An end-to-end pipeline based on open source deep learning tools for reliable analysis of complex 3D images of ovaries.

Journal: Development (Cambridge, England)
PMID:

Abstract

Computational analysis of bio-images by deep learning (DL) algorithms has made exceptional progress in recent years and has become much more accessible to non-specialists with the development of ready-to-use tools. The study of oogenesis mechanisms and female reproductive success has also recently benefited from the development of efficient protocols for three-dimensional (3D) imaging of ovaries. Such datasets have a great potential for generating new quantitative data but are, however, complex to analyze due to the lack of efficient workflows for 3D image analysis. Here, we have integrated two existing open-source DL tools, Noise2Void and Cellpose, into an analysis pipeline dedicated to 3D follicular content analysis, which is available on Fiji. Our pipeline was developed on larvae and adult medaka ovaries but was also successfully applied to different types of ovaries (trout, zebrafish and mouse). Image enhancement, Cellpose segmentation and post-processing of labels enabled automatic and accurate quantification of these 3D images, which exhibited irregular fluorescent staining, low autofluorescence signal or heterogeneous follicles sizes. In the future, this pipeline will be useful for extensive cellular phenotyping in fish or mammals for developmental or toxicology studies.

Authors

  • Manon Lesage
    INRAE, Fish Physiology and Genomics Institute, 16 Allee Henri Fabre, Rennes 35000, France.
  • Manon Thomas
    INRAE, Fish Physiology and Genomics Institute, 16 Allee Henri Fabre, Rennes 35000, France.
  • Thierry Pécot
    Department of Biochemistry and Molecular Biology, Hollings Cancer Center, Medical University of South Carolina, Charleston, SC, 29407, USA.
  • Tu-Ky Ly
    INERIS, UMR-I 02 SEBIO, Verneuil en Halatte 65550, France.
  • Nathalie Hinfray
    INERIS, UMR-I 02 SEBIO, Verneuil en Halatte 65550, France.
  • Remy Beaudouin
    INERIS, UMR-I 02 SEBIO, Verneuil en Halatte 65550, France.
  • Michelle Neumann
    The Francis Crick Institute, 1 Midland Rd, London NW1 1AT, UK.
  • Robin Lovell-Badge
    The Francis Crick Institute, 1 Midland Rd, London NW1 1AT, UK.
  • Jérôme Bugeon
    INRAE, Fish Physiology and Genomics Institute, 16 Allee Henri Fabre, Rennes 35000, France.
  • Violette Thermes
    INRAE, Fish Physiology and Genomics Institute, 16 Allee Henri Fabre, Rennes 35000, France.