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A versatile pipeline for the multi-scale digital reconstruction and quantitative analysis of 3D tissue architecture.

ABSTRACT: A prerequisite for the systems biology analysis of tissues is an accurate digital three-dimensional reconstruction of tissue structure based on images of markers covering multiple scales. Here, we designed a flexible pipeline for the multi-scale reconstruction and quantitative morphological analysis of tissue architecture from microscopy images. Our pipeline includes newly developed algorithms that address specific challenges of thick dense tissue reconstruction. Our implementation allows for a flexible workflow, scalable to high-throughput analysis and applicable to various mammalian tissues. We applied it to the analysis of liver tissue and extracted quantitative parameters of sinusoids, bile canaliculi and cell shapes, recognizing different liver cell types with high accuracy. Using our platform, we uncovered an unexpected zonation pattern of hepatocytes with different size, nuclei and DNA content, thus revealing new features of liver tissue organization. The pipeline also proved effective to analyse lung and kidney tissue, demonstrating its generality and robustness.

SUBMITTER: Morales-Navarrete H

PROVIDER: S-EPMC4764584 | biostudies-literature | 2015 Dec

REPOSITORIES: biostudies-literature

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A versatile pipeline for the multi-scale digital reconstruction and quantitative analysis of 3D tissue architecture.

Morales-Navarrete Hernán H Segovia-Miranda Fabián F Klukowski Piotr P Meyer Kirstin K Nonaka Hidenori H Marsico Giovanni G Chernykh Mikhail M Kalaidzidis Alexander A Zerial Marino M Kalaidzidis Yannis Y

eLife 20151227

A prerequisite for the systems biology analysis of tissues is an accurate digital three-dimensional reconstruction of tissue structure based on images of markers covering multiple scales. Here, we designed a flexible pipeline for the multi-scale reconstruction and quantitative morphological analysis of tissue architecture from microscopy images. Our pipeline includes newly developed algorithms that address specific challenges of thick dense tissue reconstruction. Our implementation allows for a ...[more]

PMID: 26673893

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Project description:The three-dimensional (3D) configuration of chromosomes within the eukaryote nucleus is an important factor for several cellular functions, including gene expression regulation, and has also been linked with cancer-causing translocation events. While visualization of such architecture remains limited to low resolutions, the ability to infer structures at increasing resolutions has been enabled by recently-devised chromosome conformation capture techniques. In particular, when coupled with next generation sequencing, such methods yield an inventory of genome-wide chromatin contacts or interactions. Various algorithms have been advanced to operate on such contact data to produce reconstructed 3D configurations. Studies have shown that these reconstructions can provide added value over raw interaction data with respect to downstream biological insights. However, only limited, low-resolution reconstructions have been realized for mammals due to computational bottlenecks.Here we propose a two-stage algorithm to partially overcome these computational barriers. The central idea is to initially utilize existing reconstruction techniques on an individual chromosome basis, using intra-chromosomal contacts, and then to relatively position these chromosome-level reconstructions using inter-chromosomal contacts. This two-stage strategy represents a natural approach in view of the within- versus between- chromosome distribution of contacts. It can increase resolution ? 20 fold for mouse and human. After describing the algorithm we present 3D architectures for mouse embryonic stem cells and human lymphoblastoid cells. We evaluate the impact of several factors on reconstruction reproducibility and explore a variety of sampling schemes. We further analyze replicate data at differing resolutions obtained from recently devised in situ Hi-C assays. In all instances we demonstrate insensitivity of the whole-genome 3D reconstruction obtained by the two-stage algorithm to the sampling strategy used.Our two-stage algorithm has the potential to significantly increase the resolution of 3D genome reconstructions. The improvements are such that we can progress from 1 Mb resolution to 100 kb resolution, notable since this latter value has been identified as critical to inferring topological domains in analyses performed on the contact (rather than 3D) level.

Project description:PurposeFree-breathing Magnetization Transfer Contrast Bright blOOd phase SensiTive (MTC-BOOST) is a prototype balanced-Steady-State Free Precession sequence for 3D whole-heart imaging, that employs the endogenous magnetisation transfer contrast mechanism. This achieves reduction of flow and off-resonance artefacts, that often arise with the clinical T2prepared balanced-Steady-State Free Precession sequence, enabling high quality, contrast-agent free imaging of the thoracic cardiovascular anatomy. Fully-sampled MTC-BOOST acquisition requires long scan times (~10-24 min) and therefore acceleration is needed to permit its clinical incorporation. The aim of this study is to enable and clinically validate the 5-fold accelerated MTC-BOOST acquisition with joint Multi-Scale Variational Neural Network (jMS-VNN) reconstruction.MethodsThirty-six patients underwent free-breathing, 3D whole-heart imaging with the MTC-BOOST sequence, which is combined with variable density spiral-like Cartesian sampling and 2D image navigators for translational motion estimation. This sequence acquires two differently weighted bright-blood volumes in an interleaved fashion, which are then joined in a phase sensitive inversion recovery reconstruction to obtain a complementary fully co-registered black-blood volume. Data from eighteen patients were used for training, whereas data from the remaining eighteen patients were used for testing/evaluation. The proposed deep-learning based approach adopts a supervised multi-scale variational neural network for joint reconstruction of the two differently weighted bright-blood volumes acquired with the 5-fold accelerated MTC-BOOST. The two contrast images are stacked as different channels in the network to exploit the shared information. The proposed approach is compared to the fully-sampled MTC-BOOST and 5-fold undersampled MTC-BOOST acquisition with Compressed Sensing (CS) reconstruction in terms of scan/reconstruction time and bright-blood image quality. Comparison against conventional 2-fold undersampled T2-prepared 3D bright-blood whole-heart clinical sequence (T2prep-3DWH) is also included.ResultsAcquisition time was 3.0 ± 1.0 min for the 5-fold accelerated MTC-BOOST versus 9.0 ± 1.1 min for the fully-sampled MTC-BOOST and 11.1 ± 2.6 min for the T2prep-3DWH (p < 0.001 and p < 0.001, respectively). Reconstruction time was significantly lower with the jMS-VNN method compared to CS (10 ± 0.5 min vs 20 ± 2 s, p < 0.001). Image quality was higher for the proposed 5-fold undersampled jMS-VNN versus conventional CS, comparable or higher to the corresponding T2prep-3DWH dataset and similar to the fully-sampled MTC-BOOST.ConclusionThe proposed 5-fold accelerated jMS-VNN MTC-BOOST framework provides efficient 3D whole-heart bright-blood imaging in fast acquisition and reconstruction time with concomitant reduction of flow and off-resonance artefacts, that are frequently encountered with the clinical sequence. Image quality of the cardiac anatomy and thoracic vasculature is comparable or superior to the clinical scan and 5-fold CS reconstruction in faster reconstruction time, promising potential clinical adoption.

Dataset Information

A versatile pipeline for the multi-scale digital reconstruction and quantitative analysis of 3D tissue architecture.

Publications

A versatile pipeline for the multi-scale digital reconstruction and quantitative analysis of 3D tissue architecture.

Similar Datasets

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