A robust method for high-precision quantification of the complex three-dimensional vasculatures acquired by X-ray microtomography.

The quantification of micro-vasculatures is important for the analysis of angiogenesis on which the detection of tumor growth or hepatic fibrosis depends. Synchrotron-based X-ray computed micro-tomography (SR-µCT) allows rapid acquisition of micro-vasculature images at micrometer-scale spatial resolution. Through skeletonization, the statistical features of the micro-vasculature can be extracted from the skeleton of the micro-vasculatures. Thinning is a widely used algorithm to produce the vascular skeleton in medical research. Existing three-dimensional thinning methods normally emphasize the preservation of topological structure rather than geometrical features in generating the skeleton of a volumetric object. This results in three problems and limits the accuracy of the quantitative results related to the geometrical structure of the vasculature. The problems include the excessively shortened length of elongated objects, eliminated branches of blood vessel tree structure, and numerous noisy spurious branches. The inaccuracy of the skeleton directly introduces errors in the quantitative analysis, especially on the parameters concerning the vascular length and the counts of vessel segments and branching points. In this paper, a robust method using a consolidated end-point constraint for thinning, which generates geometry-preserving skeletons in addition to maintaining the topology of the vasculature, is presented. The improved skeleton can be used to produce more accurate quantitative results. Experimental results from high-resolution SR-µCT images show that the end-point constraint produced by the proposed method can significantly improve the accuracy of the skeleton obtained using the existing ITK three-dimensional thinning filter. The produced skeleton has laid the groundwork for accurate quantification of the angiogenesis. This is critical for the early detection of tumors and assessing anti-angiogenesis treatments.

Towards automated quantitative vasculature understanding via ultra high-resolution imagery.

This chapter presents an approach to processing ultra high-resolution, large-size biomedical imaging data for the purposes of detecting and quantifying vasculature and microvasculature . Capturing early signs of any changes in vasculature may have significant values for early-diagnosis and treatment assessment due to the well understood observation that vascular changes precede cancerous growth and metastasis metastasis . With the advent of key enabling technologies for extremely high-resolution imaging, such as synchrotron radiation synchrotron radiation based computed tomography (CT) computed tomography , the required levels of detail have become accessible. However, these technologies also present challenges in data analysis. This chapter aims to offer some insights as to how these changes might be best dealt with. We argue that the necessary steps in quantitative understanding of vasculatures include targeted data enhancement enhancement , information reduction aimed at characterizing the linear structure of vessels vessels , and quantitatively describing the vessel hierarchy. We present results on cerebral and liver vasculatures of a mouse captured at the Shanghai Synchrotron Radiation Facility (SSRF). These results were achieved with a processing pipeline comprising of our empirically selected component for each of the above steps. Towards the end, we discuss how alternative and additional components may be incorporated for improved speed and robustness.

Pollen image classification using the Classifynder system: algorithm comparison and a case study on New Zealand honey.

We describe an investigation into how Massey University's Pollen Classifynder can accelerate the understanding of pollen and its role in nature. The Classifynder is an imaging microscopy system that can locate, image and classify slide based pollen samples. Given the laboriousness of purely manual image acquisition and identification it is vital to exploit assistive technologies like the Classifynder to enable acquisition and analysis of pollen samples. It is also vital that we understand the strengths and limitations of automated systems so that they can be used (and improved) to compliment the strengths and weaknesses of human analysts to the greatest extent possible. This article reviews some of our experiences with the Classifynder system and our exploration of alternative classifier models to enhance both accuracy and interpretability. Our experiments in the pollen analysis problem domain have been based on samples from the Australian National University's pollen reference collection (2,890 grains, 15 species) and images bundled with the Classifynder system (400 grains, 4 species). These samples have been represented using the Classifynder image feature set. We additionally work through a real world case study where we assess the ability of the system to determine the pollen make-up of samples of New Zealand honey. In addition to the Classifynder's native neural network classifier, we have evaluated linear discriminant, support vector machine, decision tree and random forest classifiers on these data with encouraging results. Our hope is that our findings will help enhance the performance of future releases of the Classifynder and other systems for accelerating the acquisition and analysis of pollen samples.

Cloud based toolbox for image analysis, processing and reconstruction tasks.

This chapter describes a novel way of carrying out image analysis, reconstruction and processing tasks using cloud based service provided on the Australian National eResearch Collaboration Tools and Resources (NeCTAR) infrastructure. The toolbox allows users free access to a wide range of useful blocks of functionalities (imaging functions) that can be connected together in workflows allowing creation of even more complex algorithms that can be re-run on different data sets, shared with others or additionally adjusted. The functions given are in the area of cellular imaging, advanced X-ray image analysis, computed tomography and 3D medical imaging and visualisation. The service is currently available on the website www.cloudimaging.net.au .

HCA-vision: Automated neurite outgrowth analysis.

Automating the analysis of neurons in culture represents a key aspect of the search for neuroactive compounds. A number of commercial neurite analysis software packages tend to measure some basic features such as total neurite length and number of branching points. However, with only these measurements, some differences between neurite morphologies that are clear to a human observer cannot be identified. The authors have developed a suite of image analysis tools that will allow researchers to produce quality analyses at primary screening rates. The suite provides sensitive and information-rich measurements of neurons and neurites. It can discriminate subtle changes in complex neurite arborization even when neurons and neurites are dense. This allows users to selectively screen for compounds triggering different types of neurite outgrowth behavior. In mixed cell populations, neurons can be filtered and separated from other brain cell types so that neurite analysis can be performed only on neurons. It supports batch processing with a built-in database to store the batch-processing results, a batch result viewer, and an ad hoc query builder for users to retrieve features of interest. The suite of tools has been deployed into a software package called HCA-Vision. The free version of the software package is available at http://www.hca-vision.com.

Automated analysis of neurite branching in cultured cortical neurons using HCA-Vision.

Manual neuron tracing is a very labor-intensive task. In the drug screening context, the sheer number of images to process means that this approach is unrealistic. Moreover, the lack of reproducibility, objectivity, and auditing capability of manual tracing is limiting even in the context of smaller studies. We have developed fast, sensitive, and reliable algorithms for the purpose of detecting and analyzing neurites in cell cultures, and we have integrated them in software called HCA-Vision, suitable for the research environment. We validate the software on images of cortical neurons by comparing results obtained using HCA-Vision with those obtained using an established semi-automated tracing solution (NeuronJ). The effect of the Sez-6 deletion was characterized in detail. Sez-6 null neurons exhibited a significant increase in neurite branching, although the neurite field area was unchanged due to a reduction in mean branch length. HCA-Vision delivered considerable speed benefits and reliable traces.