Pollen segmentation and feature evaluation for automatic classification in bright-field microscopy

14 págs.; 10 figs.; 7 tabs.; 1 app.© 2014 Elsevier B.V. Besides the well-established healthy properties of pollen, palynology and apiculture are of extreme importance to avoid hard and fast unbalances in our ecosystems. To support such disciplines computer vision comes to alleviate tedious recognition tasks. In this paper we present an applied study of the state of the art in pattern recognition techniques to describe, analyze, and classify pollen grains in an extensive dataset specifically collected (15 types, 120 samples/type). We also propose a novel contour-inner segmentation of grains, improving 50% of accuracy. In addition to published morphological, statistical, and textural descriptors, we introduce a new descriptor to measure the grain's contour profile and a logGabor implementation not tested before for this purpose. We found a significant improvement for certain combinations of descriptors, providing an overall accuracy above 99%. Finally, some palynological features that are still difficult to be integrated in computer systems are discussed.This work has been supported by the European project APIFRESH FP7-SME-2008-2 ‘‘Developing European standards for bee pollen and royal jelly: quality, safety and authenticity’’ and we would like to thank to Mr. Walter Haefeker, President of the European Professional Beekeepers Association (EPBA). J. Victor Marcos is a ‘‘Juan de la Cierva’’ research fellow funded by the Spanish Ministry of Economy and Competitiveness. Rodrigo Nava thanks Consejo Nacional de Ciencia y Tecnología (CONACYT) and PAPIIT Grant IG100814.Peer Reviewe

Hardware and software integration and testing for the automation of bright-field microscopy for tuberculosis detection

Automated microscopy for the detection of tuberculosis (TB) in sputum smears would reduce the load on technicians, especially in countries with a high TB burden. This dissertation reports on the development and testing of an automated system built around a conventional microscope for the detection of TB in Ziehl-Neelsen (ZN) stained sputum smears. Microscope auto-focusing, image analysis and stage movement were integrated. Images were captured at 40x magnification

Mathematical Morphology for Quantification in Biological & Medical Image Analysis

Mathematical morphology is an established field of image processing first introduced as an application of set and lattice theories. Originally used to characterise particle distributions, mathematical morphology has gone on to be a core tool required for such important analysis methods as skeletonisation and the watershed transform. In this thesis, I introduce a selection of new image analysis techniques based on mathematical morphology. Utilising assumptions of shape, I propose a new approach for the enhancement of vessel-like objects in images: the bowler-hat transform. Built upon morphological operations, this approach is successful at challenges such as junctions and robust against noise. The bowler-hat transform is shown to give better results than competitor methods on challenging data such as retinal/fundus imagery. Building further on morphological operations, I introduce two novel methods for particle and blob detection. The first of which is developed in the context of colocalisation, a standard biological assay, and the second, which is based on Hilbert-Edge Detection And Ranging (HEDAR), with regard to nuclei detection and counting in fluorescent microscopy. These methods are shown to produce accurate and informative results for sub-pixel and supra-pixel object counting in complex and noisy biological scenarios. I propose a new approach for the automated extraction and measurement of object thickness for intricate and complicated vessels, such as brain vascular in medical images. This pipeline depends on two key technologies: semi-automated segmentation by advanced level-set methods and automatic thickness calculation based on morphological operations. This approach is validated and results demonstrating the broad range of challenges posed by these images and the possible limitations of this pipeline are shown. This thesis represents a significant contribution to the field of image processing using mathematical morphology and the methods within are transferable to a range of complex challenges present across biomedical image analysis

Plant Seed Identification

Plant seed identification is routinely performed for seed certification in seed trade, phytosanitary certification for the import and export of agricultural commodities, and regulatory monitoring, surveillance, and enforcement. Current identification is performed manually by seed analysts with limited aiding tools. Extensive expertise and time is required, especially for small, morphologically similar seeds. Computers are, however, especially good at recognizing subtle differences that humans find difficult to perceive. In this thesis, a 2D, image-based computer-assisted approach is proposed. The size of plant seeds is extremely small compared with daily objects. The microscopic images of plant seeds are usually degraded by defocus blur due to the high magnification of the imaging equipment. It is necessary and beneficial to differentiate the in-focus and blurred regions given that only sharp regions carry distinctive information usually for identification. If the object of interest, the plant seed in this case, is in- focus under a single image frame, the amount of defocus blur can be employed as a cue to separate the object and the cluttered background. If the defocus blur is too strong to obscure the object itself, sharp regions of multiple image frames acquired at different focal distance can be merged together to make an all-in-focus image. This thesis describes a novel non-reference sharpness metric which exploits the distribution difference of uniform LBP patterns in blurred and non-blurred image regions. It runs in realtime on a single core cpu and responses much better on low contrast sharp regions than the competitor metrics. Its benefits are shown both in defocus segmentation and focal stacking. With the obtained all-in-focus seed image, a scale-wise pooling method is proposed to construct its feature representation. Since the imaging settings in lab testing are well constrained, the seed objects in the acquired image can be assumed to have measureable scale and controllable scale variance. The proposed method utilizes real pixel scale information and allows for accurate comparison of seeds across scales. By cross-validation on our high quality seed image dataset, better identification rate (95%) was achieved compared with pre- trained convolutional-neural-network-based models (93.6%). It offers an alternative method for image based identification with all-in-focus object images of limited scale variance. The very first digital seed identification tool of its kind was built and deployed for test in the seed laboratory of Canadian food inspection agency (CFIA). The proposed focal stacking algorithm was employed to create all-in-focus images, whereas scale-wise pooling feature representation was used as the image signature. Throughput, workload, and identification rate were evaluated and seed analysts reported significantly lower mental demand (p = 0.00245) when using the provided tool compared with manual identification. Although the identification rate in practical test is only around 50%, I have demonstrated common mistakes that have been made in the imaging process and possible ways to deploy the tool to improve the recognition rate

Deep-learning assisted detection and quantification of (oo)cysts of Giardia and Cryptosporidium on smartphone microscopy images

The consumption of microbial-contaminated food and water is responsible for the deaths of millions of people annually. Smartphone-based microscopy systems are portable, low-cost, and more accessible alternatives for the detection of Giardia and Cryptosporidium than traditional brightfield microscopes. However, the images from smartphone microscopes are noisier and require manual cyst identification by trained technicians, usually unavailable in resource-limited settings. Automatic detection of (oo)cysts using deep-learning-based object detection could offer a solution for this limitation. We evaluate the performance of three state-of-the-art object detectors to detect (oo)cysts of Giardia and Cryptosporidium on a custom dataset that includes both smartphone and brightfield microscopic images from vegetable samples. Faster RCNN, RetinaNet, and you only look once (YOLOv8s) deep-learning models were employed to explore their efficacy and limitations. Our results show that while the deep-learning models perform better with the brightfield microscopy image dataset than the smartphone microscopy image dataset, the smartphone microscopy predictions are still comparable to the prediction performance of non-experts.Comment: 18 pages (including supplementary information), 4 figures, 7 tables, submitting to Journal of Machine Learning for Biomedical Imagin