Abstract

Astrocytes are involved in various brain pathologies including trauma, stroke, neurodegenerative disorders such as Alzheimer’s and Parkinson’s diseases, or chronic pain. Determining cell density in a complex tissue environment in microscopy images and elucidating the temporal characteristics of morphological and biochemical changes is essential to understand the role of astrocytes in physiological and pathological conditions. Nowadays, manual stereological cell counting or semi-automatic segmentation techniques are widely used for the quantitative analysis of microscopy images. Detecting astrocytes automatically is a highly challenging computational task, for which we currently lack efficient image analysis tools. We have developed a fast and fully automated software that assesses the number of astrocytes using Deep Convolutional Neural Networks (DCNN). The method highly outperforms state-of-the-art image analysis and machine learning methods and provides precision comparable to those of human experts. Additionally, the runtime of cell detection is significantly less than that of other three computational methods analysed, and it is faster than human observers by orders of magnitude. We applied our DCNN-based method to examine the number of astrocytes in different brain regions of rats with opioid-induced hyperalgesia/tolerance (OIH/OIT), as morphine tolerance is believed to activate glia. We have demonstrated a strong positive correlation between manual and DCNN-based quantification of astrocytes in rat brain.

Highlights

  • Astrocytes are a type of glial cells within the central nervous system (CNS)

  • In this paper we introduced a novel, fully automated software FindMyCells for the accurate detection of astrocytes, a special type of neurons affected in various CNS pathologies, including trauma, stroke, neurodegenerative disorders or chronic pain

  • Astrocytes are characterized by a huge variation in appearance, size and shape which complicates their detection, their precise and high-scale identification and analysis would be essential for a better understanding of their role in the aforementioned brain pathologies

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Summary

Introduction

Astrocytes are a type of glial cells within the central nervous system (CNS). On average, there are 67–86 billion neurons and 40–85 billion glial cells in human brain[1]. Used open-source tools for general cell quantification such as ImageJ5, custom scripts or ilastik[6] have technical limitations concerning the accurate detection of astrocytes because of their complex morphology[2,7,8]. These tools are based on either machine-learning methods (ilastik) or thresholding (ImageJ, custom scripts). To validate the proposed software, we compared the cell counts produced by human experts and FindMyCells, as well as by three other software tools (ilastik, custom threshold-based script[21], ImageJ), analysing brain tissues of rats treated with repeated injections of morphine to induce opioid-induced hyperalgesia/tolerance (OIH/OIT). We analysed the microscopy images of these samples by both manual counting and FindMyCells, separately, and assessed the correlation between FindMyCell’s output and the manual counting of astrocytes

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