Segmentation of fluorescence microscopy images using three dimensional active contours with inhomogeneity correction

Image segmentation is an important step in the quantitative analysis of fluorescence microscopy data. Since fluorescence microscopy volumes suffer from intensity inhomogeneity, low image contrast and limited depth resolution, poor edge details, and irregular structure shape, segmentation still remains a challenging problem. This paper describes a nuclei segmentation method for fluorescence microscopy based on the use of three dimensional (3D) active contours with inhomogeneity correction. The correction information utilizes 3D volume information while addressing intensity inhomogeneity across vertical and horizontal directions. Experimental results demonstrate that the proposed method achieves better performance than other reported methods

Tubule Segmentation of Fluorescence Microscopy Images Based on Convolutional Neural Networks With Inhomogeneity Correction

Fluorescence microscopy has become a widely used tool for studying various biological structures of in vivo tissue or cells. However, quantitative analysis of these biological structures remains a challenge due to their complexity which is exacerbated by distortions caused by lens aberrations and light scattering. Moreover, manual quantification of such image volumes is an intractable and error-prone process, making the need for automated image analysis methods crucial. This paper describes a segmentation method for tubular structures in fluorescence microscopy images using convolutional neural networks with data augmentation and inhomogeneity correction. The segmentation results of the proposed method are visually and numerically compared with other microscopy segmentation methods. Experimental results indicate that the proposed method has better performance with correctly segmenting and identifying multiple tubular structures compared to other methods

DeepSynth: Three-dimensional nuclear segmentation of biological images using neural networks trained with synthetic data

The scale of biological microscopy has increased dramatically over the past ten years, with the development of new modalities supporting collection of high-resolution fluorescence image volumes spanning hundreds of microns if not millimeters. The size and complexity of these volumes is such that quantitative analysis requires automated methods of image processing to identify and characterize individual cells. For many workflows, this process starts with segmentation of nuclei that, due to their ubiquity, ease-of-labeling and relatively simple structure, make them appealing targets for automated detection of individual cells. However, in the context of large, three-dimensional image volumes, nuclei present many challenges to automated segmentation, such that conventional approaches are seldom effective and/or robust. Techniques based upon deep-learning have shown great promise, but enthusiasm for applying these techniques is tempered by the need to generate training data, an arduous task, particularly in three dimensions. Here we present results of a new technique of nuclear segmentation using neural networks trained on synthetic data. Comparisons with results obtained using commonly-used image processing packages demonstrate that DeepSynth provides the superior results associated with deep-learning techniques without the need for manual annotation

Three Dimensional Fluorescence Microscopy Image Synthesis and Segmentation

Advances in fluorescence microscopy enable acquisition of 3D image volumes with better image quality and deeper penetration into tissue. Segmentation is a required step to characterize and analyze biological structures in the images and recent 3D segmentation using deep learning has achieved promising results. One issue is that deep learning techniques require a large set of groundtruth data which is impractical to annotate manually for large 3D microscopy volumes. This paper describes a 3D deep learning nuclei segmentation method using synthetic 3D volumes for training. A set of synthetic volumes and the corresponding groundtruth are generated using spatially constrained cycle-consistent adversarial networks. Segmentation results demonstrate that our proposed method is capable of segmenting nuclei successfully for various data sets

Three Dimensional Blind Image Deconvolution for Fluorescence Microscopy using Generative Adversarial Networks

Due to image blurring image deconvolution is often used for studying biological structures in fluorescence microscopy. Fluorescence microscopy image volumes inherently suffer from intensity inhomogeneity, blur, and are corrupted by various types of noise which exacerbate image quality at deeper tissue depth. Therefore, quantitative analysis of fluorescence microscopy in deeper tissue still remains a challenge. This paper presents a three dimensional blind image deconvolution method for fluorescence microscopy using 3way spatially constrained cycle-consistent adversarial networks. The restored volumes of the proposed deconvolution method and other well-known deconvolution methods, denoising methods, and an inhomogeneity correction method are visually and numerically evaluated. Experimental results indicate that the proposed method can restore and improve the quality of blurred and noisy deep depth microscopy image visually and quantitatively

Three Dimensional Nuclei Segmentation and Classification of Fluorescence Microscopy Images

Segmentation and classification of cell nuclei in fluorescence 3D microscopy image volumes are fundamental steps for image analysis. However, accurate cell nuclei segmentation and detection in microscopy image volumes are hampered by poor image quality, crowding of nuclei, and large variation in nuclei size and shape. In this paper, we present an unsupervised volume to volume translation approach adapted from the Recycle-GAN using modified Hausdorff distance loss for synthetically generating nuclei with better shapes. A 3D CNN with a regularization term is used for nuclei segmentation and classification followed by nuclei boundary refinement. Experimental results demonstrate that the proposed method can successfully segment nuclei and identify individual nuclei

Segmentation and Deconvolution of Fluorescence Microscopy Volumes

Recent advances in optical microscopy have enabled biologists collect fluorescence microscopy volumes cellular and subcellular structures of living tissue. This results in collecting large datasets of microscopy volume and needs image processing aided automated quantification method. To quantify biological structures a first and fundamental step is segmentation. Yet, the quantitative analysis of the microscopy volume is hampered by light diffraction, distortion created by lens aberrations in different directions, complex variation of biological structures. This thesis describes several proposed segmentation methods to identify various biological structures such as nuclei or tubules observed in fluorescence microscopy volumes. To achieve nuclei segmentation, multiscale edge detection method and 3D active contours with inhomogeneity correction method are used for segmenting nuclei. Our proposed 3D active contours with inhomogeneity correction method utilizes 3D microscopy volume information while addressing intensity inhomogeneity across vertical and horizontal directions. To achieve tubules segmentation, ellipse model fitting to tubule boundary method and convolutional neural networks with inhomogeneity correction method are performed. More specifically, ellipse fitting method utilizes a combination of adaptive and global thresholding, potentials, z direction refinement, branch pruning, end point matching, and boundary fitting steps to delineate tubular objects. Also, the deep learning based method combines intensity inhomogeneity correction, data augmentation, followed by convolutional neural networks architecture. Moreover, this thesis demonstrates a new deconvolution method to improve microscopy image quality without knowing the 3D point spread function using a spatially constrained cycle-consistent adversarial networks. The results of proposed methods are visually and numerically compared with other methods. Experimental results demonstrate that our proposed methods achieve better performance than other methods for nuclei/tubules segmentation as well as deconvolution

Hard X-ray free-electron laser with femtosecond-scale timing jitter

The hard X-ray free-electron laser at the Pohang Accelerator Laboratory (PAL-XFEL) in the Republic of Korea achieved saturation of a 0.144 nm free-electron laser beam on 27 November 2016, making it the third hard X-ray free-electron laser in the world, following the demonstrations of the Linac Coherent Light Source (LCLS) and the SPring-8 Angstrom Compact Free Electron Laser (SACLA). The use of electron-beam-based alignment incorporating undulator radiation spectrum analysis has allowed reliable operation of PAL-XFEL with unprecedented temporal stability and dispersion-free orbits. In particular, a timing jitter of just 20 fs for the free-electron laser photon beam is consistently achieved due to the use of a state-of-the-art design of the electron linear accelerator and electron-beam-based alignment. The low timing jitter of the electron beam makes it possible to observe Bi(111) phonon dynamics without the need for timing-jitter correction, indicating that PAL-XFEL will be an extremely useful tool for hard X-ray time-resolved experiments.1143Nsciescopu