Towards Automated Three-Dimensional Tracking of Nephrons through Stacked Histological Image Sets

Bhikha, Charita; Andreasen, Arne; Christensen, Erik; Letts, Robyn F. R.; Pantanowitz, Adam; Rubin, David M.; Thomsen, Jesper Skovhus; Zhai, Xuguang

doi:10.1155/2015/545809

Cited by 3 publications

(1 citation statement)

References 19 publications

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“…For instance, an optical-microscopy-based 3D reconstruction was developed, in which the 3D structure was reconstructed by serial sectioning from about a thousand consecutive 2.5 μm-thick 2D images of a resin-embedded mouse kidney so that a single nephron could be segmented manually [ 6 , 7 ]. This reconstruction method was successfully applied to confirm the urine concentrating mechanism of rat renal inner medulla [ 8 , 9 ]; efforts toward automated segmentation are in progress [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

Compact laboratory-based X-ray microscope enabling nondestructive 3D structure acquisition of mouse nephron with high speed and better user accessibility

et al. 2022

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X-ray microscopes adopting computed tomography (CT) enable nondestructive 3D visualization of biological specimens at micron-level resolution without conventional 2D serial sectioning that is a destructive/laborious method and is routinely used for analyzing renal biopsy in clinical diagnosis of kidney diseases. Here we applied a compact commercial system of laboratory-based X-ray microscope to observe a resin-embedded osmium-stained 1 mm strip of a mouse kidney piece as a model of renal biopsy, toward more efficient diagnosis of kidney diseases. A reconstructed CT image from several hours of data collection using CCD detector allowed us to unambiguously segment a single nephron connected to a renal corpuscle, which was consistent with previous reports using serial sectioning. Histogram analysis on the segmented nephron confirmed that the proximal and distal tubules were distinguishable on the basis of their X-ray opacities. A 3D rendering model of the segmented nephron visualized a convoluted structure of renal tubules neighboring the renal corpuscle and a branched structure of efferent arterioles. Furthermore, another data collection using sCMOS detector with much shorter data acquisition time of 15 min provided similar results from the same samples. These results suggest a potential application of the compact laboratory-based X-ray microscope to analyze mouse renal biopsy.

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Section: Introductionmentioning

confidence: 99%

Compact laboratory-based X-ray microscope enabling nondestructive 3D structure acquisition of mouse nephron with high speed and better user accessibility

et al. 2022

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Nephron morphometry in mice and rats using tomographic microscopy

Letts

Zhai

Bhikha

et al. 2017

American Journal of Physiology-Renal Physiology

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The aim was to quantify the glomerular capillary surface area, the segmental tubular radius, length, and area of single nephrons in mouse and rat kidneys. Multiple 2.5-µm-thick serial Epon sections were obtained from three mouse and three rat kidneys for three-dimensional reconstruction of the nephron tubules. Micrographs were aligned for each kidney, and 359 nephrons were traced and their segments localized. Thirty mouse and thirty rat nephrons were selected for further investigation. The luminal radius of each segment was determined by two methods. The luminal surface area was estimated from the radius and length of each segment. High-resolution micrographs were recorded for five rat glomeruli, and the capillary surface area determined. The capillary volume and surface area were corrected for glomerular shrinkage. A positive correlation was found between glomerular capillary area and proximal tubule area. The thickest part of the nephron, i.e., the proximal tubule, was followed by the thinnest part of the nephron, i.e., the descending thin limb, and the diameters of the seven identified nephron segments share the same rank in the two species. The radius and length measurements from mouse and rat nephrons generally share the same pattern; rat tubular radius-to-mouse tubular radius ratio ≈ 1.47, and rat tubular length-to-mouse tubular length ratio ≈ 2.29, suggesting relatively longer tubules in the rat. The detailed tables of mouse and rat glomerular capillary area and segmental radius, length, and area values may be used to enhance understanding of the associated physiology, including existing steady-state models of the urine-concentrating mechanism.

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Three-dimensional visualization of the mouse renal connecting tubule

Deng

Miao

et al. 2018

Digital Medicine

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Background and Objectives: In this study, the spatial courses of the connecting tubule (CNT) of nephron from different depth of cortex and the collecting duct (CD) in mouse kidney was established with the aid of three-dimensional visualization technology. Subjects and Methods: Kidneys from three C57/BL/6J mice were removed after perfusion fixation. The tissue blocks were cut perpendicular to the longitudinal axis of the kidney and embedded in Epon-812. A total of 2000, 2.5-μm-thick consecutive sections were obtained from the renal capsule to papilla. After acquiring the digitalized images and alignment, the CNT from 137 nephrons were traced with the custom-made programs. The spatial arrangement of the CNT was visualized, and the length was measured. Results: Each CD received CNT from 5 to 7 nephrons. The CNT from different level of cortex all drained into the CD at superficial cortex but took different path. The CNT from superficial cortical nephron joined CD directly, while the CNT from middle and juxtamedullary cortical nephron joined each other to form an arcade, and the latter drained into CD at superficial cortex. The length of the arcade ranged 400-800 μm. Conclusions: The CNT joined CD at superficial cortex, which means the fluid along the CD from the cortex would not be added from outside, contributing to stabilization of the hormone regulation in the transportation of solutes and water along the CD from cortex to medulla.

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Towards Automated Three-Dimensional Tracking of Nephrons through Stacked Histological Image Sets

Cited by 3 publications

References 19 publications

Compact laboratory-based X-ray microscope enabling nondestructive 3D structure acquisition of mouse nephron with high speed and better user accessibility

Compact laboratory-based X-ray microscope enabling nondestructive 3D structure acquisition of mouse nephron with high speed and better user accessibility

Nephron morphometry in mice and rats using tomographic microscopy

Three-dimensional visualization of the mouse renal connecting tubule

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