Single-cell transcriptional dynamics in a living vertebrate.

The ability to quantify transcriptional dynamics in individual cells via live imaging has revolutionized our understanding of gene regulation. However, such measurements are lacking in the context of vertebrate embryos. We addressed this deficit by applying MS2-MCP mRNA labeling to the quantification of transcription in zebrafish, a model vertebrate. We developed a platform of transgenic organisms, light sheet fluorescence microscopy, and optimized image analysis that enables visualization and quantification of MS2 reporters. We used these tools to obtain the first single-cell, real-time measurements of transcriptional dynamics of the segmentation clock. Our measurements challenge the traditional view of smooth clock oscillations and instead suggest a model of discrete transcriptional bursts that are organized in space and time. Together, these results highlight how measuring single-cell transcriptional activity can reveal unexpected features of gene regulation and how this data can fuel the dialogue between theory and experiment.

OME-Zarr: a cloud-optimized bioimaging file format with international community support.

A growing community is constructing a next-generation file format (NGFF) for bioimaging to overcome problems of scalability and heterogeneity. Organized by the Open Microscopy Environment (OME), individuals and institutes across diverse modalities facing these problems have designed a format specification process (OME-NGFF) to address these needs. This paper brings together a wide range of those community members to describe the cloud-optimized format itself-OME-Zarr-along with tools and data resources available today to increase FAIR access and remove barriers in the scientific process. The current momentum offers an opportunity to unify a key component of the bioimaging domain-the file format that underlies so many personal, institutional, and global data management and analysis tasks.

OME-Zarr: a cloud-optimized bioimaging file format with international community support.

A growing community is constructing a next-generation file format (NGFF) for bioimaging to overcome problems of scalability and heterogeneity. Organized by the Open Microscopy Environment (OME), individuals and institutes across diverse modalities facing these problems have designed a format specification process (OME-NGFF) to address these needs. This paper brings together a wide range of those community members to describe the cloud-optimized format itself -- OME-Zarr -- along with tools and data resources available today to increase FAIR access and remove barriers in the scientific process. The current momentum offers an opportunity to unify a key component of the bioimaging domain -- the file format that underlies so many personal, institutional, and global data management and analysis tasks.

DaXi-high-resolution, large imaging volume and multi-view single-objective light-sheet microscopy.

The promise of single-objective light-sheet microscopy is to combine the convenience of standard single-objective microscopes with the speed, coverage, resolution and gentleness of light-sheet microscopes. We present DaXi, a single-objective light-sheet microscope design based on oblique plane illumination that achieves: (1) a wider field of view and high-resolution imaging via a custom remote focusing objective; (2) fast volumetric imaging over larger volumes without compromising image quality or necessitating tiled acquisition; (3) fuller image coverage for large samples via multi-view imaging and (4) higher throughput multi-well imaging via remote coverslip placement. Our instrument achieves a resolution of 450 nm laterally and 2 µm axially over an imaging volume of 3,000 × 800 × 300 µm. We demonstrate the speed, field of view, resolution and versatility of our instrument by imaging various systems, including Drosophila egg chamber development, zebrafish whole-brain activity and zebrafish embryonic development - up to nine embryos at a time.

An adaptive probabilistic atlas for anomalous brain segmentation in MR images.

PURPOSE: Automated segmentation of brain structures (objects) in MR three-dimensional (3D) images for quantitative analysis has been a challenge and probabilistic atlases (PAs) are among the most well-succeeded approaches. However, the existing models do not adapt to possible object anomalies due to the presence of a disease or a surgical procedure. Post-processing operation does not solve the problem, for example, tissue classification to detect and remove such anomalies inside the resulting segmentation mask, because segmentation errors on healthy tissues cannot be fixed. Such anomalies very often alter the shape and texture of the brain structures, making them different from the appearance of the model. In this paper, we present an effective and efficient adaptive probabilistic atlas, named AdaPro, to circumvent the problem and evaluate it on a challenging task - the segmentation of the left hemisphere, right hemisphere, and cerebellum, without pons and medulla, in 3D MR-T1 brain images of Epilepsy patients. This task is challenging due to temporal lobe resections, artifacts, and the absence of contrast in some parts between the structures of interest. METHODS: In AdaPro, we first build one probabilistic atlas per object of interest from a training set with normal 3D images and the corresponding 3D object masks. Second, we incorporate a texture classifier based on convex optimization which dynamically indicates the regions of the target 3D image where the PAs (shape constraints) should be further adapted. This strategy is mathematically more elegant and avoids problems with post-processing. Third, we add a new object-based delineation algorithm based on combinatorial optimization and diffusion filtering. AdaPro can then be used to locate and delineate the objects in the coordinate space of the atlas or of the test image. We also compare AdaPro with three other state-of-the-art methods: an statistical shape model based on synergistic object search and delineation, and two methods based on multi-atlas label fusion (MALF). RESULTS: We evaluate the methods quantitatively on 3D MR-T1 brain images of 2T and 3T from epilepsy patients, before and after temporal lobe resections, and on the template and native coordinate spaces. The results show that AdaPro is considerably faster and consistently more accurate than the baselines with statistical significance in both coordinate spaces. CONCLUSION: AdaPro can be used as a fast and effective step for brain tissue segmentation and it can also be easily extended to segment subcortical brain structures. By choice of its components, probabilistic atlas, texture classifier, and delineation algorithm, it can also be extended to other organs and imaging modalities.