3D vessel-wall virtual histology of whole-body perfused mice using a novel heavy element stain.
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ABSTRACT:
SUBMITTER: Dunmore-Buyze PJ
PROVIDER: S-EPMC6345940 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:Histological investigations are indispensable with regards to the identification of structural tissue details but are limited to two-dimensional images, which are often visualized in one and the same plane for comparison reasons. Nondestructive three-dimensional technologies such as X-ray micro- and nanoCT have proven to provide valuable benefits for the understanding of anatomical structures as they allow visualization of structural details in 3D and from arbitrary viewing angles. Nevertheless, low attenuation of soft tissue has hampered their application in the field of 3D virtual histology. We present a hematein-based X-ray staining method that specifically targets the cell nuclei of cells, as demonstrated for a whole liver lobule of a mouse. Combining the novel staining protocol with the high resolving power of a recently developed nanoCT system enables the 3D visualization of tissue architecture in the nanometer range, thereby revealing the real 3D morphology and spatial distribution of the cell nuclei. Furthermore, our technique is compatible with conventional histology, as microscopic slides can be derived from the very same stained soft-tissue sample and further counter staining is possible. Thus, our methodology demonstrates future applicability for modern histopathology using laboratory X-ray CT devices.
| S-EPMC6294809 | biostudies-other
Project description:PurposeTo develop a respiratory motion correction framework to accelerate free-breathing three-dimensional (3D) whole-heart coronary lumen and coronary vessel wall MRI.MethodsWe developed a 3D flow-independent approach for vessel wall imaging based on the subtraction of data with and without T2-preparation prepulses acquired interleaved with image navigators. The proposed method corrects both datasets to the same respiratory position using beat-to-beat translation and bin-to-bin nonrigid corrections, producing coregistered, motion-corrected coronary lumen and coronary vessel wall images. The proposed method was studied in 10 healthy subjects and was compared with beat-to-beat translational correction (TC) and no motion correction for the left and right coronary arteries. Additionally, the coronary lumen images were compared with a 6-mm diaphragmatic navigator gated and tracked scan.ResultsNo significant differences (P > 0.01) were found between the proposed method and the gated and tracked scan for coronary lumen, despite an average improvement in scan efficiency to 96% from 59%. Significant differences (P < 0.01) were found in right coronary artery vessel wall thickness, right coronary artery vessel wall sharpness, and vessel wall visual score between the proposed method and TC.ConclusionThe feasibility of a highly efficient motion correction framework for simultaneous whole-heart coronary lumen and vessel wall has been demonstrated. Magn Reson Med 77:1894-1908, 2017. © 2016 International Society for Magnetic Resonance in Medicine.
| S-EPMC5412916 | biostudies-literature
Project description:BackgroundThree-dimensional (3D) whole-brain vessel wall imaging (VWI) has demonstrated exquisite image quality for delineating intracranial atherosclerotic disease (ICAD) and reliability for quantifying normal vessel dimensions. However, its reliability in quantifying plaque morphology remains unknown.PurposeTo evaluate the plaque morphologic quantification reliability of 3D whole-brain VWI in patients via comparison with 3D targeted VWI and 2D turbo spin-echo (TSE).Study typeProspective.PopulationThirty-three patients with symptomatic ICAD.Field strength/sequenceA 3D and 2D TSE acquired at 3.0 T.AssessmentEach participant underwent two VWI sessions with an interval of 7-10 days. Three readers identified in consensus all the plaques on both whole-brain and targeted 3D VWI. Their lumen and vessel wall area and volume, plaque burden, percent stenosis, and vessel wall remodeling were measured for by two independent readers. At each culprit plaque determined by a radiologist, the lumen and vessel wall area, plaque burden, plaque-to-wall contrast ratio (CR), and plaque enhancement ratio (ER) were measured for 2D and 3D VWI methods.Statistical testsIntra-class correlation coefficient (ICC) was used to evaluate for 3D VWI's interobserver/intraobserver agreement, interscan repeatability, and agreement with 2D TSE in each plaque morphologic measurements. Paired t test was performed for detecting the differences in plaque-to-wall CR and plaque ER between the two 3D methods.ResultsEighty-four plaques were detected by both 3D VWI methods. Whole-brain VWI provided excellent interobserver/intraobserver agreement (ICCs: 0.79-0.99/0.95-0.99), interscan repeatability (ICCs: 0.85-0.99), agreement with 2D TSE (ICC: 0.80-0.94) in all morphologic measurements. ICCs of whole-brain VWI (0.79-0.99) were higher or equal to those of targeted VWI (0.76-0.99). The plaque-to-wall CR and plaque ER were significantly higher on whole-brain VWI than on targeted VWI.Data conclusionThe 3D whole-brain VWI provides excellent interobserver/intraobserver agreement, interscan repeatability, and agreement with 2D TSE in plaque morphologic quantification of ICAD and outperforms 3D targeted VWI.Level of evidence2 TECHNICAL EFFICACY STAGE: 2.
| S-EPMC8812367 | biostudies-literature
Project description:BACKGROUND:One of the potentially important applications of three-dimensional (3D) intracranial vessel wall (IVW) cardiovascular magnetic resonance (CMR) is to monitor disease progression and regression via quantitative measurement of IVW morphology during medical management or drug development. However, a prerequisite for this application is to validate that IVW morphologic measurements based on the modality are reliable. In this study we performed comprehensive reliability analysis for the recently proposed whole-brain IVW CMR technique. METHODS:Thirty-four healthy subjects and 10 patients with known intracranial atherosclerotic disease underwent repeat whole-brain IVW CMR scans. In 19 of the 34 subjects, two-dimensional (2D) turbo spin-echo (TSE) scan was performed to serve as a reference for the assessment of vessel dimensions. Lumen and wall volume, normalized wall index, mean and maximum wall thickness were measured in both 3D and 2D IVW CMR images. Scan-rescan, intra-observer, and inter-observer reproducibility of 3D IVW CMR in the quantification of IVW or plaque dimensions were respectively assessed in volunteers and patients as well as for different healthy subjectsub-groups (i.e. < 50 and ≥ 50 years). The agreement in vessel wall and lumen measurements between the 3D technique and the 2D TSE method was also investigated. In addition, the sample size required for future longitudinal clinical studies was calculated. RESULTS:The intra-class correlation coefficient (ICC) and Bland-Altman plots indicated excellent reproducibility and inter-method agreement for all morphologic measurements (All ICCs > 0.75). In addition, all ICCs of patients were equal to or higher than that of healthy subjects except maximum wall thickness. In volunteers, all ICCs of the age group of ≥50 years were equal to or higher than that of the age group of < 50 years. Normalized wall index and mean and maximum wall thickness were significantly larger in the age group of ≥50 years. To detect 5% - 20% difference between placebo and treatment groups, normalized wall index requires the smallest sample size while lumen volume requires the highest sample size. CONCLUSIONS:Whole-brain 3D IVW CMR is a reliable imaging method for the quantification of intracranial vessel dimensions and could potentially be useful for monitoring plaque progression and regression.
| S-EPMC6000985 | biostudies-literature
Project description:Many histological methods require staining of the cytoplasm, which provides instrumental details for diagnosis. One major limitation is the production of 2D images obtained by destructive preparation of 3D tissue samples. X-ray absorption micro- and nanocomputed tomography (microCT and nanoCT) allows for a nondestructive investigation of a 3D tissue sample, and thus aids to determine regions of interest for further histological examinations. However, application of microCT and nanoCT to biological samples (e.g., biopsies) is limited by the missing contrast within soft tissue, which is important to visualize morphological details. We describe an eosin-based preparation overcoming the challenges of contrast enhancement and selectivity for certain tissues. The eosin-based staining protocol is suitable for whole-organ staining, which then enables high-resolution microCT imaging of whole organs and nanoCT imaging of smaller tissue pieces retrieved from the original sample. Our results demonstrate suitability of the eosin-based staining method for diagnostic screening of 3D tissue samples without impeding further diagnostics through histological methods.
| S-EPMC5877988 | biostudies-literature
Project description:Bacterial endospores are some of the most resilient forms of life known to us, with their persistent survival capability resulting from a complex and effective structural organization. The outer membrane of endospores is surrounded by the densely packed endospore coat and exosporium, containing amyloid or amyloid-like proteins. In fact, it is the impenetrable composition of the endospore coat and the exosporium that makes staining methodologies for endospore detection complex and challenging. Therefore, a plausible strategy for facile and expedient staining would be to target components of the protective surface layers of the endospores. Instead of targeting endogenous markers encapsulated in the spores, here we demonstrated staining of these dormant life entities that targets the amyloid domains, i.e., the very surface components that make the coats of these species impenetrable. Using an amyloid staining dye, thioflavin T (ThT), we examined this strategy. A short incubation of bacillus endospore suspensions with ThT, under ambient conditions, resulted in (i) an enhancement of the fluorescence of ThT and (ii) the accumulation of ThT in the endospores, affording fluorescence images with excellent contrast ratios. Fluorescence images revealed that ThT tends to accumulate in the surface regions of the endospores. The observed fluorescence enhancement and dye accumulation, coupled with the sensitivity of emission techniques, provide an effective and rapid means of staining endospores without the inconvenience of pre- or posttreatment of samples.
| S-EPMC3147746 | biostudies-literature
Project description:High-spatial-resolution histology of coronary artery autopsy samples play an important role for understanding heart disease such as myocardial infarction. Unfortunately, classical histology is often destructive, has thick slicing, requires extensive sample preparation, and is time-consuming. X-ray micro-CT provides fast nondestructive 3D imaging but absorption contrast is often insufficient, especially for observing soft-tissue features with high resolution. Here we show that propagation-based x-ray phase-contrast tomography has the resolution and contrast to image clinically relevant soft-tissue features in intact coronary artery autopsy samples with cellular resolution. We observe microscopic lipid-rich plaques, individual adipose cells, ensembles of few foam cells, and the thin fibrous cap. The method relies on a small-spot laboratory x-ray microfocus source, and provides high-spatial resolution in all three dimensions, fast data acquisition, minimum sample distortion and requires no sample preparation.
| S-EPMC6054690 | biostudies-other
Project description:Purpose: Surgery is an essential part of the curative plan for most patients affected with solid tumors. The outcome of such surgery, e.g., recurrence rates and ultimately patient survival, depends on several factors where the resection margin is of key importance. Presently, the resection margin is assessed by classical histology, which is time-consuming (several days), destructive, and basically only gives two-dimensional information. Clearly, it would be advantageous if immediate feedback on tumor extension in all three dimensions were available to the surgeon intraoperatively. Approach: We investigate a laboratory propagation-based phase-contrast x-ray computed tomography system that provides the resolution, the contrast, and, potentially, the speed for this purpose. The system relies on a liquid-metal jet microfocus source and a scintillator-coated CMOS detector. Our study is performed on paraffin-embedded non-stained samples of human pancreatic neuroendocrine tumors, liver intrahepatic cholangiocarcinoma, and pancreatic serous cystic neoplasm (benign). Results: We observe tumors with distinct and sharp edges having cellular resolution ( ∼10 μm ) as well as many assisting histological landmarks, allowing for resection margin assessment. All x-ray data are compared with classical histology. The agreement is excellent. Conclusion: We conclude that the method has potential for intraoperative three-dimensional virtual histology.
| S-EPMC8820384 | biostudies-literature
Project description:BackgroundVirtual reality (VR) enables data visualization in an immersive and engaging manner, and it can be used for creating ways to explore scientific data. Here, we use VR for visualization of 3D histology data, creating a novel interface for digital pathology to aid cancer research.MethodsOur contribution includes 3D modeling of a whole organ and embedded objects of interest, fusing the models with associated quantitative features and full resolution serial section patches, and implementing the virtual reality application. Our VR application is multi-scale in nature, covering two object levels representing different ranges of detail, namely organ level and sub-organ level. In addition, the application includes several data layers, including the measured histology image layer and multiple representations of quantitative features computed from the histology.ResultsIn our interactive VR application, the user can set visualization properties, select different samples and features, and interact with various objects, which is not possible in the traditional 2D-image view used in digital pathology. In this work, we used whole mouse prostates (organ level) with prostate cancer tumors (sub-organ objects of interest) as example cases, and included quantitative histological features relevant for tumor biology in the VR model.ConclusionsOur application enables a novel way for exploration of high-resolution, multidimensional data for biomedical research purposes, and can also be used in teaching and researcher training. Due to automated processing of the histology data, our application can be easily adopted to visualize other organs and pathologies from various origins.
| S-EPMC8539837 | biostudies-literature
Project description:A multiscale three-dimensional (3D) virtual histology approach is presented, based on two configurations of propagation phase-contrast X-ray tomography, which have been implemented in close proximity at the GINIX endstation at the beamline P10/PETRA III (DESY, Hamburg, Germany). This enables the 3D reconstruction of characteristic morphological features of human pancreatic normal and tumor tissue, as obtained from cancer surgery, first in the form of a large-scale overview by parallel-beam illumination, followed by a zoom into a region-of-interest based on zoom tomography using a Kirkpatrick-Baez mirror with additional waveguide optics. To this end 1 mm punch biopsies of the tissue were taken. In the parallel tomography, a volumetric throughput on the order of 0.01 mm3 s-1 was achieved, while maintaining the ability to segment isolated cells. With a continuous rotation during the scan, a total acquisition time of less than 2 min was required for a full tomographic scan. Using the combination of both setups, islets of Langerhans, a three-dimensional cluster of cells in the endocrine part of the pancreas, could be located. Cells in such an islet were segmented and visualized in 3D. Further, morphological alterations of tumorous tissue of the pancreas were characterized. To this end, the anisotropy parameter Ω, based on intensity gradients, was used in order to quantify the presence of collagen fibers within the entire biopsy specimen. This proof-of-concept experiment of the multiscale approach on human pancreatic tissue paves the way for future 3D virtual pathology.
| S-EPMC7642968 | biostudies-literature