Stereological quantification of microvessels using semiautomated evaluation of X-ray microtomography of hepatic vascular corrosion casts

Jiřík, Miroslav; Tonar, Zbyněk; Králíčková, Anna; Eberlová, Lada; Mírka, Hynek; Kochová, Petra; Gregor, Tomáš; Hošek, Petr; Svobodová, Miroslava; Rohan, Eduard; Králíčková, Milena; Liška, Václav

doi:10.1007/s11548-016-1378-3

Cited by 12 publications

(6 citation statements)

References 40 publications

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“…With recent advances in high-resolution computed tomography (CT) technology, a pixel resolution close to 1 µm allows for morphometry of microvessels comparable with classical light microscopy. CC µ-CT scanning provides new data from the microvascular tree that can be used both for the mathematical modeling (Debbaut et al, 2014) or quantitative purposes (Jiřík et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

The use of porcine corrosion casts for teaching human anatomy

Eberlová

Liška

Mírka

et al. 2017

Annals of Anatomy - Anatomischer Anzeiger

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In teaching and learning human anatomy, anatomical autopsy and prosected specimens have always been indispensable. However, alternative methods must often be used to demonstrate particularly delicate structures. Corrosion casting of porcine organs with Biodur E20 Plus is valuable for teaching and learning both gross anatomy and, uniquely, the micromorphology of cardiovascular, respiratory, digestive, and urogenital systems. Assessments of casts with a stereomicroscope and/or scanning electron microscope as well as highlighting cast structures using color coding help students to better understand how the structures that they have observed as two-dimensional images actually exist in three dimensions, and students found using the casts to be highly effective in their learning. Reconstructions of cast hollow structures from (micro-)computed tomography scans and videos facilitate detailed analyses of branching patterns and spatial arrangements in cast structures, aid in the understanding of clinically relevant structures and provide innovative visual aids. The casting protocol and teaching manual we offer can be adjusted to different technical capabilities and might also be found useful for veterinary or other biological science classes.

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Section: Introductionmentioning

confidence: 99%

The use of porcine corrosion casts for teaching human anatomy

Eberlová

Liška

Mírka

et al. 2017

Annals of Anatomy - Anatomischer Anzeiger

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“…; Jiřík et al. ). Moreover, quantitative data regarding the size and density of hepatocytes may also be combined with multi‐level modeling of hepatic perfusion (Debbaut et al.…”

Section: Discussionmentioning

confidence: 98%

Stereological analysis of size and density of hepatocytes in the porcine liver

et al. 2016

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The porcine liver is frequently used as a large animal model for verification of surgical techniques, as well as experimental therapies. Often, a histological evaluation is required that include measurements of the size, nuclearity or density of hepatocytes. Our aims were to assess the mean number-weighted volume of hepatocytes, the numerical density of hepatocytes, and the fraction of binuclear hepatocytes (BnHEP) in the porcine liver, and compare the distribution of these parameters among hepatic lobes and macroscopic regions of interest (ROIs) with different positions related to the liver vasculature. Using disector and nucleator as design-based stereological methods, the morphometry of hepatocytes was quantified in seven healthy piglets. The samples were obtained from all six hepatic lobes and three ROIs (peripheral, paracaval and paraportal) within each lobe. Histological sections (thickness 16 μm) of formalin-fixed paraffin-embedded material were stained with the periodic acid-Schiff reaction to indicate the cell outlines and were assessed in a series of 3-μm-thick optical sections. The mean number-weighted volume of mononuclear hepatocytes (MnHEP) in all samples was 3670 ± 805 μm (mean ± SD). The mean number-weighted volume of BnHEP was 7050 ± 2550 μm . The fraction of BnHEP was 4 ± 2%. The numerical density of all hepatocytes was 146 997 ± 15 738 cells mm of liver parenchyma. The porcine hepatic lobes contained hepatocytes of a comparable size, nuclearity and density. No significant differences were identified between the lobes. The peripheral ROIs of the hepatic lobes contained the largest MnHEP with the smallest numerical density. The distribution of a larger MnHEP was correlated with a larger volume of BnHEP and a smaller numerical density of all hepatocytes. Practical recommendations for designing studies that involve stereological evaluations of the size, nuclearity and density of hepatocytes in porcine liver are provided.

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“…The choice of the basic parameters to be quantified by our software generator was based on studies with real micro‐CT (Jiřík et al, ). This includes volume, surface, length, and number of objects, which are often expressed as relative densities calculated per ROI.…”

Section: Methodsmentioning

confidence: 99%

“…The three‐dimensional and quantitative nature of the micro‐CT makes it a tool of choice for estimating shrinkage of materials (Burey et al, ) and propagation of material fractures and cavitation (Neves, Coutinho, Alves, & de Assis, ). Some of the estimates are based on or fully compatible with stereological principles and spatial statistics (Baddeley & Jensen, ; Mouton, ; Stoyan, Kendall, & Mecke, ) and may be assessed automatically, provided the image data have a sufficient contrast (Jiřík et al, ; Mouton et al, ). However, surfaces are especially potentially sensitive to the settings of the micro‐CT scanning and further image postprocessing, such as resolution, noise, preferential spatial arrangement (anisotropy), filtration, and thresholding during binarization of grayscale images.…”

Section: Introductionmentioning

confidence: 99%

Generating standardized image data for testing and calibrating quantification of volumes, surfaces, lengths, and object counts in fibrous and porous materials using X‐ray microtomography

Jiřík

Bartoš

Tomášek

et al. 2018

Microscopy Res & Technique

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Quantification of the structure and composition of biomaterials using micro-CT requires image segmentation due to the low contrast and overlapping radioopacity of biological materials. The amount of bias introduced by segmentation procedures is generally unknown. We aim to develop software that generates three-dimensional models of fibrous and porous structures with known volumes, surfaces, lengths, and object counts in fibrous materials and to provide a software tool that calibrates quantitative micro-CT assessments. Virtual image stacks were generated using the newly developed software TeIGen, enabling the simulation of micro-CT scans of unconnected tubes, connected tubes, and porosities. A realistic noise generator was incorporated. Forty image stacks were evaluated using micro-CT, and the error between the true known and estimated data was quantified. Starting with geometric primitives, the error of the numerical estimation of surfaces and volumes was eliminated, thereby enabling the quantification of volumes and surfaces of colliding objects. Analysis of the sensitivity of the thresholding upon parameters of generated testing image sets revealed the effects of decreasing resolution and increasing noise on the accuracy of the micro-CT quantification. The size of the error increased with decreasing resolution when the voxel size exceeded 1/10 of the typical object size, which simulated the effect of the smallest details that could still be reliably quantified. Open-source software for calibrating quantitative micro-CT assessments by producing and saving virtually generated image data sets with known morphometric data was made freely available to researchers involved in morphometry of three-dimensional fibrillar and porous structures in micro-CT scans.

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Stereological quantification of microvessels using semiautomated evaluation of X-ray microtomography of hepatic vascular corrosion casts

Cited by 12 publications

References 40 publications

The use of porcine corrosion casts for teaching human anatomy

The use of porcine corrosion casts for teaching human anatomy

Stereological analysis of size and density of hepatocytes in the porcine liver

Generating standardized image data for testing and calibrating quantification of volumes, surfaces, lengths, and object counts in fibrous and porous materials using X‐ray microtomography

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