NuSeT: A deep learning tool for reliably separating and analyzing crowded cells.

Journal: PLoS computational biology
Published Date:

Abstract

Segmenting cell nuclei within microscopy images is a ubiquitous task in biological research and clinical applications. Unfortunately, segmenting low-contrast overlapping objects that may be tightly packed is a major bottleneck in standard deep learning-based models. We report a Nuclear Segmentation Tool (NuSeT) based on deep learning that accurately segments nuclei across multiple types of fluorescence imaging data. Using a hybrid network consisting of U-Net and Region Proposal Networks (RPN), followed by a watershed step, we have achieved superior performance in detecting and delineating nuclear boundaries in 2D and 3D images of varying complexities. By using foreground normalization and additional training on synthetic images containing non-cellular artifacts, NuSeT improves nuclear detection and reduces false positives. NuSeT addresses common challenges in nuclear segmentation such as variability in nuclear signal and shape, limited training sample size, and sample preparation artifacts. Compared to other segmentation models, NuSeT consistently fares better in generating accurate segmentation masks and assigning boundaries for touching nuclei.

Authors

  • Linfeng Yang
    Bioengineering, Stanford University, Stanford, CA, United States of America.
  • Rajarshi P Ghosh
    Bioengineering, Stanford University, Stanford, CA, United States of America.
  • J Matthew Franklin
    Bioengineering, Stanford University, Stanford, CA, United States of America.
  • Simon Chen
    Department of Pathology, Stanford University School of Medicine, Stanford, CA, United States of America.
  • Chenyu You
  • Raja R Narayan
    Department of Surgery, Mass General, Boston MA, United States.
  • Marc L Melcher
    Department of Surgery, Stanford University, Stanford, CA, United States.
  • Jan T Liphardt
    Bioengineering, Stanford University, Stanford, CA, United States of America.