Use of confocal and multiphoton microscopy for the evaluation of micro-optical components and emitters

We report on the application of confocal and multiphoton microscopic techniques for the evaluation of the latest generation of micro optical components. The optical emitting characteristics of arrays of matrix addressable GaN micrometer-sized light emitting diodes (micro-LEDs) have been measured using a commercial confocal microscope utilising the LEDs' own emission along with reflection confocal microscopy to determine the surface structure. Multiphoton induced luminescence from the 10-20-micron diameter emitters has also been used to examine the structure of the device and we compare this with electrically induced emission. In related work, the optical properties of micro lens arrays (10-100-micron diameter) fabricated in SiC, Sapphire, and Diamond have been determined using transmission confocal microscopy. Such optical microscopy techniques offer a simple, non-destructive method to determine the structure and performance of such novel devices

Nano handling and measurement of biological cells in culture

A thesis submitted to the University of Bedfordshire in partial fulfillment of the requirements for the degree of Doctor of PhilosophyThis thesis systematically investigates the nano handling and measurement techniques for biological cells in culture and studies the techniques to realize innovative and multi-functional applications in biomedicine. Among them, the technique based on AFM is able to visualize and quantify the dynamics of organic cells in culture on the nano scale. Especially, the cellular shear adhesion force on the various locations of biological cells was firstly accurately measured in the research of the cell-substrate interaction in terms of biophysical perspective. The innovative findings are conductive to study the cell-cell adhesion, the cell-matrix adhesion which is related to the cell morphology structure, function, deformation ability and adhesion of cells and better understand the cellular dynamic behaviors. Herein, a new liquid-AFM probe unit and an increment PID control algorithm were implemented suitable for scanning the cell samples under the air conditions and the liquid environments. The influence between the surface of sample and the probe, and the damage of probe during the sample scanning were reduced. The proposed system is useful for the nano handling and measurement of living cells. Besides, Besides, to overcome the limitations of liquid-AFMs, the multiple optical tweezers were developed to integrate with the liquid-AFM. The technique based on laser interference is able to characterize the optical trap stiffness and the escape velocity, especially to realize the capture and sorting of multiple cells by a polarization-controlled periodic laser interference. It can trap and move hundreds of cells without physical contact, and has broad application prospects in cytology. Herein, a new experimental method integrated with the positioning analysis in the Z direction was used to improve the fluid force method for the calibration and characterize the mechanical forces exerted on optical traps and living cells. Moreover, a sensitive and highly efficient polarization-controlled three-beam interference set-up was developed for the capture and sorting of multiple cells. By controlling the polarization angles of the beams, various intensity distributions and different sizes of dots were obtained. Subsequently, we have experimentally observed multiple optical tweezers and the sorting of cells with different polarization angles, which are in accordance with the theoretical analysis

Magnetometry with nitrogen-vacancy defects in diamond

The isolated electronic spin system of the Nitrogen-Vacancy (NV) centre in diamond offers unique possibilities to be employed as a nanoscale sensor for detection and imaging of weak magnetic fields. Magnetic imaging with nanometric resolution and field detection capabilities in the nanotesla range are enabled by the atomic-size and exceptionally long spin-coherence times of this naturally occurring defect. The exciting perspectives that ensue from these characteristics have triggered vivid experimental activities in the emerging field of "NV magnetometry". It is the purpose of this article to review the recent progress in high-sensitivity nanoscale NV magnetometry, generate an overview of the most pertinent results of the last years and highlight perspectives for future developments. We will present the physical principles that allow for magnetic field detection with NV centres and discuss first applications of NV magnetometers that have been demonstrated in the context of nano magnetism, mesoscopic physics and the life sciences.Comment: Review article, 28 pages, 16 figure

Current methods for characterising mixing and flow in microchannels

This article reviews existing methods for the characterisation of mixing and flow in microchannels, micromixers and microreactors. In particular, it analyses the current experimental techniques and methods available for characterising mixing and the associated phenomena in single and multiphase flow. The review shows that the majority of the experimental techniques used for characterising mixing and two-phase flow in microchannels employ optical methods, which require optical access to the flow, or off-line measurements. Indeed visual measurements are very important for the fundamental understanding of the physics of these flows and the rapid advances in optical measurement techniques, like confocal scanning laser microscopy and high resolution stereo micro particle image velocimetry, are now making full field data retrieval possible. However, integration of microchannel devices in industrial processes will require on-line measurements for process control that do not necessarily rely on optical techniques. Developments are being made in the areas of non-intrusive sensors, magnetic resonance techniques, ultrasonic spectroscopy and on-line flow through measurement cells. The advances made in these areas will certainly be of increasing interest in the future as microchannels are more frequently employed in continuous flow equipment for industrial applications

DNA builds and strengthens the extracellular matrix in Myxococcus xanthus biofilms by interacting with exopolysaccharides.

One intriguing discovery in modern microbiology is the extensive presence of extracellular DNA (eDNA) within biofilms of various bacterial species. Although several biological functions have been suggested for eDNA, including involvement in biofilm formation, the detailed mechanism of eDNA integration into biofilm architecture is still poorly understood. In the biofilms formed by Myxococcus xanthus, a Gram-negative soil bacterium with complex morphogenesis and social behaviors, DNA was found within both extracted and native extracellular matrices (ECM). Further examination revealed that these eDNA molecules formed well organized structures that were similar in appearance to the organization of exopolysaccharides (EPS) in ECM. Biochemical and image analyses confirmed that eDNA bound to and colocalized with EPS within the ECM of starvation biofilms and fruiting bodies. In addition, ECM containing eDNA exhibited greater physical strength and biological stress resistance compared to DNase I treated ECM. Taken together, these findings demonstrate that DNA interacts with EPS and strengthens biofilm structures in M. xanthus

Development of a controlled environment near-field optical microscope for organic thin film studies

The requirements of a microscope differ greatly from field to field and with the sample being studied Organic thin films are highly sensitive to environmental conditions being rapidly photobleached in the presence of oxygen In this paper we introduce the design of a low cost near field optical microscope suitable for environmentally controlled studies. We demonstrate that reducing ambient pressure to similar to 10 mTorr (1 3 Pa) of nitrogen (a pressure accessible with a mechanical pump) significantly reduces photo-bleaching on a typical experimental timescale. In addition, we introduce a novel non-optical feedback control mechanism combining a piezoelectric diaphragm with a tuning fork. The morphology of thin films of MEH-PPV spin cast from the polymer dissolved in chlorobenzene is investigated These films are shown to be featureless on a 100 nm length scale in terms of topology, PL intensity and PL spectrum both before and during photo-bleaching. As far as we are aware these are the first reported thin films of MEH-PPV grown in which similar to 100 nm sized domains arc absent. We believe that the ability to control environment will not only allow better optical characterization of thin films but also opens up the possibility of the production of novel two dimensional organic photonic crystals through a combination of mechanical disruption, photo-bleaching and photo-conjugation of polymers

Cavity-enhanced Raman Microscopy of Individual Carbon Nanotubes

Raman spectroscopy reveals chemically specific information and provides label-free insight into the molecular world. However, the signals are intrinsically weak and call for enhancement techniques. Here, we demonstrate Purcell enhancement of Raman scattering in a tunable high-finesse microcavity, and utilize it for molecular diagnostics by combined Raman and absorption imaging. Studying individual single-wall carbon nanotubes, we identify crucial structural parameters such as nanotube radius, electronic structure and extinction cross-section. We observe a 320-times enhanced Raman scattering spectral density and an effective Purcell factor of 6.2, together with a collection efficiency of 60%. Potential for significantly higher enhancement, quantitative signals, inherent spectral filtering and absence of intrinsic background in cavity-vacuum stimulated Raman scattering render the technique a promising tool for molecular imaging. Furthermore, cavity-enhanced Raman transitions involving localized excitons could potentially be used for gaining quantum control over nanomechanical motion and open a route for molecular cavity optomechanics

Micro- and sub-microstructuring and characterisation of technical surfaces by means of laser direct writing including a novel approach for laser beam profiling

Within recent years, numerous fields of engineering, like mechanics, optics and electronics, have been influenced and revolutionised by the technique of microand nano-structuring. For example, special optical elements for beam shaping, surface structures for the reduction of friction or modern "lab on chip" devices have been produced. Within this thesis a universal system has been developed facilitating the production of such structured surfaces with dimensions down to 500 nm. This system is not only capable of structuring surfaces by means of lithographic processes; it further allows the inspection of surfaces by scanning their topography. To realise such a system, two different technologies have been evaluated: Scanning Near-field Optical Lithography (SNOL), a very sophisticated technique which uses a thin fibre tip to expose a photo resist-covered surface, and confocal scanning technology. Here, the confocal scanning is accomplished using an adapted optical component, the optical pickup unit (OPU), from a gaming console, which turned out to be the most suitable and cost-efficient solution for the realisation of this system. Several test series have been carried out during this work, to verify the performance of the confocal system, both to structure photo resist surfaces and to characterise unknown surfaces. This present work will show the ability of the developed system to produce structures down to the sub-micron range and to characterise unknown surfaces with sub- micron precision. Various patterns have been written into photo resistcoated substrates to structure their surface. Beginning with diffractive optical elements (DOE) for beam shaping, followed by Dammann gratings for twodimensional beam shaping and optical gratings for light guidance as well as producing technical surfaces imitating the properties of sharkskin or simple micromechanical structures, the developed confocal system has shown itself to be flexible and widely-applicable. IV During the development of the confocal system, a strong need for a beam profiling system analysing the light beam diverging from the OPU, was recognised. Due to the fact that no commercially available system was capable of characterising beam sizes within the range of the diffraction limit, a novel method for beam profiling was invented. This method makes use of the fibre tips already applied within the SNOL system, producing tomographical scans of the beam spot