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Enhancing productivity in bioprocesses, especially for biofuel production, is crucial for achieving an environmentally and economically sustainable biotechnology industry.Metabolic channelling occurs in nature when the intermediate between two consecutive enzymes in a pathway is directed from the first enzyme to the second avoiding diffusion in the cytosol. This would be very advantageous in bioprocesses as it would increase efficiency of a particular pathway, reducing side products and protecting the cells from potential toxic intermediates. In recent years different strategies for emulating channelling effect wereproposed and used with very promising results. Clustering of enzymes seems to be the simplest way to create metabolic channelling. In this master thesis, four different strategies to co-localize enzymes in clusters are compared. The metabolic pathway chosen as a model was ethanol production by pyruvate decarboxylase (Pdc) and alcohol dehydrogenase (Adh). Chimeric proteins were genetically engineered and transformed in E. coli creating different strains. Ethanol production by the different strains was measured to compare production efficiency. Cell growth and protein expression were used for further understanding of the results. Strengths and weaknesses of each strategy and proposals for further improvement were discussed.

Les modes de Laguerre-Gauss sont une des solutions de l’équation de propagation de la lumière dans l’approximation paraxiale en coordonnées cylindriques. Ces modes sont caractérisés par deux indices (azimutal et radial) et présentent des propriétés particulières : une structure en forme d’anneau et une phase en hélice. Ces propriétés ont été mises au profit pour plusieurs applications allant de la microscopie à l’astronomie. Cette thèse a été dédiée à l’étude de ces modes et à leur application dans le domaine des atomes froids. Dans un premier temps, la pureté des modes fabriqués par la méthode d’holographie numérique a été étudiée, ainsi que le rôle que la pureté joue dans leur propagation. Ces modes ont été ensuite utilisés pour l’obtention une source d’atomes froids brillante. Depuis plus de 20 ans, des efforts considérables ont été faits pour obtenir des sources d’atomes froids les plus brillantes et les plus compactes possibles. Un piège magnéto-optique à deux dimensions (2D-MOT) est couramment utilisé pour obtenir une source continue d’atomes froids avec un flux important de l’ordre de 1010 atomes/s. Toutefois, le jet atomique ainsi obtenu présente une divergence d’environ 40 mrad ce qui contraint l’utilisateur à travailler près de la sortie mais avec un accès optique limité ou bien loin où il a un bon accès optique mais au prix d’une densité atomique plus faible. Une alternative est présentée, dans laquelle un mode de Laguerre-Gauss est utilisé pour canaliser les atomes à la sortie d’un 2D-MOT. Le mode de Laguerre-Gauss réalise un piégeage dipolaire qui confine les atomes sont le centre noir du mode, ce qui présente l’intérêt de limiter le chauffage dû à l’absorption/émission de la lumière. Ainsi, puisque les modes de Laguerre-Gauss gardent leur forme au cours de leur propagation, les atomes sont canalisés sur une distance de plusieurs décimètres. Nous avons étudié le fonctionnement de ce système pour différents ordres du mode de Laguerre-Gauss et différentes fréquences, et nous avons montré que ce système est efficace et permet d’atteindre un gain en densité d’un facteur 200 par rapport au cas d’un 2D-MOT conventionnel. Un cas particulier est aussi présenté, dans lequel la fréquence du mode de Laguerre-Gauss est choisie pour effectuer non seulement la canalisation des atomes mais aussi leur repompage, ce qui conduit à une simplification importante du système. D’un autre côté, les modes de Laguerre-Gauss sont d’un grand intérêt dans le domaine de l’information et la cryptographie quantique car ils peuvent être utilisés pour encoder et enregistrer l’information. Pour cela, il est indispensable de pouvoir les détecter de façon non-équivoque. Dans ce contexte, nous nous sommes intéressés à la détermination de ces modes. Jusqu’à maintenant, les techniques de détection ont permis de mesurer l’indice azimutal mais peu d’entre elles mesurent l’indice radial. Si on est capable de mesurer aussi l’indice radial, il peut être utilisé comme une nouvelle variable pour le codage et l’enregistrement de l’information. Sous cette motivation, une technique basée sur la transformation du mode de Laguerre-Gauss par un système astigmatique a été mise au point pour déterminer les deux indices qui le caractérisent. Nous avons montré que cette technique peut aussi être utilisée pour optimiser expérimentalement la fabrication des modes de Laguerre-Gauss d’ordre élevé les plus purs possibles. / Laguerre-Gaussian modes of light are one of the solutions to the propagation equation of light in the paraxial approximation in cylindrical coordinates. These modes are characterized by two indices (azimuthal and radial) and present particular properties: ring-shaped structure and a helical phase. These properties have been put to use in several applications going from microscopy to astronomy. This work has been devoted to the study of these modes and their application in the cold atom domain. Initially, the purity of such modes generated by numerical holography was studied, as well as the role played by the purity in their propagation. These modes were then used to obtain a bright source of cold atoms. For more than 20 years, considerable efforts have been made to create sources of cold atoms as bright and compact as possible. A two dimensional magneto-optical trap (2D-MOT) is currently used to obtain a continuous source of cold atoms with high flux of the order of 1010 atomes/s. Nevertheless, the source of cold atoms thereby achieved show a divergence of about 40 mrad which constrains the user to work by output but with a limited optical access or far from the output but at the cost of lower density. An alternative is presented, in which a Laguerre-Gaussian mode of light is used to channel the atoms at the output of a 2D-MOT. The Laguerre-Gaussian mode of light carries out a dipole trapping that confines the atoms in the black center of the mode, which has the advantage of limiting the heating due to absorption/emission of light. Thus, since the Laguerre-Gaussian modes of light keep their ring-shape along their propagation, the atoms are channeled over a distance of several decimeters. We studied the functioning of this system for different orders of the Laguerre-Gaussian mode and at different frequencies, and we showed that this system is efficient and allows to achieve a density gain of a factor 200 compared to a conventional 2D-MOT. A particular case is also presented, in which the frequency of the Laguerre-Gaussian mode of light is chosen to carry out not only the channeling of the atoms, but also their repumping, which leads to an important simplification of the system. Laguerre-Gaussian modes of light are also of great interest in the domain of quantum information and quantum cryptography because they can be used to encode and store information. In order to do that, it is crucial to be able to detect them in an unequivocal way. In this context, we are interested in the determination of these modes. Until now, different detection techniques have measured the azimuthal index, but few of them have been able to measure the radial index. If we are able to measure the radial index, it can be used as a new variable to encode and store information. Under this motivation, a technique based on the transformation of a Laguerre-Gaussian mode of light by an astigmatic system has been extended to determine the two indices characterizing them. We showed that this technique can also be used to experimentally optimize the generation of the purest possible Laguerre-Gaussian modes of high order.

Cette thèse se décompose en deux parties : i) l’étude du creusement d'un canal dans un plasma sous-dense (0.1nc<n<nc, nc étant la densité critique) de plusieurs centaines de microns par une impulsion laser de durée 1-10 ps et d'intensité 10^18 à 10^20 W/cm² ; ii) les mécanismes de saturation de la diffusion Raman arrière stimulée d'une impulsion laser de durée ps et d'intensité 10^14 à 10^16 W/cm². Le creusement d’un canal plasma par un laser très intense a fait l’objet d’une étude paramétrique à l’aide d’un code PIC (Particle In Cell) 2D. On obtient différents types de canaux en fonction des paramètres du laser et du plasma, reproduisant ainsi et élargissant des résultats précédent. De plus, la vitesse de creusement du canal a été mesurée, et des lois d’échelle ont été établies pour les plasmas homogènes. Elles sont ensuite appliquées à des plasmas inhomogènes, du type de ceux rencontrés lors de la fusion par confinement inertiel (FCI). Cela permet de prévoir l’énergie nécessaire pour creuser un canal jusqu’à la densité critique, étape importante de la FCI par allumage rapide. La saturation du Raman a été étudiée d'un point de vue numérique, pour déterminer si la cause de la saturation était due au déphasage ou à la croissance d'une onde satellite (« sideband »), en utilisant diverses approches. La première est de regarder les résultats de simulations Raman (donc électromagnétiques) à partir de codes cinétiques PIC et Vlasov. La deuxième, consiste à regarder ce qui se passe lorsque l'on initialise un plasma avec une fonction de distribution issue de la théorie adiabatique à l'aide d'un code Vlasov (donc dans une version purement électrostatique). Dans ce cas, on observe bien la croissance d'une onde satellite, dont le nombre d'onde dominant, ainsi que le taux de croissance sont en bon accord avec ce que l'on observe dans les simulations cinétiques. Au final, la saturation de l’onde plasma peut être causée par les deux mécanismes de saturation. / This thesis is divided in two parts : i) the laser channeling in hundreds of microns long under-dense plasmas (0.1nc<n<nc, nc being the critical density) of a laser pulse of intensity 10^18 to 10^20 W/cm² and duration 1-10 ps; ii) the saturation mechanisms of stimulated Raman back-scattering of a laser pulse of intensity 10^14 to 10^16 W/cm² and duration of about 1 ps. A parametric study was performed to study the channeling of a very intense laser pulse, using a 2D PIC (Particle In Cell) code. Various kinds of channels were obtained depending on the laser and plasma parameters, thereby reproducing and enlarging previous studies. Moreover, the channeling velocity was measured and scaling laws were established for homogeneous plasmas. They are then applied to inhomogeneous plasmas, similar to those encountered in inertial confinement fusion (ICF). It is then possible to estimate the energy necessary to channel to the critical density, an important step for the fast ignition scheme of ICF. Raman saturation was studied using numerical simulations, in order to determine if it is due to dephasing or to the growth of sidebands, using different approaches. The first is to study Raman simulations (electromagnetic) performed with kinetic PIC and Vlasov codes. The second, is to study the evolution of a plasma initialized with a distribution function after the adiabatic theory, using a Vlasov code (electrostatic). In this case, we observe the growth of a sideband, with dominant wavenumber and growth rate in good agreement with kinetic simulations. The saturation of the plasma wave can be caused by both saturation mechanisms.

<p>Three different studies have been made; an analysis of the general weather situation at three weather stations in Svalbard, close to the main settlement Longyearbyen; an instrumental comparison between three weather stations operating at nearly the same place during a few days in October 2006; and a case study of a weather station at the mountain Breinosa compared to the station in the valley beneath.</p><p>The analysis of the general weather at the three stations, Adventdalen (data used from 2004-2006), Svalbard Airport (data used from 1994-2004) and Gruvefjellet (data used from 2002-2005) showed that it is clear that the local surroundings do contribute to a more locally produced weather. The mean wind speed in Adventdalen was shown to be 5.1 m/s and the wind direction was mostly from inland towards the coast. Channelling effects dominates the winds in the valley, resulting in high wind speeds. The channelled winds from Adventdalen often reach Svalbard Airport (average wind speed 4.8 m/s). At the Airport there is also a contribution of winds (mostly in summer) originating from the sea, e.g. sea breeze. In times when the weather at Svalbard Airport was more continental, Adventfjorden and Isfjorden were likely to be covered by ice. The higher location of Gruvefjellet most often contributes to low temperatures and an average wind speed of 3.9 m/s. However, the temperature at Adventdalen and the Airport are often colder than at Gruvefjellet due to the frequency of temperature inversions.</p><p>The instrumental comparison concerned three stations in Adventdalen. Data from the stationary weather station was compared with data from two temporarily stations during 4-7th October 2006. It is apparent that the stationary station needs calibration, at least when looking at the pressure measurements, where there clearly is an offset in the data.</p><p>The third study was a case study concerned data from a station at the mountain Breinosa and Adventdalen. A new weather station will be put up during 2007 and data (16th December 2005 until 12th January 2006) from a temporarily station was used when determining the differences from the station down in the valley (Adventdalen). The wind speed is often just as high in the valley as at Breinosa due to channelling effects. The temperature in Adventdalen seems to be affected by the temperature at Breinosa. The air is chilled and sinks down to be channelled through the valley. There are some occasions when the temperature at the mountain is higher than beneath. During those events the wind speed is low and a temperature inversion develops at the ground.</p> / <p>Tre olika studier genomförts; en analysering av data från tre väderstationer på Svalbard;en instrumentell jämförelse mellan tre väderstationer på Svalbard nära huvudorten Longyearbyen; en fallstudie av data från en väderstation på berget Breinosa jämfört med situationen i Adventdalen nedanför.</p><p>Undersökningen av generella vädersituationen vid de tre stationerna, Adventdalen (data från 2004-2006), Svalbard Flygplats (data från 1994-2004) och Gruvefjellet (data från 2002-2005) visade tydligt att den lokala omgivningen bidrar mycket till hur vädret formas. Medelvindhastigheten i Adventdalen var 5,1 m/s och vindriktningen mestadels längs dalen ut mot fjorden. Kanaliseringseffekter dominerar vindarna i dalen och bidrar till de höga vindhastigheterna. Kanaliserade vindar från Adventdalen når ofta Svalbard Flygplats (medel vindhastighet 4,8 m/s). Vid Flygplatsen finns också ett visst bidrag från vindar utifrån havet (mestadels under sommaren), d.v.s. sjöbris. I situationer då vädret vid flygplatsen varit mer kontinentalt så har Adventfjorden mest troligt varit täckt av is.</p><p>Gruvefjellets höga placering bidrar till låga temperaturer och en medelvindhastighet på 3.9 m/s. Dock är temperaturen är ofta lägre i Adventdalen och vid Flygplatsen på grund av att det ofta förekommer temperaturinversioner i marknivå.</p><p>Den instrumentella jämförelsen berörde tre stationer i Adventdalen. Data från den stationära stationen jämfördes under 4-7 oktober 2006 med data från två temporära stationer. Det är tydligt att den stationära stationen är i behov av kalibrering. I alla fall är det uppenbart vid studien av lufttrycket som tydligt är förskjutet och visar ett lägre tryck.</p><p>Tredje studien var en fallstudie och data undersöktes från berget Breinosa och Adventdalen. En ny väderstation kommer att monteras på Breinosa under 2007 och data (16 december 2005 till 12 januari 2006) från en temporär station användes i studien. Vindhastigheten är ofta lika hög i dalen som på berget på grund av kanaliseringseffekter. Temperaturen i dalen förefaller vara påverkad av temperatursituationen på berget. Avkyld luft sjunker och kanaliseras ner i dalen. Det finns några tillfällen då temperaturen på berget Breinosa är högre än nere i dalen. Under de situationerna avtar vindhastigheten och en temperaturinversion bildas i marknivå.</p>

Three different studies have been made; an analysis of the general weather situation at three weather stations in Svalbard, close to the main settlement Longyearbyen; an instrumental comparison between three weather stations operating at nearly the same place during a few days in October 2006; and a case study of a weather station at the mountain Breinosa compared to the station in the valley beneath. The analysis of the general weather at the three stations, Adventdalen (data used from 2004-2006), Svalbard Airport (data used from 1994-2004) and Gruvefjellet (data used from 2002-2005) showed that it is clear that the local surroundings do contribute to a more locally produced weather. The mean wind speed in Adventdalen was shown to be 5.1 m/s and the wind direction was mostly from inland towards the coast. Channelling effects dominates the winds in the valley, resulting in high wind speeds. The channelled winds from Adventdalen often reach Svalbard Airport (average wind speed 4.8 m/s). At the Airport there is also a contribution of winds (mostly in summer) originating from the sea, e.g. sea breeze. In times when the weather at Svalbard Airport was more continental, Adventfjorden and Isfjorden were likely to be covered by ice. The higher location of Gruvefjellet most often contributes to low temperatures and an average wind speed of 3.9 m/s. However, the temperature at Adventdalen and the Airport are often colder than at Gruvefjellet due to the frequency of temperature inversions. The instrumental comparison concerned three stations in Adventdalen. Data from the stationary weather station was compared with data from two temporarily stations during 4-7th October 2006. It is apparent that the stationary station needs calibration, at least when looking at the pressure measurements, where there clearly is an offset in the data. The third study was a case study concerned data from a station at the mountain Breinosa and Adventdalen. A new weather station will be put up during 2007 and data (16th December 2005 until 12th January 2006) from a temporarily station was used when determining the differences from the station down in the valley (Adventdalen). The wind speed is often just as high in the valley as at Breinosa due to channelling effects. The temperature in Adventdalen seems to be affected by the temperature at Breinosa. The air is chilled and sinks down to be channelled through the valley. There are some occasions when the temperature at the mountain is higher than beneath. During those events the wind speed is low and a temperature inversion develops at the ground. / Tre olika studier genomförts; en analysering av data från tre väderstationer på Svalbard;en instrumentell jämförelse mellan tre väderstationer på Svalbard nära huvudorten Longyearbyen; en fallstudie av data från en väderstation på berget Breinosa jämfört med situationen i Adventdalen nedanför. Undersökningen av generella vädersituationen vid de tre stationerna, Adventdalen (data från 2004-2006), Svalbard Flygplats (data från 1994-2004) och Gruvefjellet (data från 2002-2005) visade tydligt att den lokala omgivningen bidrar mycket till hur vädret formas. Medelvindhastigheten i Adventdalen var 5,1 m/s och vindriktningen mestadels längs dalen ut mot fjorden. Kanaliseringseffekter dominerar vindarna i dalen och bidrar till de höga vindhastigheterna. Kanaliserade vindar från Adventdalen når ofta Svalbard Flygplats (medel vindhastighet 4,8 m/s). Vid Flygplatsen finns också ett visst bidrag från vindar utifrån havet (mestadels under sommaren), d.v.s. sjöbris. I situationer då vädret vid flygplatsen varit mer kontinentalt så har Adventfjorden mest troligt varit täckt av is. Gruvefjellets höga placering bidrar till låga temperaturer och en medelvindhastighet på 3.9 m/s. Dock är temperaturen är ofta lägre i Adventdalen och vid Flygplatsen på grund av att det ofta förekommer temperaturinversioner i marknivå. Den instrumentella jämförelsen berörde tre stationer i Adventdalen. Data från den stationära stationen jämfördes under 4-7 oktober 2006 med data från två temporära stationer. Det är tydligt att den stationära stationen är i behov av kalibrering. I alla fall är det uppenbart vid studien av lufttrycket som tydligt är förskjutet och visar ett lägre tryck. Tredje studien var en fallstudie och data undersöktes från berget Breinosa och Adventdalen. En ny väderstation kommer att monteras på Breinosa under 2007 och data (16 december 2005 till 12 januari 2006) från en temporär station användes i studien. Vindhastigheten är ofta lika hög i dalen som på berget på grund av kanaliseringseffekter. Temperaturen i dalen förefaller vara påverkad av temperatursituationen på berget. Avkyld luft sjunker och kanaliseras ner i dalen. Det finns några tillfällen då temperaturen på berget Breinosa är högre än nere i dalen. Under de situationerna avtar vindhastigheten och en temperaturinversion bildas i marknivå.

Microbial degradation of aromatic hydrocarbons is imperative for maintaining the global carbon cycle and removing potentially toxic aromatic xenobiotics. This thesis focuses on the characterization of a pyruvate-specific class II aldolase (BphI) and acetaldehyde dehydrogenase (BphJ), the final two enzymes of the bph meta-cleavage pathway in Burkholderia xenovorans LB400. This pathway is responsible for the degradation of the industrial pollutant polychlorinated biphenyls (PCB) and therefore mechanistic characterization of these enzymes can be applied to improve pollutant degradation. BphI catalyzes the aldol cleavage of 4-hydroxy-2-oxoacids to pyruvate and an aldehyde while BphJ transforms aldehydes to acyl-CoA, using NAD+ and CoASH as cofactors. Size-exclusion chromatography was used to determine that the oligomeric unit of the BphI-BphJ complex is a heterotetramer. The aldolase BphI was shown to exhibit a compulsory order mechanism and utilize 4-hydroxy-2-oxoacids with an S configuration at C4. The generation of BphI active site variants allowed for the proposal of a catalytic mechanism and a greater understanding as to how stereospecificity occurs. Using steady-state kinetic assays, Arg-16 was demonstrated to be essential for catalysis. Molecular modeling of the substrate and pH dependency (wild-type pKa of ~7, lost in H20A and H20S variants) were used to identify His-20 as the catalytic base. Tyr-290 was originally proposed to be the catalytic acid. However, this was refuted as a Tyr-290 (Y290F) variant did not affect the catalytic efficiency of the enzyme. Instead, the variant was observed to exhibit a loss of stereochemical control. From the crystal structure of an orthologous aldolase-dehydrogenase complex, solvent isotope effect studies, and a proton inventory, a water molecule was implicated as the catalytic acid. Based on their position within the crystal structure, Leu-87 and Leu-89 were implicated in substrate specificity. Replacement of Leu-89 with alanine effectively increased the length of the active site, allowing for the accommodation of longer aldehyde substrates. In contrast, Leu-87 was responsible for hydrophobic stabilization of the C4-methyl of the substrate. Double variants L87N;Y290F and L87W;Y290F were constructed to enable the binding of 4(R)-hydroxy-2-oxoacids. Polarimetric analysis confirmed that the double variants were able to synthesize 4-hydroxy-2-oxoacids of up to 8 carbons in lengths, which were of the opposite stereoisomer to those produced by the wild-type enzyme. Cys-131 was identified as the catalytic thiol that forms an acyl-enzyme intermediate in the dehydrogenase, BphJ. This enzyme was shown to exhibit similar specificity constants for acetaldehyde and propionaldehyde and utilize aliphatic aldehydes from two to five carbons in length as substrates. The enzyme was able to use either NAD+ or NADP+ as the cofactor. Finally, we demonstrated that aldehydes produced in the aldolase reaction are not released into the bulk solvent but are channeled directly to the dehydrogenase, providing the first biochemical determination of substrate channeling in any aldolase-dehydrogenase complex. / Chapter 3 - Reprinted (adapted) with permission from Baker, P., Carere, J., and Seah, S. Y. (2011) Probing the Molecular Basis of Substrate Specificity, Stereospecificity, and Catalysis in the Class II Pyruvate Aldolase, BphI, Biochemistry 50: 3559-3569. Copyright (2011) American Chemical Society. Chapter 4 - Reprinted (adapted) with permission from Baker, P., and Seah, S. Y. (2012) Rational design of stereoselectivity in the class II pyruvate aldolase BphI, J Am Chem Soc 134: 507-513. Copyright (2012) American Chemical Society. Chapter 6 - Reprinted (adapted) with permission from Baker, P., Hillis, C., Carere, J., and Seah, S. Y. (2012) Protein-protein interactions and substrate channeling in orthologous and chimeric aldolase-dehydrogenase complexes, Biochemistry 51: 1942-1952. Copyright (2012) American Chemical Society. / National Science and Engineering Research Council of Canada (NSERC), Ontario Graduate Scholarship in Science and Technology

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Hochaufgelöste räumlich-selektive Einzelionenimplantation ist eine Schlüsseltechnologie um Festkörper-Qubits herzustellen. Der in dieser Arbeit verwendete Nanoimplanter benutzt zur Kollimation eines niederenergetischen Ionenstrahls auf Nanometerebene eine Rasterkraftmikroskop-(AFM-)Spitze, welche mit einer Nanoapertur ausgestattet ist. Diese Technik wurde bereits für verschiedene Quantenanwendungen genutzt. In dieser Arbeit wird sie auf die Erzeugung strukturierter Stickstoff-Fehlstellen-(NV-)Zentren weiterentwickelt und optimiert. Dies umfasst unter anderem die Installation eines neuen AFM-Systems, welches den Aufbau mit zwei nützlichen Funktionen aufrüstet: die In-situ-Aperturvermessung und die Untersuchung von Ionen-sensitiven Fotolacken. Weiter werden die zwei wichtigsten limitierenden Faktoren der räumlichen Auflösung durch Simulationen und Experimente detailliert untersucht. Die Ergebnisse geben Aufschluss über optimale Nanoaperturen und Implantationsbedingungen. Streueffekte an der AFM-Spitze und Gitterführungen in Diamant können dadurch maßgeblich reduziert werden. Weiter werden NV-limitierende Effekte durch mehrere Ausheizschritte sowie Ionen- und Elektronenbestrahlungen untersucht. Zuletzt werden erstmals diamantbasierte Ionendetektoren hergestellt, welche mit Kapazität- und Strom-Spannungs-Messungen, durch Röntgenbestrahlung und Ionenstrahl-induzierter Ladung (IBIC) charakterisiert werden. Die Ergebnisse zeigen, dass die angefertigten Detektoren die Bedingungen für eine deterministische Implantation erfüllen, so dass dieses Prinzip zukünftig in den Nanoimplanter integriert werden kann. / High-resolution spatial-selective single ion implantation is a key technology to produce solid state qubits. The nanoimplanter used in this work collimates a low-energy ion beam at the nanometer level using an atomic force microscope (AFM) tip, which is provided with a nanoaperture. This technique has already been used for various quantum applications. In this thesis it is further developed and optimized for the generation of structured nitrogen vacancy (NV) centers. This includes the installation of a new AFM system, which upgrades the setup with two useful functions: in-situ aperture measurement and the investigation of ion sensitive photoresists. Furthermore, the two most significant limiting factors of spatial resolution are studied in detail by simulations and experiments. The results indicate optimized nanoapertures and implantation conditions. Scattering effects at the AFM tip and ion channeling in diamond can be significantly reduced. Moreover, NV-limiting effects are investigated by several heating steps as well as ion and electron irradiations. Finally, novel diamond based ion detectors are manufactured, that are characterized by capacitance and current-voltage measurements, by X-ray irradiation and ion beam induced charge (IBIC). The results show these detectors fulfill the conditions for a deterministic implantation, so that this concept can be integrated into the nanoimplanter in the future.