Functional optical imaging in neuroscience is rapidly growing with the development of new optical systems and fluorescence indicators. To realize the potential of these massive spatiotemporal datasets for relating neuronal activity to behavior and stimuli and uncovering local circuits in the brain, accurate automated processing is increasingly essential. In this review, we cover recent computational developments in the full data processing pipeline of functional optical microscopy for neuroscience data and discuss ongoing and emerging challenges.
Providing high-quality electron images and hyperspectral X-ray maps is a focus of many modern electron microscopy laboratories. Nevertheless, further image processing and annotations are often needed to prepare them for publications and reports. For multi-user facilities, accessibility to processing software can be a limitation either through license costs or availability of processing stations. Open-source software running on multiple platforms allows for post-acquisition data processing in-lab or on user-owned devices. We developed Probelab ReImager to supersede our vendor-supplied acquisiti…
University of Chicago（芝加哥大学）；Argonne National Lab（阿贡国家实验室）
Technological advancements in modern scientific instruments, such as scanning electron microscopes (SEMs), have significantly increased data acquisition rates and image resolutions enabling new questions to be explored; however, the resulting data volumes and velocities, combined with automated experiments, are quickly overwhelming scientists as there remain crucial steps that require human intervention, for example reviewing image focus. We present a fast out-of-focus detection algorithm for electron microscopy images collected serially and demonstrate that it can be used to provide near-real…
University of Cambridge（剑桥大学）；Imperial College London（伦敦帝国理工学院）
Understanding how networks of neurons process information is one of the key challenges in modern neuroscience. A necessary step to achieve this goal is to be able to observe the dynamics of large populations of neurons over a large area of the brain. Light-field microscopy (LFM), a type of scanless microscope, is a particularly attractive candidate for high-speed three-dimensional (3D) imaging. It captures volumetric information in a single snapshot, allowing volumetric imaging at video frame-rates. Specific features of imaging neuronal activity using LFM call for the development of novel mach…
We present a fully modular and scalable software pipeline for processing electron microscope (EM) images of brain slices into 3D visualization of individual neurons and demonstrate an end-to-end segmentation of a large EM volume using a supercomputer. Our pipeline scales multiple packages used by the EM community with minimal changes to the original source codes. We tested each step of the pipeline individually, on a workstation, a cluster, and a supercomputer. Furthermore, we can compose workflows from these operations using a Balsam database that can be triggered during the data acquisition …
This book chapter reviews progress in crystallographic image processing (CIP) for scanning probe microscopy (SPM) that has occurred since our description of the technique was first put into open access in this book series in the year 2010. The signal to noise ratio in all kinds of experimental images of more or less regular 2D periodic arrays is significantly enhanced by CIP and the technique is independent of the type of recording device. In the SPM imaging context, CIP can be understood as an a posteriori sharpening of the effective experimental scanning probe tip by computational means. It …
Université de Tunis El Manar（突尼斯埃尔马纳尔大学）；Faculté des Sciences de Tunis（突尼斯科学学院）
Cells segmentation shows rapid growth in biology. Indeed, using the classical segmentation methods only is not enough to segment this type of images. In this manuscript, we will present a new method of ribosomes segmentation. A pre-treatment phase will precedes the segmentation process and after that a post-processing will proceed.
Today the gold standard for in vivo imaging through scattering tissue is the point-scanning two-photon microscope (PSTPM). Especially in neuroscience, PSTPM is widely used for deep-tissue imaging in the brain. However, due to sequential scanning, PSTPM is slow. Temporal focusing microscopy (TFM), on the other hand, focuses femtosecond pulsed laser light temporally, while keeping wide-field illumination, and is consequently much faster. However, due to the use of a camera detector, TFM suffers from the scattering of emission photons. As a result, TFM produces images of poor spatial resolution a…
Manual preparation of tissue slices for microscopy imaging can introduce tissue tears and overlaps. Typically, further digital processing algorithms such as registration and 3D reconstruction from tissue image stacks cannot handle images with tissue tear/overlap artifacts, and so such images are usually discarded. In this paper, we propose an imaging model and an algorithm to digitally separate overlapping tissue data of mouse brain images into two layers. We show the correctness of our model and the algorithm by comparing our results with the ground truth.
Lawrence Berkeley National Laboratory（劳伦斯伯克利国家实验室）；National Renewable Energy Laboratory（美国国家可再生能源实验室）；Northeastern University（东北大学）
An image dataset of 10 different size molecules, where each molecule has 2,000 structural variants, is generated from the 2D cross-sectional projection of Molecular Dynamics trajectories. The purpose of this dataset is to provide a benchmark dataset for the increasing need of machine learning, deep learning and image processing on the study of scattering, imaging and microscopy.
To obtain microscope images at multiple focal planes, the distance between the objective and sample can be mechanically adjusted. Images are acquired sequentially at each axial distance. Digital refocusing with a light-emitting diode (LED) array microscope allows elimination of this mechanical movement. In an LED array microscope, the light source of a conventional widefield microscope is replaced with a 2-dimensional LED matrix. A stack of images is acquired from the LED array microscope by sequentially illuminating each LED and capturing an image. Previous work has shown that we can achieve …
As an important branch of photoacoustic microscopy, optical-resolution photoacoustic microscopy suffers from limited depth of field due to the strongly focused laser beam. In this work, a 3D information fusion algorithm based on 3D stationary wavelet transform and joint weighted evaluation optimization is proposed to fuse multi-focus photoacoustic data to achieve large-volumetric and high-resolution 3D imaging. First, a three-dimensional stationary wavelet transform was performed on the multi-focus data to obtain eight wavelet coefficients. Differential evolution algorithm based on joint weigh…
University of California Berkeley（加州大学伯克利分校）；Lawrence Berkeley National Laboratory（劳伦斯伯克利国家实验室）
In the field of transmission electron microscopy, data interpretation often lags behind acquisition methods, as image processing methods often have to be manually tailored to individual datasets. Machine learning offers a promising approach for fast, accurate analysis of electron microscopy data. Here, we demonstrate a flexible two step pipeline for analysis of high resolution transmission electron microscopy data, which uses a U-Net for segmentation followed by a random forest for detection of stacking faults. Our trained U-Net is able to segment nanoparticle regions from amorphous background…
