Sensitivity of edge detection methods for quantifying cell migration assays

Quantitative imaging methods to analyze cell migration assays are not standardized. Here we present a suite of two–dimensional barrier assays describing the collective spreading of an initially–confined population of 3T3 fibroblast cells. To quantify the motility rate we apply two different automatic image detection methods to locate the position of the leading edge of the spreading population after 24, 48 and 72 hours. These results are compared with a manual edge detection method where we systematically vary the detection threshold. Our results indicate that the observed spreading rates are very sensitive to the choice of image analysis tools and we show that a standard measure of cell migration can vary by as much as 25% for the same experimental images depending on the details of the image analysis tools. Our results imply that it is very difficult, if not impossible, to meaningfully compare previously published measures of cell migration since previous results have been obtained using different image analysis techniques and the details of these techniques are not always reported. Using a mathematical model, we provide a physical interpretation of our edge detection results. The physical interpretation is important since edge detection algorithms alone do not specify any physical measure, or physical definition, of the leading edge of the spreading population. Our modeling indicates that variations in the image threshold parameter correspond to a consistent variation in the local cell density. This means that varying the threshold parameter is equivalent to varying the location of the leading edge in the range of approximately 1–5% of the maximum cell density.

vSpeak: Edge detection based feature extraction for sign to text conversion

This paper presents 'vSpeak', the first initiative taken in Pakistan for ICT enabled conversion of dynamic Sign Urdu gestures into natural language sentences. To realize this, vSpeak has adopted a novel approach for feature extraction using edge detection and image compression which gives input to the Artificial Neural Network that recognizes the gesture. This technique caters for the blurred images as well. The training and testing is currently being performed on a dataset of 200 patterns of 20 words from Sign Urdu with target accuracy of 90% and above.

Breast cancer detection from thermal images using bispectral invariant features

Highly sensitive infrared (IR) cameras provide high-resolution diagnostic images of the temperature and vascular changes of breasts. These images can be processed to emphasize hot spots that exhibit early and subtle changes owing to pathology. The resulting images show clusters that appear random in shape and spatial distribution but carry class dependent information in shape and texture. Automated pattern recognition techniques are challenged because of changes in location, size and orientation of these clusters. Higher order spectral invariant features provide robustness to such transformations and are suited for texture and shape dependent information extraction from noisy images. In this work, the effectiveness of bispectral invariant features in diagnostic classification of breast thermal images into malignant, benign and normal classes is evaluated and a phase-only variant of these features is proposed. High resolution IR images of breasts, captured with measuring accuracy of ±0.4% (full scale) and temperature resolution of 0.1 °C black body, depicting malignant, benign and normal pathologies are used in this study. Breast images are registered using their lower boundaries, automatically extracted using landmark points whose locations are learned during training. Boundaries are extracted using Canny edge detection and elimination of inner edges. Breast images are then segmented using fuzzy c-means clustering and the hottest regions are selected for feature extraction. Bispectral invariant features are extracted from Radon projections of these images. An Adaboost classifier is used to select and fuse the best features during training and then classify unseen test images into malignant, benign and normal classes. A data set comprising 9 malignant, 12 benign and 11 normal cases is used for evaluation of performance. Malignant cases are detected with 95% accuracy. A variant of the features using the normalized bispectrum, which discards all magnitude information, is shown to perform better for classification between benign and normal cases, with 83% accuracy compared to 66% for the original.

Analysis of micro-structural changes and measurement of their parameters of a food material

Food microstructure represents the way their elements arrangement and their interaction. Researchers in this field benefit from identifying new methods of examination of the microstructure and analysing the images. Experiments were undertaken to study micro-structural changes of food material during drying. Micro-structural images were obtained for potato samples of cubical shape at different moisture contents during drying using scanning electron microscopy. Physical parameters such as cell wall perimeter, and area were calculated using an image identification algorithm, based on edge detection and morphological operators. The algorithm was developed using Matlab.

SwarmPS: Rapid, semi-automated single particle selection software

Single particle analysis (SPA) coupled with high-resolution electron cryo-microscopy is emerging as a powerful technique for the structure determination of membrane protein complexes and soluble macromolecular assemblies. Current estimates suggest that ∼104–105 particle projections are required to attain a 3 Å resolution 3D reconstruction (symmetry dependent). Selecting this number of molecular projections differing in size, shape and symmetry is a rate-limiting step for the automation of 3D image reconstruction. Here, we present SwarmPS, a feature rich GUI based software package to manage large scale, semi-automated particle picking projects. The software provides cross-correlation and edge-detection algorithms. Algorithm-specific parameters are transparently and automatically determined through user interaction with the image, rather than by trial and error. Other features include multiple image handling (∼102), local and global particle selection options, interactive image freezing, automatic particle centering, and full manual override to correct false positives and negatives. SwarmPS is user friendly, flexible, extensible, fast, and capable of exporting boxed out projection images, or particle coordinates, compatible with downstream image processing suites.

Asphalt concrete surfaces macrotexture determination from still images

Road surface macrotexture is identified as one of the factors contributing to the surface's skid resistance. Existing methods of quantifying the surface macrotexture, such as the sand patch test and the laser profilometer test, are either expensive or intrusive, requiring traffic control. High-resolution cameras have made it possible to acquire good quality images from roads for the automated analysis of texture depth. In this paper, a granulometric method based on image processing is proposed to estimate road surface texture coarseness distribution from their edge profiles. More than 1300 images were acquired from two different sites, extending to a total of 2.96 km. The images were acquired using camera orientations of 60 and 90 degrees. The road surface is modeled as a texture of particles, and the size distribution of these particles is obtained from chord lengths across edge boundaries. The mean size from each distribution is compared with the sensor measured texture depth obtained using a laser profilometer. By tuning the edge detector parameters, a coefficient of determination of up to R2 = 0.94 between the proposed method and the laser profilometer method was obtained. The high correlation is also confirmed by robust calibration parameters that enable the method to be used for unseen data after the method has been calibrated over road surface data with similar surface characteristics and under similar imaging conditions.

Particulate texture image analysis with applications

Texture analysis and textural cues have been applied for image classification, segmentation and pattern recognition. Dominant texture descriptors include directionality, coarseness, line-likeness etc. In this dissertation a class of textures known as particulate textures are defined, which are predominantly coarse or blob-like. The set of features that characterise particulate textures are different from those that characterise classical textures. These features are micro-texture, macro-texture, size, shape and compaction. Classical texture analysis techniques do not adequately capture particulate texture features. This gap is identified and new methods for analysing particulate textures are proposed. The levels of complexity in particulate textures are also presented ranging from the simplest images where blob-like particles are easily isolated from their back- ground to the more complex images where the particles and the background are not easily separable or the particles are occluded. Simple particulate images can be analysed for particle shapes and sizes. Complex particulate texture images, on the other hand, often permit only the estimation of particle dimensions. Real life applications of particulate textures are reviewed, including applications to sedimentology, granulometry and road surface texture analysis. A new framework for computation of particulate shape is proposed. A granulometric approach for particle size estimation based on edge detection is developed which can be adapted to the gray level of the images by varying its parameters. This study binds visual texture analysis and road surface macrotexture in a theoretical framework, thus making it possible to apply monocular imaging techniques to road surface texture analysis. Results from the application of the developed algorithm to road surface macro-texture, are compared with results based on Fourier spectra, the auto- correlation function and wavelet decomposition, indicating the superior performance of the proposed technique. The influence of image acquisition conditions such as illumination and camera angle on the results was systematically analysed. Experimental data was collected from over 5km of road in Brisbane and the estimated coarseness along the road was compared with laser profilometer measurements. Coefficient of determination R2 exceeding 0.9 was obtained when correlating the proposed imaging technique with the state of the art Sensor Measured Texture Depth (SMTD) obtained using laser profilometers.

Graph rigidity for near-coplanar structure from motion

Recent algorithms for monocular motion capture (MoCap) estimate weak-perspective camera matrices between images using a small subset of approximately-rigid points on the human body (i.e. the torso and hip). A problem with this approach, however, is that these points are often close to coplanar, causing canonical linear factorisation algorithms for rigid structure from motion (SFM) to become extremely sensitive to noise. In this paper, we propose an alternative solution to weak-perspective SFM based on a convex relaxation of graph rigidity. We demonstrate the success of our algorithm on both synthetic and real world data, allowing for much improved solutions to marker less MoCap problems on human bodies. Finally, we propose an approach to solve the two-fold ambiguity over bone direction using a k-nearest neighbour kernel density estimator.

3D reconstruction of long bones utilising magnetic resonance imaging (MRI)

The design of pre-contoured fracture fixation implants (plates and nails) that correctly fit the anatomy of a patient utilises 3D models of long bones with accurate geometric representation. 3D data is usually available from computed tomography (CT) scans of human cadavers that generally represent the above 60 year old age group. Thus, despite the fact that half of the seriously injured population comes from the 30 year age group and below, virtually no data exists from these younger age groups to inform the design of implants that optimally fit patients from these groups. Hence, relevant bone data from these age groups is required. The current gold standard for acquiring such data–CT–involves ionising radiation and cannot be used to scan healthy human volunteers. Magnetic resonance imaging (MRI) has been shown to be a potential alternative in the previous studies conducted using small bones (tarsal bones) and parts of the long bones. However, in order to use MRI effectively for 3D reconstruction of human long bones, further validations using long bones and appropriate reference standards are required. Accurate reconstruction of 3D models from CT or MRI data sets requires an accurate image segmentation method. Currently available sophisticated segmentation methods involve complex programming and mathematics that researchers are not trained to perform. Therefore, an accurate but relatively simple segmentation method is required for segmentation of CT and MRI data. Furthermore, some of the limitations of 1.5T MRI such as very long scanning times and poor contrast in articular regions can potentially be reduced by using higher field 3T MRI imaging. However, a quantification of the signal to noise ratio (SNR) gain at the bone - soft tissue interface should be performed; this is not reported in the literature. As MRI scanning of long bones has very long scanning times, the acquired images are more prone to motion artefacts due to random movements of the subject‟s limbs. One of the artefacts observed is the step artefact that is believed to occur from the random movements of the volunteer during a scan. This needs to be corrected before the models can be used for implant design. As the first aim, this study investigated two segmentation methods: intensity thresholding and Canny edge detection as accurate but simple segmentation methods for segmentation of MRI and CT data. The second aim was to investigate the usability of MRI as a radiation free imaging alternative to CT for reconstruction of 3D models of long bones. The third aim was to use 3T MRI to improve the poor contrast in articular regions and long scanning times of current MRI. The fourth and final aim was to minimise the step artefact using 3D modelling techniques. The segmentation methods were investigated using CT scans of five ovine femora. The single level thresholding was performed using a visually selected threshold level to segment the complete femur. For multilevel thresholding, multiple threshold levels calculated from the threshold selection method were used for the proximal, diaphyseal and distal regions of the femur. Canny edge detection was used by delineating the outer and inner contour of 2D images and then combining them to generate the 3D model. Models generated from these methods were compared to the reference standard generated using the mechanical contact scans of the denuded bone. The second aim was achieved using CT and MRI scans of five ovine femora and segmenting them using the multilevel threshold method. A surface geometric comparison was conducted between CT based, MRI based and reference models. To quantitatively compare the 1.5T images to the 3T MRI images, the right lower limbs of five healthy volunteers were scanned using scanners from the same manufacturer. The images obtained using the identical protocols were compared by means of SNR and contrast to noise ratio (CNR) of muscle, bone marrow and bone. In order to correct the step artefact in the final 3D models, the step was simulated in five ovine femora scanned with a 3T MRI scanner. The step was corrected using the iterative closest point (ICP) algorithm based aligning method. The present study demonstrated that the multi-threshold approach in combination with the threshold selection method can generate 3D models from long bones with an average deviation of 0.18 mm. The same was 0.24 mm of the single threshold method. There was a significant statistical difference between the accuracy of models generated by the two methods. In comparison, the Canny edge detection method generated average deviation of 0.20 mm. MRI based models exhibited 0.23 mm average deviation in comparison to the 0.18 mm average deviation of CT based models. The differences were not statistically significant. 3T MRI improved the contrast in the bone–muscle interfaces of most anatomical regions of femora and tibiae, potentially improving the inaccuracies conferred by poor contrast of the articular regions. Using the robust ICP algorithm to align the 3D surfaces, the step artefact that occurred by the volunteer moving the leg was corrected, generating errors of 0.32 ± 0.02 mm when compared with the reference standard. The study concludes that magnetic resonance imaging, together with simple multilevel thresholding segmentation, is able to produce 3D models of long bones with accurate geometric representations. The method is, therefore, a potential alternative to the current gold standard CT imaging.

Mawson the astrobiologist rover : towards automatic recognition of stromatolites

This work aims at developing a planetary rover capable of acting as an assistant astrobiologist: making a preliminary analysis of the collected visual images that will help to make better use of the scientists time by pointing out the most interesting pieces of data. This paper focuses on the problem of detecting and recognising particular types of stromatolites. Inspired by the processes actual astrobiologists go through in the field when identifying stromatolites, the processes we investigate focus on recognising characteristics associated with biogenicity. The extraction of these characteristics is based on the analysis of geometrical structure enhanced by passing the images of stromatolites into an edge-detection filter and its Fourier Transform, revealing typical spatial frequency patterns. The proposed analysis is performed on both simulated images of stromatolite structures and images of real stromatolites taken in the field by astrobiologists.