Unsupervised segmentation of noisy electron microscopy images using salient watersheds and region merging

BACKGROUND: Segmenting electron microscopy (EM) images of cellular and subcellular processes in the nervous system is a key step in many bioimaging pipelines involving classification and labeling of ultrastructures. However, fully automated techniques to segment images are often susceptible to noise and heterogeneity in EM images (e.g. different histological preparations, different organisms, different brain regions, etc.). Supervised techniques to address this problem are often helpful but require large sets of training data, which are often difficult to obtain in practice, especially across many conditions. RESULTS: We propose a new, principled unsupervised algorithm to segment EM images using a two-step approach: edge detection via salient watersheds following by robust region merging. We performed experiments to gather EM neuroimages of two organisms (mouse and fruit fly) using different histological preparations and generated manually curated ground-truth segmentations. We compared our algorithm against several state-of-the-art unsupervised segmentation algorithms and found superior performance using two standard measures of under-and over-segmentation error. CONCLUSIONS: Our algorithm is general and may be applicable to other large-scale segmentation problems for bioimages

Large-scale automated identification of mouse brain cells in confocal light sheet microscopy images

Motivation: Recently, confocal light sheet microscopy has enabled high-throughput acquisition of whole mouse brain 3D images at the micron scale resolution. This poses the unprecedented challenge of creating accurate digital maps of the whole set of cells in a brain. Results: We introduce a fast and scalable algorithm for fully automated cell identification. We obtained the whole digital map of Purkinje cells in mouse cerebellum consisting of a set of 3D cell center coordinates. The method is accurate and we estimated an F(1) measure of 0.96 using 56 representative volumes, totaling 1.09 GVoxel and containing 4138 manually annotated soma centers. Availability and implementation: Source code and its documentation are available at http://bcfind.dinfo.unifi.it/. The whole pipeline of methods is implemented in Python and makes use of Pylearn2 and modified parts of Scikit-learn. Brain images are available on request. Contact: [email protected] Supplementary information: Supplementary data are available at Bioinformatics online

Automated analysis of colorectal cancer

Colorectal cancer (CRC) is the second largest cause of cancer deaths in the UK, with approximately 16,000 per year. Over 41,000 people are diagnosed annually, and 43% of those will die within ten years of diagnosis. The treatment of CRC patients relies on pathological examination of the disease to identify visual features that predict growth and spread, and response to chemoradiotherapy. These prognostic features are identified manually, and are subject to inter and intra-scorer variability. This variability stems from the subjectivity in interpreting large images which can have very varied appearances, as well as the time consuming and laborious methodology of visually inspecting cancer cells. The work in this thesis presents a systematic approach to developing a solution to address this problem for one such prognostic indicator, the Tumour:Stroma Ratio (TSR). The steps taken are presented sequentially through the chapters, in order of the work carried out. These specifically involve the acquisition and assessment of a dataset of 2.4 million expert-classified images of CRC, and multiple iterations of algorithm development, to automate the process of generating TSRs for patient cases. The algorithm improvements are made using conclusions from observer studies, conducted on a psychophysics experiment platform developed as part of this work, and further work is undertaken to identify issues of image quality that affect automated solutions. The developed algorithm is then applied to a clinical trial dataset with survival data, meaning that the algorithm is validated against two separate pathologist-scored, clinical trial datasets, as well as being able to test its suitability for generating independent prognostic markers