Deep-learning-based label-free segmentation of cell nuclei in time-lapse refractive index tomograms

In order to identify cell nuclei, fluorescent proteins or staining agents has been widely used. However, use of exogenous agents inevitably prevents from long-term imaging of live cells and rapid analysis, and even interferes with intrinsic physiological conditions. In this work, we proposed a method of label-free segmentation of cell nuclei in optical diffraction tomography images by exploiting a deep learning framework. The proposed method was applied for precise cell nucleus segmentation in two, three, and four-dimensional label-free imaging. A novel architecture with optimised training strategies was validated through cross-modality and cross-laboratory experiments. The proposed method would bring out broad and immediate biomedical applications with our framework publicly available.

[1]  Zhuowen Tu,et al.  Aggregated Residual Transformations for Deep Neural Networks , 2016, 2017 IEEE Conference on Computer Vision and Pattern Recognition (CVPR).

[2]  Michael I. Jordan,et al.  Advances in Neural Information Processing Systems 30 , 1995 .

[3]  C. Depeursinge,et al.  Quantitative phase imaging in biomedicine , 2012, 2012 Conference on Lasers and Electro-Optics (CLEO).

[4]  Nicolas Pavillon,et al.  Noninvasive detection of macrophage activation with single-cell resolution through machine learning , 2018, Proceedings of the National Academy of Sciences.

[5]  Kyoohyun Kim,et al.  Optical diffraction tomography techniques for the study of cell pathophysiology , 2016, 1603.00592.

[6]  Sung-Hee Hong,et al.  Characterizations of individual mouse red blood cells parasitized by Babesia microti using 3-D holographic microscopy , 2015, Scientific Reports.

[7]  Steven Frank Kemeny,et al.  A Simplified Implementation of Edge Detection in MATLAB is Faster and More Sensitive than Fast Fourier Transform for Actin Fiber Alignment Quantification , 2011, Microscopy and Microanalysis.

[8]  YongKeun Park,et al.  Label-free non-invasive quantitative measurement of lipid contents in individual microalgal cells using refractive index tomography , 2017, Scientific Reports.

[9]  Jonghee Yoon,et al.  Holographic deep learning for rapid optical screening of anthrax spores , 2017, Science Advances.

[10]  YongKeun Park,et al.  Identification of non-activated lymphocytes using three-dimensional refractive index tomography and machine learning , 2017, Scientific Reports.

[11]  Sergey Ioffe,et al.  Batch Normalization: Accelerating Deep Network Training by Reducing Internal Covariate Shift , 2015, ICML.

[12]  Yuan Zhang,et al.  Retinal vessel segmentation of color fundus images using multiscale convolutional neural network with an improved cross-entropy loss function , 2018, Neurocomputing.

[13]  Gabriel Popescu,et al.  Quantitative Phase Imaging , 2012 .

[14]  YongKeun Park,et al.  Holotomography: refractive index as an intrinsic imaging contrast for 3-D label-free live cell imaging , 2017, bioRxiv.

[15]  Jonghee Yoon,et al.  Optical diffraction tomography using a digital micromirror device for stable measurements of 4D refractive index tomography of cells , 2016, SPIE BiOS.

[16]  Thomas Brox,et al.  U-Net: Convolutional Networks for Biomedical Image Segmentation , 2015, MICCAI.

[17]  Tara N. Sainath,et al.  Improving deep neural networks for LVCSR using rectified linear units and dropout , 2013, 2013 IEEE International Conference on Acoustics, Speech and Signal Processing.

[18]  Hyun-seok Min,et al.  Quantitative Phase Imaging and Artificial Intelligence: A Review , 2018, IEEE Journal of Selected Topics in Quantum Electronics.

[19]  J Kapuscinski,et al.  DAPI: a DNA-specific fluorescent probe. , 1995, Biotechnic & histochemistry : official publication of the Biological Stain Commission.

[20]  YoungJu Jo,et al.  Quantitative Phase Imaging Techniques for the Study of Cell Pathophysiology: From Principles to Applications , 2013, Sensors.

[21]  Won-Ki Jeong,et al.  FusionNet: A Deep Fully Residual Convolutional Neural Network for Image Segmentation in Connectomics , 2016, Frontiers in Computer Science.

[22]  YongKeun Park,et al.  Profiling individual human red blood cells using common-path diffraction optical tomography , 2014, Scientific Reports.

[23]  YongKeun Park,et al.  Learning-based screening of hematologic disorders using quantitative phase imaging of individual red blood cells. , 2019, Biosensors & bioelectronics.

[24]  E. Wolf Three-dimensional structure determination of semi-transparent objects from holographic data , 1969 .

[25]  Geoffrey E. Hinton,et al.  ImageNet classification with deep convolutional neural networks , 2012, Commun. ACM.

[26]  Yen-Ping Chu,et al.  Edge Enhancement Nucleus and Cytoplast Contour Detector of Cervical Smear Images , 2008, IEEE Transactions on Systems, Man, and Cybernetics, Part B (Cybernetics).

[27]  Minh N. Do,et al.  Automatic Gleason grading of prostate cancer using quantitative phase imaging and machine learning , 2017, Journal of biomedical optics.

[28]  S. Shapshay,et al.  Detection of preinvasive cancer cells , 2000, Nature.

[29]  Michael B. Wallace,et al.  Observation of periodic fine structure in reflectance from biological tissue: A new technique for measuring nuclear size distribution , 1998 .

[30]  Kyoohyun Kim,et al.  Intracellular mass density increase is accompanying but not sufficient for stiffening and growth arrest of yeast cells , 2018, bioRxiv.

[31]  Rob Fergus,et al.  Visualizing and Understanding Convolutional Networks , 2013, ECCV.

[32]  Ata Mahjoubfar,et al.  Deep Learning in Label-free Cell Classification , 2016, Scientific Reports.

[34]  Jimmy Ba,et al.  Adam: A Method for Stochastic Optimization , 2014, ICLR.

[35]  Xiangyu Zhang,et al.  Large Kernel Matters — Improve Semantic Segmentation by Global Convolutional Network , 2017, 2017 IEEE Conference on Computer Vision and Pattern Recognition (CVPR).

[36]  R. Howe,et al.  17th International Conference on Medical Image Computing and Computer-Assisted Intervention. , 2014, Medical image computing and computer-assisted intervention : MICCAI ... International Conference on Medical Image Computing and Computer-Assisted Intervention.

[37]  Kyoohyun Kim,et al.  Three-dimensional label-free imaging and quantification of lipid droplets in live hepatocytes , 2016, Scientific Reports.

[38]  Martin A. Riedmiller,et al.  A direct adaptive method for faster backpropagation learning: the RPROP algorithm , 1993, IEEE International Conference on Neural Networks.

[39]  G. Ahn,et al.  Computational Modeling and Clonogenic Assay for Radioenhancement of Gold Nanoparticles Using 3D Live Cell Images , 2018, Radiation Research.