Development and regeneration of neuronal circuits in the vertebrate retina

Thesis (Ph.D.)--University of Washington, 2016-08

Logic-Based Delivery of Site-Specifically-Modified Proteins from Gels through Engineered Biomacromolecular Architecture

Thesis (Master's)--University of Washington, 2016-08

Design and Fabrication of Perovskite Micro-Cavity Lasers

Thesis (Master's)--University of Washington, 2016-08

Involving individuals in the manufacturing strategy formation : Strategic consensus among workers and managers

Decisions made and actions taken by individuals in the operations function impact the formation of a company’s manufacturing strategy (MS). Therefore, it is important that the MS is understood and agreed on by all employees, that is, strategic consensus among the individuals in the operations function is essential. This research contributes to the current body of knowledge by including a workers’ perspective on MS formation. It is the workers on the shop floor who bring the MS to life in the actual operations through their daily decisions and actions. The MS falls short if the priorities outlined do not materialise in practice as intended. The purpose of this research is to investigate how the individuals in the operations function perceive the MS in order to understand how these individuals are involved in the MS formation. The research is based on five studies, differing by evidence, as follows: one theoretical, three qualitative in the setting of small and medium-sized enterprises (SMEs), and one quantitative at a large company. Based on the findings presented in the six appended papers, the results show that empirically and conceptually, workers have been overlooked or given a passive role in the MS formation. Empirically, it is seen that workers and managers do not have a shared understanding of the underlying reasons for strategic priorities; hence, the level of strategic consensus is low. Furthermore, the level of strategic consensus varies among the different MS dimensions depending on their organisational level. Moreover, the empirical findings reveal that internal contextual factors influence the individuals’ perceptions of the MS and the possibilities for strategic consensus. Regarding the external context, the results show that major customers’ strategies influence the subcontractor SMEs’ MS formation. The usage of means of communication in the operations function has also shown to be of importance for how the MS is perceived. Conceptually, the findings indicate that the MS literature tends to treat individuals in the operations function in a deterministic manner; individuals on the shop floor are regarded as manufacturing resources. To ensure a successful MS formation process, where the patterns of the decisions made by the individuals in the operations function forms the MS, the view on human nature within the MS requires a more voluntaristic approach. This research suggests to view the MS formation as an iterative “patterning process” which builds on a reciprocal relationship between workers and managers. The introduction of the patterning process contributes to the research on MS formation by explaining the perception range within the hierarchical levels, by re-defining the hierarchical levels included in the MS formation and by detailing the activities in the MS formation.

dachshund Potentiates Hedgehog Signaling during Drosophila Retinogenesis

Proper organ patterning depends on a tight coordination between cell proliferation and differentiation. The patterning of Drosophila retina occurs both very fast and with high precision. This process is driven by the dynamic changes in signaling activity of the conserved Hedgehog (Hh) pathway, which coordinates cell fate determination, cell cycle and tissue morphogenesis. Here we show that during Drosophila retinogenesis, the retinal determination gene dachshund (dac) is not only a target of the Hh signaling pathway, but is also a modulator of its activity. Using developmental genetics techniques, we demonstrate that dac enhances Hh signaling by promoting the accumulation of the Gli transcription factor Cubitus interruptus (Ci) parallel to or downstream of fused. In the absence of dac, all Hh-mediated events associated to the morphogenetic furrow are delayed. One of the consequences is that, posterior to the furrow, dac- cells cannot activate a Roadkill-Cullin3 negative feedback loop that attenuates Hh signaling and which is necessary for retinal cells to continue normal differentiation. Therefore, dac is part of an essential positive feedback loop in the Hh pathway, guaranteeing the speed and the accuracy of Drosophila retinogenesis.

Identification and functional analysis of novel genes expressed in the Anterior Visceral Endoderm

During early vertebrate development, the correct establishment of the body axes is critical. The anterior pole of the mouse embryo is established when Distal Visceral Endoderm (DVE) cells migrate to form the Anterior Visceral Endoderm (AVE). Symmetrical expression of Lefty1, Cer1 and Dkk1 determines the direction of DVE migration and the future anterior side. In addition to the establishment of the Anterior-Posterior axis, the AVE has also been implicated in anterior neural specification. To better understand the role of the AVE in these processes, we have performed a differential screening using Affymetrix GeneChip technology with AVE cells isolated from cer1P-EGFP transgenic mouse embryos. We found 175 genes which were upregulated in the AVE and 36 genes in the Proximal-posterior sample. Using DAVID software, we characterized the AVE cell population regarding cellular component, molecular function and biological processes. Among the genes that were found to be upregulated in the AVE, several novel genes were identified. Four of these transcripts displaying high-fold change in the AVE were further characterized by in situ hybridization in early stages of development in order to validate the screening. From those four selected genes, one, denominated Adtk1, was chosen to be functionally characterized by targeted inactivation in ES cells. Adtk1 encodes for a serine/threonine kinase. Adtk1 null mutants are smaller and present short limbs due to decreased mineralization, suggesting a potential role in chondrogenesis during limb development. Taken together, these data point to the importance of reporting novel genes present in the AVE.

Bioanalytical applications of microfluidic devices

The first part of the thesis describes a new patterning technique--microfluidic contact printing--that combines several of the desirable aspects of microcontact printing and microfluidic patterning and addresses some of their important limitations through the integration of a track-etched polycarbonate (PCTE) membrane. Using this technique, biomolecules (e.g., peptides, polysaccharides, and proteins) were printed in high fidelity on a receptor modified polyacrylamide hydrogel substrate. The patterns obtained can be controlled through modifications of channel design and secondary programming via selective membrane wetting. The protocols support the printing of multiple reagents without registration steps and fast recycle times. The second part describes a non-enzymatic, isothermal method to discriminate single nucleotide polymorphisms (SNPs). SNP discrimination using alkaline dehybridization has long been neglected because the pH range in which thermodynamic discrimination can be done is quite narrow. We found, however, that SNPs can be discriminated by the kinetic differences exhibited in the dehybridization of PM and MM DNA duplexes in an alkaline solution using fluorescence microscopy. We combined this method with multifunctional encoded hydrogel particle array (fabricated by stop-flow lithography) to achieve fast kinetics and high versatility. This approach may serve as an effective alternative to temperature-based method for analyzing unamplified genomic DNA in point-of-care diagnostic.

Dissecting signaling pathways that control glia migration in the developing Drosophila retina

The function of a complex nervous system relies on an intricate interaction between neurons and glial cells. However, as glial cells are generally born distant from the place where they settle, molecular cues are important to direct their migration. Glial cell migration is important in both normal development and disease, thus current research in the laboratory has been focused on dissecting regulatory events underlying that crucial process. With this purpose, the Drosophila eye imaginal disc has been used as a model. In response to neuronal photoreceptor differentiation, glial cells migrate from the CNS into the eye disc where they act to correctly wrap axons. To ensure proper development, attractive and repulsive signals must coordinate glial cell migration. Importantly, one of these signals is Bnl, a Fibroblast Growth Factor (FGF) ligand expressed by retinal progenitor cells that was suggested to act as a non-autonomous negative regulator of excessive glial cell migration (overmigration) by binding and activating the Btl receptor expressed by glial cells. Through the experimental results described in chapter 3 we gained a detailed insight into the function of bnl in eye disc growth, photoreceptor development, and glia migration. Interestingly, we did not find a direct correlation between the defects on the ongoing photoreceptors and the glia overmigration phenotype; however, bnl knockdown caused apoptosis of eye progenitor cells what was strongly correlated with glia migration defects. Glia overmigration due to Bnl down-regulation in eye progenitor cells was rescued by inhibiting the pro-apoptotic genes or caspases activity, as well as, by depleting JNK or Dp53 function in retinal progenitor cells. Thus, we suggest a cross-talk between those developmental signals in the control of glia migration at a distance. Importantly, these results suggest that Bnl does not control glial migration in the eye disc exclusively through its ability to bind and activate its receptor Btl in glial cells. We also discuss possible biological roles for the glia overmigration in the bnl knockdown background. Previous results in the lab showed an interaction between dMyc, a master regulator of tissue growth, and Dpp, a Transforming Growth Factor-β important for retinal patterning and for accurate glia migration into the eye disc. Thus, we became interested in understanding putative relationships between Bnl and dMyc. In chapter 4, we show that they positively cooperate in order to ensure proper development of the eye disc. This work highlights the importance of the FGF signaling in eye disc development and reveals a signaling network where a range of extra- and intra-cellular signals cooperate to non-autonomously control glial cell migration. Therefore, such inter-relations could be important in other Drosophila cellular contexts, as well as in vertebrate tissue development.

MOLECULAR MASS MANIPULATION ENABLED BY GRAPHENE BASED NANOSTRUCTURES

The surge of interest in graphene, as epitomized by the Nobel Prize in Physics in 2010, is attributed to its extraordinary properties. Graphene is ultrathin, mechanically tough, and has amendable surface chemistry. These features make graphene and graphene based nanostructure an ideal candidate for the use of molecular mass manipulation. The controllable and programmable molecular mass manipulation is crucial in enabling future graphene based applications, however is challenging to achieve. This dissertation studies several aspects in molecular mass manipulation including mass transportation, patterning and storage. For molecular mass transportation, two methods based on carbon nanoscroll are demonstrated to be effective. They are torsional buckling instability assisted transportation and surface energy induced radial shrinkage. To achieve a more controllable transportation, a fundamental law of direction transport of molecular mass by straining basal graphene is studied. For molecular mass patterning, we reveal a barrier effect of line defects in graphene, which can enable molecular confining and patterning in a domain of desirable geometry. Such a strategy makes controllable patterning feasible for various types of molecules. For molecular mass storage, we propose a novel partially hydrogenated bilayer graphene structure which has large capacity for mass uptake. Also the mass release can be achieved by simply stretching the structure. Therefore the mass uptake and release is reversible. This kind of structure is crucial in enabling hydrogen fuel based technology. Lastly, spontaneous nanofluidic channel formation enabled by patterned hydrogenation is studied. This novel strategy enables programmable channel formation with pre-defined complex geometry.

Cell-cell communication in three dimensional microenvironments

Cellular behavior is dependent on a variety of extracellular cues required for normal tissue function, wound healing, and activation of the immune system. Removed from their in vivo microenvironment and cultured in vitro, cells lose many environmental cues and that may result in abberant behavior, making it difficult to study cellular processes. In order to mimic native tissue environments, optical tweezer and microfluidic technologies were used to place cells within defined areas of the culture environment. To provide three dimensional supports found in natural tissues, hydrogel scaffolds of poly (ethylene glycol) diacrylate and the basement membrane matrix Matrigel were used. Optical tweezer technology allowed precision placement and formation of homotypic and heterotypic arrays of human U937, HEK 293, and porcine mesenchymal stem cells. Alternatively, two microfluidic devices were designed to pattern Matrigel scaffolds. The first microfluidic device utilized laminar flow to spatially pattern multiple cell types within the device. Gradients of soluble molecules were then be formed and manipulated across the Matrigel scaffolds. Patterning Matrigel using laminar flow techniques require microfluidic expertise and do not produce consistent patterning conditions, limiting their use difficult in most cell culture laboratories. Thus, a buried Matrigel polydimethylsiloxane (PDMS) device was developed for spatial patterning of biological scaffolds. Matrigel is injected into micron sized channels of PDMS fabricated by soft lithography and allowed to thermally cure. Following curing, a second PDMS device was placed on top of the buried Matrigel channels to support media flow. In order to validate these systems, a cell-cell communication model system was developed utilizing LPS and TNFα signaling with fluorescent reporter systems to monitor communication in real time. We demonstrated the utility of microfluidic devices to support the cell-cell communication model system by co culturing three cell types within Matrigel scaffolds and monitoring signaling activity via fluorescent reporters.

Activation of Sonic hedgehog signaling in ventricular cardiomyocytes exerts cardioprotection against ischemia reperfusion injuries

International audience

Sonic hedgehog signaling regulates mode of cell division of early cerebral cortex progenitors and increases

The morphogen Sonic Hedgehog (SHH) plays a critical role in the development of different tissues. In the central nervous system, SHH is well known to contribute to the patterning of the spinal cord and separation of the brain hemispheres. In addition, it has recently been shown that SHH signaling also contributes to the patterning of the telencephalon and establishment of adult neurogenic niches. In this work, we investigated whether SHH signaling influences the behavior of neural progenitors isolated from the dorsal telencephalon, which generate excitatory neurons and macroglial cells in vitro. We observed that SHH increases proliferation of cortical progenitors and generation of astrocytes, whereas blocking SHH signaling with cyclopamine has opposite effects. In both cases, generation of neurons did not seem to be affected. However, cell survival was broadly affected by blockade of SHH signaling. SHH effects were related to three different cell phenomena: mode of cell division, cell cycle length and cell growth. Together, our data in vitro demonstrate that SHH signaling controls cell behaviors that are important for proliferation of cerebral cortex progenitors, as well as differentiation and survival of neurons and astroglial cells.

Is phraseology the third articulation of language? Fresh insights into a theoretical conundrum

Is phraseology the third articulation of language? Fresh insights into a theoretical conundrum Jean-Pierre Colson University of Louvain (Louvain-la-Neuve, Belgium) Although the notion of phraseology is now used across a wide range of linguistic disciplines, its definition and the classification of phraseological units remain a subject of intense debate. It is generally agreed that phraseology implies polylexicality, but this term is problematic as well, because it brings us back to one of the most controversial topics in modern linguistics: the definition of a word. On the other hand, another widely accepted principle of language is the double articulation or duality of patterning (Martinet 1960): the first articulation consists of morphemes and the second of phonemes. The very definition of morphemes, however, also poses several problems, and the situation becomes even more confused if we wish to take phraseology into account. In this contribution, I will take the view that a corpus-based and computational approach to phraseology may shed some new light on this theoretical conundrum. A better understanding of the basic units of meaning is necessary for more efficient language learning and translation, especially in the case of machine translation. Previous research (Colson 2011, 2012, 2013, 2014), Corpas Pastor (2000, 2007, 2008, 2013, 2015), Corpas Pastor & Leiva Rojo (2011), Leiva Rojo (2013), has shown the paramount importance of phraseology for translation. A tentative step towards a coherent explanation of the role of phraseology in language has been proposed by Mejri (2006): it is postulated that a third articulation of language intervenes at the level of words, including simple morphemes, sequences of free and bound morphemes, but also phraseological units. I will present results from experiments with statistical associations of morphemes across several languages, and point out that (mainly) isolating languages such as Chinese are interesting for a better understanding of the interplay between morphemes and phraseological units. Named entities, in particular, are an extreme example of intertwining cultural, statistical and linguistic elements. Other examples show that the many borrowings and influences that characterize European languages tend to give a somewhat blurred vision of the interplay between morphology and phraseology. From a statistical point of view, the cpr-score (Colson 2016) provides a methodology for adapting the automatic extraction of phraseological units to the morphological structure of each language. The results obtained can therefore be used for testing hypotheses about the interaction between morphology, phraseology and culture. Experiments with the cpr-score on the extraction of Chinese phraseological units show that results depend on how the basic units of meaning are defined: a morpheme-based approach yields good results, which corroborates the claim by Beck and Mel'čuk (2011) that the association of morphemes into words may be similar to the association of words into phraseological units. A cross-linguistic experiment carried out for English, French, Spanish and Chinese also reveals that the results are quite compatible with Mejri’s hypothesis (2006) of a third articulation of language. Such findings, if confirmed, also corroborate the notion of statistical semantics in language. To illustrate this point, I will present the PhraseoRobot (Colson 2016), a computational tool for extracting phraseological associations around key words from the media, such as Brexit. The results confirm a previous study on the term globalization (Colson 2016): a significant part of sociolinguistic associations prevailing in the media is related to phraseology in the broad sense, and can therefore be partly extracted by means of statistical scores. References Beck, D. & I. Mel'čuk (2011). Morphological phrasemes and Totonacan verbal morphology. Linguistics 49/1: 175-228. Colson, J.-P. (2011). La traduction spécialisée basée sur les corpus : une expérience dans le domaine informatique. In : Sfar, I. & S. Mejri, La traduction de textes spécialisés : retour sur des lieux communs. Synergies Tunisie n° 2. Gerflint, Agence universitaire de la Francophonie, p. 115-123. Colson, J.-P. (2012). Traduire le figement en langue de spécialité : une expérience de phraséologie informatique. In : Mogorrón Huerta, P. & S. Mejri (dirs.), Lenguas de especialidad, traducción, fijación / Langues spécialisées, figement et traduction. Encuentros Mediterráneos / Rencontres Méditerranéennes, N°4. Universidad de Alicante, p. 159-171. Colson, J.-P. (2013). Pratique traduisante et idiomaticité : l’importance des structures semi-figées. In : Mogorrón Huerta, P., Gallego Hernández, D., Masseau, P. & Tolosa Igualada, M. (eds.), Fraseología, Opacidad y Traduccíon. Studien zur romanischen Sprachwissenschaft und interkulturellen Kommunikation (Herausgegeben von Gerd Wotjak). Frankfurt am Main, Peter Lang, p. 207-218. Colson, J.-P. (2014). La phraséologie et les corpus dans les recherches traductologiques. Communication lors du colloque international Europhras 2014, Association Européenne de Phraséologie. Université de Paris Sorbonne, 10-12 septembre 2014. Colson, J-P. (2016). Set phrases around globalization : an experiment in corpus-based computational phraseology. In: F. Alonso Almeida, I. Ortega Barrera, E. Quintana Toledo and M. Sánchez Cuervo (eds.), Input a Word, Analyse the World: Selected Approaches to Corpus Linguistics. Newcastle upon Tyne: Cambridge Scholars Publishing, p. 141-152. Corpas Pastor, G. (2000). Acerca de la (in)traducibilidad de la fraseología. In: G. Corpas Pastor (ed.), Las lenguas de Europa: Estudios de fraseología, fraseografía y traducción. Granada: Comares, p. 483-522. Corpas Pastor, G. (2007). Europäismen - von Natur aus phraseologische Äquivalente? Von blauem Blut und sangre azul. In: M. Emsel y J. Cuartero Otal (eds.), Brücken: Übersetzen und interkulturelle Kommunikationen. Festschrift für Gerd Wotjak zum 65. Geburtstag, Fráncfort: Peter Lang, p. 65-77. Corpas Pastor, G. (2008). Investigar con corpus en traducción: los retos de un nuevo paradigma [Studien zur romanische Sprachwissenschaft und interkulturellen Kommunikation, 49], Fráncfort: Peter Lang. Corpas Pastor, G. (2013). Detección, descripción y contraste de las unidades fraseológicas mediante tecnologías lingüísticas. In Olza, I. & R. Elvira Manero (eds.) Fraseopragmática. Berlin: Frank & Timme, p. 335-373. Leiva Rojo, J. (2013). La traducción de unidades fraseológicas (alemán-español/español-alemán) como parámetro para la evaluación y revisión de traducciones. In: Mellado Blanco, C., Buján, P, Iglesias N.M., Losada M.C. & A. Mansilla (eds), La fraseología del alemán y el español: lexicografía y traducción. ELS, Etudes Linguistiques / Linguistische Studien, Band 11. München: Peniope, p. 31-42. Leiva Rojo, J. & G. Corpas Pastor (2011). Placing Italian idioms in a foreign milieu: a case study. In: Pamies Bertrán, A., Luque Nadal, L., Bretana, J. &; M. Pazos (eds), (2011). Multilingual phraseography. Second Language Learning and Translation Applications. Baltmannsweiler: Schneider Verlag (Colección: Phraseologie und Parömiologie, 28), p. 289-298. Martinet, A. (1966). Eléments de linguistique générale. Paris: Colin. Mejri, S. (2006). Polylexicalité, monolexicalité et double articulation. Cahiers de Lexicologie 2: 209-221.

The development of planar high-K/III-V p-channel MOSFETs for post-silicon CMOS

Conventional Si complementary-metal-oxide-semiconductor (CMOS) scaling is fast approaching its limits. The extension of the logic device roadmap for future enhancements in transistor performance requires non-Si materials and new device architectures. III-V materials, due to their superior electron transport properties, are well poised to replace Si as the channel material beyond the 10nm technology node to mitigate the performance loss of Si transistors from further reductions in supply voltage to minimise power dissipation in logic circuits. However several key challenges, including a high quality dielectric/III-V gate stack, a low-resistance source/drain (S/D) technology, heterointegration onto a Si platform and a viable III-V p-metal-oxide-semiconductor field-effect-transistor (MOSFET), need to be addressed before III-Vs can be employed in CMOS. This Thesis specifically addressed the development and demonstration of planar III-V p-MOSFETs, to complement the n-MOSFET, thereby enabling an all III-V CMOS technology to be realised. This work explored the application of InGaAs and InGaSb material systems as the channel, in conjunction with Al2O3/metal gate stacks, for p-MOSFET development based on the buried-channel flatband device architecture. The body of work undertaken comprised material development, process module development and integration into a robust fabrication flow for the demonstration of p-channel devices. The parameter space in the design of the device layer structure, based around the III-V channel/barrier material options of In_x≥0.53Ga_1-xAs/In_0.52Al_0.48As and In_x≥0.1Ga_1-xSb/AlSb, was systematically examined to improve hole channel transport. A mobility of 433 cm²/Vs, the highest room temperature hole mobility of any InGaAs quantum-well channel reported to date, was obtained for the In_0.85Ga_0.15As (2.1% strain) structure. S/D ohmic contacts were developed based on thermally annealed Au/Zn/Au metallisation and validated using transmission line model test structures. The effects of metallisation thickness, diffusion barriers and de-oxidation conditions were examined. Contacts to InGaSb-channel structures were found to be sensitive to de-oxidation conditions. A fabrication process, based on a lithographically-aligned double ohmic patterning approach, was realised for deep submicron gate-to-source/drain gap (L_side) scaling to minimise the access resistance, thereby mitigating the effects of parasitic S/D series resistance on transistor performance. The developed process yielded gaps as small as 20nm. For high-k integration on GaSb, ex-situ ammonium sulphide ((NH₄)₂S) treatments, in the range 1%-22%, for 10min at 295K were systematically explored for improving the electrical properties of the Al₂O₃/GaSb interface. Electrical and physical characterisation indicated the 1% treatment to be most effective with interface trap densities in the range of 4 - 10×10¹²cm^-2eV^-1 in the lower half of the bandgap. An extended study, comprising additional immersion times at each sulphide concentration, was further undertaken to determine the surface roughness and the etching nature of the treatments on GaSb. A number of p-MOSFETs based on III-V-channels with the most promising hole transport and integration of the developed process modules were successfully demonstrated in this work. Although the non-inverted InGaAs-channel devices showed good current modulation and switch-off characteristics, several aspects of performance were non-ideal; depletion-mode operation, modest drive current (I_d,sat=1.14mA/mm), double peaked transconductance (g_m=1.06mS/mm), high subthreshold swing (SS=301mV/dec) and high on-resistance (R_on=845kΩ.μm). Despite demonstrating substantial improvement in the on-state metrics of I_d,sat (11×), g_m (5.5×) and R_on (5.6×), inverted devices did not switch-off. Scaling gate-to-source/drain gap (L_side) from 1μm down to 70nm improved I_d,sat (72.4mA/mm) by a factor of 3.6 and gm (25.8mS/mm) by a factor of 4.1 in inverted InGaAs-channel devices. Well-controlled current modulation and good saturation behaviour was observed for InGaSb-channel devices. In the on-state In_0.3Ga_0.7Sb-channel (I_d,sat=49.4mA/mm, g_m=12.3mS/mm, R_on=31.7kΩ.μm) and In_0.4Ga_0.6Sb-channel (I_d,sat=38mA/mm, g_m=11.9mS/mm, R_on=73.5kΩ.μm) devices outperformed the InGaAs-channel devices. However the devices could not be switched off. These findings indicate that III-V p-MOSFETs based on InGaSb as opposed to InGaAs channels are more suited as the p-channel option for post-Si CMOS.

Africans in Scotland: heterogeneity and sensitivities to HIV

Aim: To investigate how diversity within the African migrant population in Scotland affects their understandings of HIV and uptake of HIV testing and treatment, in order to improve HIV-related outcomes. Background: In the UK, Africans have the worst outcomes for HIV infection, primarily due to late diagnosis. Improvement requires better understanding of the barriers to healthcare engagement. This PhD study investigates how diversity among first generation African migrants in Scotland could affect engagement with general healthcare and HIV related interventions and services. Methods: I conducted qualitative research, involving participant observation at two sites (an African religious group and an asylum seeker/refugee drop-in centre) and interviews with African migrants attending these and three additional sites (two advocacy charities and a student association). Data were collected in two cities (Glasgow and Edinburgh) and two smaller towns (Paisley and Kirkcaldy). I interviewed 27 Africans, including economic migrants (n=8), students (n=9) and asylum seeker/refugees (n=10) and 14 representatives from organisations with high levels of African attendees (e.g., country associations, community organisations, advocacy groups, commercial establishments and religious based organisations). Thematic data analysis was carried out. Results: Diversity of the population and related issues of identity: Participants were highly diverse and reported considerable heterogeneity in the African diaspora in Scotland. The identity of “African” was bound with various negative stereotypes and appeals to this identity did not necessarily have relevance for participants. Nature of African affiliated organisations in Scotland: There were a wide range of organisations that advertised their remit as catering for the African diaspora. They varied in consistency and sustainability and contributed towards healthcare engagement to different degrees. Engagement with healthcare: There were multiple experiences and understandings of the healthcare system within the sample as a whole, and to an extent by migrant type. Whilst the majority reported successful and satisfactory service use, distinct barriers emerged. These included: understandings of rights and access to care based on African models of healthcare; the interplay of religious based understandings with ideas about access to healthcare; and assumptions and anxiety about the connections between visa status and health status. Knowledge of HIV and engagement with HIV related services: Participants had good knowledge about HIV, with some notable exceptions, but there was no patterning by migrant type. They had diverse views about risk of HIV infection, most of which did not align with the HIV epidemiology that identifies African migrants as an at risk group. Most of the sample did not think targeting African migrants for HIV interventions would be successful and were hostile to the proposal for various reasons, especially because they believed it would perpetuate stigma and prejudice towards the African diaspora. There were mixed experiences of HIV related services, and prompts to test for HIV had elicited a range of reactions, the majority negative. Conclusion: Diversity within the African diaspora in Scotland should be taken into account to improve the salience and relevance of future HIV interventions. Attitudes towards current HIV testing promotion suggest that a more cooperative approach could be taken with African communities to build on existing relationships of trust and understandings of HIV.