Dataset Information

Automatic Detection of Freshwater Phytoplankton Specimens in Conventional Microscopy Images.

ABSTRACT: Water safety and quality can be compromised by the proliferation of toxin-producing phytoplankton species, requiring continuous monitoring of water sources. This analysis involves the identification and counting of these species which requires broad experience and knowledge. The automatization of these tasks is highly desirable as it would release the experts from tedious work, eliminate subjective factors, and improve repeatability. Thus, in this preliminary work, we propose to advance towards an automatic methodology for phytoplankton analysis in digital images of water samples acquired using regular microscopes. In particular, we propose a novel and fully automatic method to detect and segment the existent phytoplankton specimens in these images using classical computer vision algorithms. The proposed method is able to correctly detect sparse colonies as single phytoplankton candidates, thanks to a novel fusion algorithm, and is able to differentiate phytoplankton specimens from other image objects in the microscope samples (like minerals, bubbles or detritus) using a machine learning based approach that exploits texture and colour features. Our preliminary experiments demonstrate that the proposed method provides satisfactory and accurate results.

SUBMITTER: Rivas-Villar D

PROVIDER: S-EPMC7700267 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature

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Publications

Automatic Detection of Freshwater Phytoplankton Specimens in Conventional Microscopy Images.

Rivas-Villar David D Rouco José J Penedo Manuel G MG Carballeira Rafael R Novo Jorge J

Sensors (Basel, Switzerland) 20201123 22

Water safety and quality can be compromised by the proliferation of toxin-producing phytoplankton species, requiring continuous monitoring of water sources. This analysis involves the identification and counting of these species which requires broad experience and knowledge. The automatization of these tasks is highly desirable as it would release the experts from tedious work, eliminate subjective factors, and improve repeatability. Thus, in this preliminary work, we propose to advance towards ...[more]

PMID: 33238566

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Project description:BackgroundIn recent years, high-throughput microscopy has emerged as a powerful tool to analyze cellular dynamics in an unprecedentedly high resolved manner. The amount of data that is generated, for example in long-term time-lapse microscopy experiments, requires automated methods for processing and analysis. Available software frameworks are well suited for high-throughput processing of fluorescence images, but they often do not perform well on bright field image data that varies considerably between laboratories, setups, and even single experiments.ResultsIn this contribution, we present a fully automated image processing pipeline that is able to robustly segment and analyze cells with ellipsoid morphology from bright field microscopy in a high-throughput, yet time efficient manner. The pipeline comprises two steps: (i) Image acquisition is adjusted to obtain optimal bright field image quality for automatic processing. (ii) A concatenation of fast performing image processing algorithms robustly identifies single cells in each image. We applied the method to a time-lapse movie consisting of ∼315,000 images of differentiating hematopoietic stem cells over 6 days. We evaluated the accuracy of our method by comparing the number of identified cells with manual counts. Our method is able to segment images with varying cell density and different cell types without parameter adjustment and clearly outperforms a standard approach. By computing population doubling times, we were able to identify three growth phases in the stem cell population throughout the whole movie, and validated our result with cell cycle times from single cell tracking.ConclusionsOur method allows fully automated processing and analysis of high-throughput bright field microscopy data. The robustness of cell detection and fast computation time will support the analysis of high-content screening experiments, on-line analysis of time-lapse experiments as well as development of methods to automatically track single-cell genealogies.

Project description:Accurate identification of cell nuclei and their tracking using three dimensional (3D) microscopic images is a demanding task in many biological studies. Manual identification of nuclei centroids from images is an error-prone task, sometimes impossible to accomplish due to low contrast and the presence of noise. Nonetheless, only a few methods are available for 3D bioimaging applications, which sharply contrast with 2D analysis, where many methods already exist. In addition, most methods essentially adopt segmentation for which a reliable solution is still unknown, especially for 3D bio-images having juxtaposed cells. In this work, we propose a new method that can directly extract nuclei centroids from fluorescence microscopy images. This method involves three steps: (i) Pre-processing, (ii) Local enhancement, and (iii) Centroid extraction. The first step includes two variations: first variation (Variant-1) uses the whole 3D pre-processed image, whereas the second one (Variant-2) modifies the preprocessed image to the candidate regions or the candidate hybrid image for further processing. At the second step, a multiscale cube filtering is employed in order to locally enhance the pre-processed image. Centroid extraction in the third step consists of three stages. In Stage-1, we compute a local characteristic ratio at every voxel and extract local maxima regions as candidate centroids using a ratio threshold. Stage-2 processing removes spurious centroids from Stage-1 results by analyzing shapes of intensity profiles from the enhanced image. An iterative procedure based on the nearest neighborhood principle is then proposed to combine if there are fragmented nuclei. Both qualitative and quantitative analyses on a set of 100 images of 3D mouse embryo are performed. Investigations reveal a promising achievement of the technique presented in terms of average sensitivity and precision (i.e., 88.04% and 91.30% for Variant-1; 86.19% and 95.00% for Variant-2), when compared with an existing method (86.06% and 90.11%), originally developed for analyzing C. elegans images.

Dataset Information

Automatic Detection of Freshwater Phytoplankton Specimens in Conventional Microscopy Images.

Publications

Automatic Detection of Freshwater Phytoplankton Specimens in Conventional Microscopy Images.

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