Abstract

Advances in microscopy imaging technologies have enabled the visualization of live-cell dynamic processes using time-lapse microscopy imaging. However, modern methods exhibit several limitations related to the training phases and to time constraints, hindering their application in the laboratory practice. In this work, we present a novel method, named Automated Cell Detection and Counting (ACDC), designed for activity detection of fluorescent labeled cell nuclei in time-lapse microscopy. ACDC overcomes the limitations of the literature methods, by first applying bilateral filtering on the original image to smooth the input cell images while preserving edge sharpness, and then by exploiting the watershed transform and morphological filtering. Moreover, ACDC represents a feasible solution for the laboratory practice, as it can leverage multi-core architectures in computer clusters to efficiently handle large-scale imaging datasets. Indeed, our Parent-Workers implementation of ACDC allows to obtain up to a 3.7× speed-up compared to the sequential counterpart. ACDC was tested on two distinct cell imaging datasets to assess its accuracy and effectiveness on images with different characteristics. We achieved an accurate cell-count and nuclei segmentation without relying on large-scale annotated datasets, a result confirmed by the average Dice Similarity Coefficients of 76.84 and 88.64 and the Pearson coefficients of 0.99 and 0.96, calculated against the manual cell counting, on the two tested datasets.

Highlights

  • Advances in microscopy imaging technologies have enabled the visualization of dynamic live-cell processes using time-lapse microscopy methodologies [1,2,3]

  • With the goal of validating ACDC on imaging data coming from different sources, we considered a selection of the training set of the Data Science Bowl (DSB) dataset [37,46], which was a competition organized by Kaggle (San Francisco, CA, USA)

  • For the 2018 DSB dataset (Figure 8b), ACDC is capable of accurately separating groups of cells that were erroneously delineated as single connected-components in the gold standard; the spurious speckles included in the 2018 DSB gold standard, are not detected by ACDC since these very small regions are characterized by hypo-intense fluorescence levels

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Summary

Introduction

Advances in microscopy imaging technologies have enabled the visualization of dynamic live-cell processes using time-lapse microscopy methodologies [1,2,3]. The objects of interest in cellular images are characterized by high variations of morphology and intensity from image to image [6], which make features such as cell boundaries and intracellular features difficult to accurately identify For this reason, cell segmentation analysis has gained increasing attention over the last decade [7]. The most commonly used free and open-source software tools for microscopy applications in the laboratory are ImageJ [8] or Fiji [9], and CellProfiler [10]. These tools offer customization capabilities, they do not provide suitable functionalities for fast and efficient high-throughput cell image analysis on large-scale datasets. CellProfiler Analyst [11] allows the user to explore and visualize image-based data and to classify complex biological phenotypes with classic supervised

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