Detecting low intensity nuclei on propidium iodide stained digital slides

Digitization in pathology and cytology labs is now widespread, a significant shift from a decade ago when few doctors used image processing tools. Despite unchanged scanning times due to excitation in fluorescent imaging, advancements in computing power and software have enabled more complex algorithms, yielding better-quality results. This study evaluates three nucleus segmentation algorithms for ploidy analysis using propidium iodide-stained digital WSI slides. Our goal was to improve segmentation accuracy to more closely match DNA histograms obtained via flow cytometry, with the ultimate aim of enhancing the calibration method we proposed in a previous study, which seeks to align image cytometry results with those from flow cytometry. We assessed these algorithms based on raw segmentation performance and DNA histogram similarity, using confusion-matrix-based metrics. Results indicate that modern algorithms perform better, with F1 scores exceeding 0.845, compared to our earlier solution’s 0.807, and produce DNA histograms that more closely resemble those from the reference FCM method.

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Calibration-Aimed Comparison of Image-Cytometry- and Flow-Cytometry-Based Approaches of Ploidy Analysis

Jónás¹,

Paulik²,

Kozlovszky

et al. 2022

Sensors

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Ploidy analysis is the fundamental method of measuring DNA content. For decades, the principal way of conducting ploidy analysis was through flow cytometry. A flow cytometer is a specialized tool for analyzing cells in a solution. This is convenient in laboratory environments, but prohibits measurement reproducibility and the complete detachment of sample preparation from data acquisition and analysis, which seems to have become paramount with the constant decrease in the number of pathologists per capita all over the globe. As more open computer-aided systems emerge in medicine, the demand for overcoming these shortcomings, and opening access to even more (and more flexible) options, has also emerged. Image-based analysis systems can provide an alternative to these types of workloads, placing the abovementioned problems in a different light. Flow cytometry data can be used as a reference for calibrating an image-based system. This article aims to show an approach to constructing an image-based solution for ploidy analysis, take measurements for a basic comparison of the data produced by the two methods, and produce a workflow with the ultimate goal of calibrating the image-based system.

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Detecting low intensity nuclei on propidium iodide stained digital slides

Cited by 3 publications

References 10 publications