3D fluorescence microscopy data synthesis for segmentation and benchmarking

Automated image processing approaches are indispensable for many biomedical experiments and help to cope with the increasing amount of microscopy image data in a fast and reproducible way. Especially state-of-the-art deep learning-based approaches most often require large amounts of annotated training data to produce accurate and generalist outputs, but they are often compromised by the general lack of those annotated data sets. In this work, we propose how conditional generative adversarial networks can be utilized to generate realistic image data for 3D fluorescence microscopy from annotation masks of 3D cellular structures. In combination with mask simulation approaches, we demonstrate the generation of fully-annotated 3D microscopy data sets that we make publicly available for training or benchmarking. An additional positional conditioning of the cellular structures enables the reconstruction of position-dependent intensity characteristics and allows to generate image data of different quality levels. A patch-wise working principle and a subsequent full-size reassemble strategy is used to generate image data of arbitrary size and different organisms. We present this as a proof-of-concept for the automated generation of fully-annotated training data sets requiring only a minimum of manual interaction to alleviate the need of manual annotations.

[1]  Liyuan Liu,et al.  On the Variance of the Adaptive Learning Rate and Beyond , 2019, ICLR.

[2]  Yoshua Bengio,et al.  Generative Adversarial Nets , 2014, NIPS.

[3]  K. C. Huang,et al.  Cell size and growth regulation in the Arabidopsis thaliana apical stem cell niche , 2016, Proceedings of the National Academy of Sciences.

[4]  Ullrich Köthe,et al.  Ilastik: Interactive learning and segmentation toolkit , 2011, 2011 IEEE International Symposium on Biomedical Imaging: From Nano to Macro.

[5]  Marius Pachitariu,et al.  Cellpose: a generalist algorithm for cellular segmentation , 2020, Nature methods.

[6]  Tero Karras,et al.  Training Generative Adversarial Networks with Limited Data , 2020, NeurIPS.

[7]  E. Meijering A bird’s-eye view of deep learning in bioimage analysis , 2020, Computational and structural biotechnology journal.

[8]  Wolfgang Alt,et al.  Generalized Voronoi Tessellation as a Model of Two-dimensional Cell Tissue Dynamics , 2009, Bulletin of mathematical biology.

[9]  Eugene W. Myers,et al.  Biobeam—Multiplexed wave-optical simulations of light-sheet microscopy , 2017, PLoS Comput. Biol..

[10]  Johannes Stegmaier,et al.  Spherical Harmonics For Shape-Constrained 3d Cell Segmentation , 2020, 2021 IEEE 18th International Symposium on Biomedical Imaging (ISBI).

[11]  Johannes Stegmaier,et al.  SEGMENT3D: A web-based application for collaborative segmentation of 3D images used in the shoot apical meristem , 2017, 2018 IEEE 15th International Symposium on Biomedical Imaging (ISBI 2018).

[12]  P. Koumoutsakos,et al.  MorphoGraphX: A platform for quantifying morphogenesis in 4D , 2015, eLife.

[13]  Johannes Stegmaier,et al.  Towards Annotation-Free Segmentation of Fluorescently Labeled Cell Membranes in Confocal Microscopy Images , 2019, SASHIMI@MICCAI.

[14]  Johannes Stegmaier,et al.  CNN-Based Preprocessing to Optimize Watershed-Based Cell Segmentation in 3D Confocal Microscopy Images , 2018, 2019 IEEE 16th International Symposium on Biomedical Imaging (ISBI 2019).

[15]  Ralf Mikut,et al.  Semi-Automatic Generation Of Tight Binary Masks And Non-Convex Isosurfaces For Quantitative Analysis Of 3d Biological Samples , 2020, 2020 IEEE International Conference on Image Processing (ICIP).

[16]  Lassi Paavolainen,et al.  nucleAIzer: A Parameter-free Deep Learning Framework for Nucleus Segmentation Using Image Style Transfer , 2020, Cell systems.

[17]  Hans-Peter Meinzer,et al.  Statistical shape models for 3D medical image segmentation: A review , 2009, Medical Image Anal..

[18]  Simon Osindero,et al.  Conditional Generative Adversarial Nets , 2014, ArXiv.

[19]  Karl Rohr,et al.  A spherical harmonics intensity model for 3D segmentation and 3D shape analysis of heterochromatin foci , 2016, Medical Image Anal..

[20]  Daniel Rueckert,et al.  Real-Time Single Image and Video Super-Resolution Using an Efficient Sub-Pixel Convolutional Neural Network , 2016, 2016 IEEE Conference on Computer Vision and Pattern Recognition (CVPR).

[21]  Karel Svoboda,et al.  A platform for brain-wide imaging and reconstruction of individual neurons , 2016, eLife.

[22]  David Svoboda,et al.  MitoGen: A Framework for Generating 3D Synthetic Time-Lapse Sequences of Cell Populations in Fluorescence Microscopy , 2017, IEEE Transactions on Medical Imaging.

[23]  Jens C. Otte,et al.  Fast Segmentation of Stained Nuclei in Terabyte-Scale, Time Resolved 3D Microscopy Image Stacks , 2014, PloS one.

[24]  David Svoboda,et al.  On Generative Modeling of Cell Shape Using 3D GANs , 2019, ICIAP.

[25]  Johannes Stegmaier,et al.  Robust 3D Cell Segmentation: Extending the View of Cellpose , 2021, ArXiv.

[26]  Anne E Carpenter,et al.  CellProfiler 3.0: Next-generation image processing for biology , 2018, PLoS biology.

[27]  Mike Heilemann,et al.  SuReSim: simulating localization microscopy experiments from ground truth models , 2016, Nature Methods.

[28]  Jean-Christophe Olivo-Marin,et al.  Characterization of cell shape and deformation in 3D using Spherical Harmonics , 2012, 2012 9th IEEE International Symposium on Biomedical Imaging (ISBI).

[29]  Nathalie Harder,et al.  An Objective Comparison of Cell Tracking Algorithms , 2017, Nature Methods.

[30]  Nils Norlin,et al.  Inverted light-sheet microscope for imaging mouse pre-implantation development , 2015, Nature Methods.

[31]  E. Delp,et al.  Three Dimensional Synthetic Non-Ellipsoidal Nuclei Volume Generation Using Bézier Curves , 2021, 2021 IEEE 18th International Symposium on Biomedical Imaging (ISBI).

[32]  Alexei A. Efros,et al.  Image-to-Image Translation with Conditional Adversarial Networks , 2016, 2017 IEEE Conference on Computer Vision and Pattern Recognition (CVPR).

[33]  Randy Heiland,et al.  PhysiCell: An open source physics-based cell simulator for 3-D multicellular systems , 2017, bioRxiv.

[34]  P. Bourgine,et al.  A workflow to process 3D+time microscopy images of developing organisms and reconstruct their cell lineage , 2016, Nature Communications.

[35]  Thomas Brox,et al.  3D U-Net: Learning Dense Volumetric Segmentation from Sparse Annotation , 2016, MICCAI.

[36]  Erik Meijering,et al.  Imagining the future of bioimage analysis , 2016, Nature Biotechnology.