Smart imaging using laser targeting: a multiple barcodes application

To the best of our knowledge, proposed is a novel variable depth of field smart imager design using intelligent laser targeting for high productivity multiple barcodes reading applications. System smartness comes via the use of an Electronically Controlled Variable Focal-Length Lens (ECVFL) to provide an agile pixel (and/or pixel set) within the laser transmitter and optical imaging receiver. The ECVFL in the receiver gives a flexible depth of field that allows clear image capture over a range of barcode locations. Imaging of a 660 nm wavelength laser line illuminated 95-bit one dimensional barcode is experimentally demonstrated via the smart imager for barcode target distances ranging from 10 cm to 54 cm. The smart system captured barcode images are evaluated using a proposed barcode reading algorithm. Experimental results after computer-based post-processing show a nine-fold increase in barcode target distance variation range (i.e., range variation increased from 2.5 cm to 24.5 cm) when compared to a conventional fixed lens imager. Applications for the smart imager include industrial multiple product tracking, marking, and inspection system

Tomography applied to Lamb wave contact scanning nondestructive evaluation

The aging world-wide aviation fleet requires methods for accurately predicting the presence of structural flaws that compromise airworthiness in aircraft structures. Nondestructive Evaluation (NDE) provides the means to assess these structures quickly, quantitatively, and noninvasively. Ultrasonic guided waves, Lamb waves, are useful for evaluating the plate and shell structures common in aerospace applications. The amplitude and time-of-flight of Lamb waves depend on the material properties and thickness of a medium, and so they can be used to detect any areas of differing thickness or material properties which indicate flaws. By scanning sending and receiving transducers over an aircraft, large sections can be evaluated after a single pass. However, while this technique enables the detection of areas of structural deterioration, it does not allow for the quantification of the extent of that deterioration. Tomographic reconstruction with Lamb waves allows for the accurate reconstruction of the variation of quantities of interest, such as thickness, throughout the investigated region, and it presents the data as a quantitative map. The location, shape, and extent of any flaw region can then be easily extracted from this Tomographic image. Two Lamb wave tomography techniques using Parallel Projection tomography (PPT) and Cross Borehole tomography (CBT), are shown to accurately reconstruct flaws of interest to the aircraft industry. A comparison of the quality of reconstruction and practicality is then made between these two methods, and their limitations are discussed and shown experimentally. Higher order plate theory is used to derive analytical solutions for the scattering of the lowest order symmetric Lamb wave from a circular inclusion, and these solutions are used to explain the scattering effects seen in the Tomographic reconstructions. Finally, the means by which this scattering theory can be used to develop Lamb wave Tomographic algorithms that are more generally applicable in-the-field, is presented

Development and application of fluorescence lifetime imaging and super-resolution microscopy

This PhD thesis reports the development and application of fluorescence imaging technologies for studying biological processes on spatial scales below the diffraction limit. Two strategies were addressed: firstly fluorescence lifetime imaging (FLIM) to study molecular processes, e.g. using Förster resonance energy transfer (FRET) to read out protein interactions, and secondly direct imaging of nanostructure using super-resolution microscopy (SRM). For quantitative FRET readouts, the development and characterisation of an automated multiwell plate FLIM microscope for high content analysis (HCA) is described. Open source software was developed for the data acquisition and analysis, and approaches to improve the performance of time-gated imaging for FLIM were evaluated including different methods to despeckle the laser illumination and testing of an enhanced detector. This instrument was evaluated using standard fluorescent dye samples and cells expressing fluorescent protein-based FRET constructs. It was applied to an assay of live cells expressing a FRET biosensor and to FRET readouts of aggregation of a membrane receptor (DDR1) in fixed cells. A novel instrument, combining structured illumination microscopy (SIM) with FLIM, was developed to explore the combination of SRM and FLIM-FRET readouts. This enabled the simultaneous mapping of molecular readouts with FLIM and super-resolved imaging. The SIM+FLIM system was applied to image collagen-stimulated DDR1 aggregation in cells, to image DNA structures during the cell cycle and to explore interactions between cell organelles. A novel SRM approach based on a stimulated emission of depletion (STED) microscope incorporating a spatial light modulator (SLM) was developed to provide straightforward robust alignment with collinear excitation/depletion beams, aberration correction, an extended field of view and multiple beam scanning for faster STED image acquisition. The performance of easySLM-STED was evaluated by imaging bead samples, labelled vimentin in Vero cells and the synaptonemal complex in homologs of C. elegans germlines.Open Acces

Deep Tissue Light Delivery and Fluorescence Tomography with Applications in Optogenetic Neurostimulation

Study of the brain microcircuits using optogenetics is an active area of research. This method has a few advantages over the conventional electrical stimulation including the bi-directional control of neural activity, and more importantly, specificity in neuromodulation. The first step in all optogenetic experiments is to express certain light sensitive ion channels/pumps in the target cell population and then confirm the proper expression of these proteins before running any experiment. Fluorescent bio-markers, such as green fluorescent protein (GFP), have been used for this purpose and co-expressed in the same cell population. The fluorescent signal from such proteins provides a monitory mechanism to evaluate the expression of optogenetic opsins over time. The conventional method to confirm the success in gene delivery is to sacrifice the animal, retract and slice the brain tissue, and image the corresponding slices using a fluorescent microscope. Obviously, determining the level of expression over time without sacrificing the animal is highly desirable. Also, optogenetics can be combined with cell-type specific optical recording of neural activity for example by imaging the fluorescent signal of genetically encoded calcium indicators. One challenging step in any optogenetic experiment is delivering adequate amount of light to target areas for proper stimulation of light sensitive proteins. Delivering sufficient light density to a target area while minimizing the off-target stimulation requires a precise estimation of the light distribution in the tissue. Having a good estimation of the tissue optical properties is necessary for predicting the distribution of light in any turbid medium. The first objective of this project was the design and development of a high resolution optoelectronic device to extract optical properties of rats\u27 brain tissue (including the absorption coefficient, scattering coefficient, and anisotropy factor) for three different wavelengths: 405nm, 532nm and 635nm and three different cuts: transverse, sagittal, and coronal. The database of the extracted optical properties was linked to a 3D Monte Carlo simulation software to predict the light distribution for different light source configurations. This database was then used in the next phase of the project and in the development of a fluorescent tomography scanner. Based on the importance of the fluorescent imaging in optogenetics, another objective of this project was to design a fluorescence tomography system to confirm the expression of the light sensitive proteins and optically recording neural activity using calcium indicators none or minimally invasively. The method of fluorescence laminar optical tomography (FLOT) has been used successfully in imaging superficial areas up to 2mm deep inside a scattering medium with the spatial resolution of ~200µm. In this project, we developed a FLOT system which was specifically customized for in-vivo brain imaging experiments. While FLOT offers a relatively simple and non-expensive design for imaging superficial areas in the brain, still it has imaging depth limited to 2mm and its resolution drops as the imaging depth increases. To address this shortcoming, we worked on a complementary system based on the digital optical phase conjugation (DOPC) method which was shown previously that is capable of performing fluorescent tomography up to 4mm deep inside a biological tissue with lateral resolution of ~50 µm. This system also provides a non-invasive method to deliver light deep inside the brain tissue for neurostimulation applications which are not feasible using conventional techniques because of the high level of scattering in most tissue samples. In the developed DOPC system, the performance of the system in focusing light through and inside scattering mediums was quantified. We also showed how misalignments and imperfections of the optical components can immensely reduce the capability of a DOPC setup. Then, a systematic calibration algorithm was proposed and experimentally applied to our DOPC system to compensate main aberrations such as reference beam aberrations and also the backplane curvature of the spatial light modulator. In a highly scattering sample, the calibration algorithm achieved up to 8 fold increase in the PBR

Digital watermarking methods for data security and authentication

Philosophiae Doctor - PhDCryptology is the study of systems that typically originate from a consideration of the ideal circumstances under which secure information exchange is to take place. It involves the study of cryptographic and other processes that might be introduced for breaking the output of such systems - cryptanalysis. This includes the introduction of formal mathematical methods for the design of a cryptosystem and for estimating its theoretical level of securit

Holographic Particle Image Velocimetry of Ink Jet Streams

Ink jet technology is a rapidly growing and diverse field of research. Ink jets are used to deliver very precise and small (picolitre) volumes of fluid to a surface. Recent advances in ink jet technology demand a better understanding of the dynamics of the fluid during jetting. The aim of this project was to design a method capable of measuring the flow velocities inside ink jet streams. This objective has been achieved by the use of digital holographic particle image velocimetry. The difficulty with measuring flows inside tightly curved samples is that the refractive index change over the boundary leads to an optical distortion and therefore particles cannot be viewed or tracked reliably. Optical distortion is compensated for by taking advantage of the ability to replay a holographically recorded wave. The light scattered by particles is propagated numerically back through the sample’s surface, to form a three-dimensional image in which all refractions at the interface have been accounted for. Three dimensional particle fields are then analysed using custom particle detection and correlation code to extract the displacement of individual particles between exposures, which facilitates the construction of full flow profiles. Holograms were recorded with a simple off-axis holographic microscope, comprising two point sources of divergent light, formed from the same objective lens, acting as the source of illumination and reference light, respectively. Experiments were conducted on continuous ink jet streams of water issuing from a nozzle with 100 µm diameter. For a few millimetres after the nozzle exit, the jet is cylindrical, it then starts to form swells and necks; the swells continue to grow at the expense of the necks until the jet breaks up into a stream of droplets. Measurements of the stream wise component of velocity have been successful in the cylindrical parts of the jet, in swells and in necks greater than 20 µm in diameter. To my knowledge measurements of particle velocities on fluid jets at this scale have not been accomplished previously

Three-dimensional geometry characterization using structured light fields

Tese de doutoramento. Engenharia Mecânica. Faculdade de Engenharia. Universidade do Porto. 200

Objects extraction and recognition for camera-based interaction : heuristic and statistical approaches

In this thesis, heuristic and probabilistic methods are applied to a number of problems for camera-based interactions. The goal is to provide solutions for a vision based system that is able to extract and analyze interested objects in camera images and to use that information for various interactions for mobile usage. New methods and new attempts of combination of existing methods are developed for different applications, including text extraction from complex scene images, bar code reading performed by camera phones, and face/facial feature detection and facial expression manipulation. The application-driven problems of camera-based interaction can not be modeled by a uniform and straightforward model that has very strong simplifications of reality. The solutions we learned to be efficient were to apply heuristic but easy of implementation approaches at first to reduce the complexity of the problems and search for possible means, then use developed statistical learning approaches to deal with the remaining difficult but well-defined problems and get much better accuracy. The process can be evolved in some or all of the stages, and the combination of the approaches is problem-dependent. Contribution of this thesis resides in two aspects: firstly, new features and approaches are proposed either as heuristics or statistical means for concrete applications; secondly engineering design combining seveal methods for system optimization is studied. Geometrical characteristics and the alignment of text, texture features of bar codes, and structures of faces can all be extracted as heuristics for object extraction and further recognition. The boosting algorithm is one of the proper choices to perform probabilistic learning and to achieve desired accuracy. New feature selection techniques are proposed for constructing the weak learner and applying the boosting output in concrete applications. Subspace methods such as manifold learning algorithms are introduced and tailored for facial expression analysis and synthesis. A modified generalized learning vector quantization method is proposed to deal with the blurring of bar code images. Efficient implementations that combine the approaches in a rational joint point are presented and the results are illustrated.reviewe