many more cells had established on the protein-coated surface after 24 h. The overall results indicate that it might be possible to produce a novel bone implant coating by anodisation of aluminium deposited on titanium and that this material will support osteoblast adhesion and proliferation. Furthermore neutrophil activation can be suppressed when coating the alumina with collagen I, which is beneficial considering the fact that this protein also is essential for bone formation.

The trend for designing of a titanium implant explored using different chemical compositions and crystallinity materials until people realized that the implant surface character was another important factor affecting the rate and extent of osseointegartion. Titanium received a macroporous titania surface layer by anodization, which contains open pores with average pore diameter around 5μm. An additional mesoporous titania top layer was created that followed the contour of the macropores and having 100–200 nm thickness and a pore diameter of 10 nm. Thus, a coherent laminar titania surface layer was obtained producing a hierarchical macro- and mesoporous surface. The interfacial bonding between the surface layers and the titanium matrix was characterized by a scratch test that confirmed a stable and strong bonding of the laminar titania surface layers upon titanium. The wettability to water and the effects on the osteosarcoma cell line (SaOS-2) proliferation and mineralization of the formed titania surface layers were studied systematically by cell culture and scanning electron microscopy (SEM). A synergistic role of the hierarchical macro- and mesoporosities was revealed in terms of enhancing cell adhesion, proliferation and mineralization, when compared with the titania surface with solo porosity scale topography. For the in vivo results of the evaluation of osseointegration, an argon ion beam polishing technique was applied to prepare the cross sections of implants feasible for the high resolution SEM investigation. The interfacial microstructure between newly formed bone and implants with four modified surfaces including the new hierarchical macro- and mesoporous implant surface retrieved after in vivo tests were characterized. By this approach it has become possible to directly observe early bone formation, the increase of bone density, and the evolution of bone structure. The two bone growth mechanisms, distant osteogenesis and contact osteogenesis, can also be distinguished. These direct observations give, at microscopic level, a better view of osseointegration and explain the functional mechanisms of various implant surfaces for osseointegration. / <p>At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 3: In press. Paper 4: Manuscript.</p>

No description available.

Basal cell carcinoma is a form of a non-melanoma skin tumour, that commonly forms over the sun exposed regions of the head and neck. Investigation of the rate of occurrence at different sites on face and neck shows considerable variation from site to site. The inner canthus has a disproportionate number when compared to more exposed sites. The eye brow ridge, cheek bone and nose limit the field of view of the inner canthus, thus it is expected to receive less radiation than other more exposed regions. To explain the disproportionate rate, it is hypothesised that a portion of radiation incident onto the eye is reflected to the inner canthus. The aim of this thesis is to investigate the contribution that the radiation reflected off the surface of the eye makes to the overall dose on the inner canthus. The inter reflections between the eye and inner canthus were studied through the use of the ray tracing program Zemax. Zemax was used to trace rays in a non sequential mode incident onto a model eye and periorbital region. To obtain the models of the eye and periorbital region, both magnetic resonance imaging and a casting process was investigated, with the later being superior for our uses. With the model obtained, it was used in a series of three dimensional ray tracing programs. On a macroscopic scale there is a small increase in the irradiance on the inner canthus (2 % over a 1 cm2 area). Peaks of high irradiance (19 % increase in irradiance above direct irradiance) were discovered over the surface when the detector was divided into 200 mm elements. It was concluded that these increases above the direct irradiance in these small regions, increases the possibility of the occurrence of a Basal cell carcinoma. Individual facial geometry, will greatly effect the location and size of these peaks and as a result an experimental method to measure the dose distribution across the inner canthus was proposed. Initially it was planned to use polysulphone film to measure the erythemal dose on the inner canthus. Results from the modelling indicated that any measurements made had to be at a high spatial resolution. Polysulphone film was found to be inadequate for this, due to its large uncertainties. An alternative method was investigated so that a population study could be performed in future studies using visible radiation and high dynamic range images gave a simple and effective clinical assessment tool. The high dynamic range images showed hot spots in the irradiance across the inner canthus agreeing with the model. The small spots of high relative irradiance may not be the only reason for the increased rate in this region. Greater skin sensitivity and absence of sun screen use at this site are other possibilities. It is believed however that the irradiance distribution across the inner canthus on a microscopic scale goes a long way to increasing the risk for certain people.