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

Author:

Kunishima Naoki1ORCID,Takeda Yoshihiro1,Hirose Raita1,Kume Satoshi23,Maeda Mitsuyo4,Oguchi Akiko56,Yanagita Motoko67,Shibuya Hirotoshi8,Tamura Masaru8,Kataoka Yosky49,Murakawa Yasuhiro5710,Ito Koichiro11,Omote Kazuhiko1

Affiliation:

1. X-ray Research Laboratory, Rigaku Corporation , Akishima, Tokyo 196-8666, Japan

2. Laboratory for Pathophysiological and Health Science, RIKEN Center for Biosystems Dynamics Research , Kobe, Hyogo 650-0047, Japan

3. Center for Health Science Innovation, Osaka City University , Osaka 530-0011, Japan

4. Multi-Modal Microstructure Analysis Unit, RIKEN-JEOL Collaboration Center , Kobe, Hyogo 650-0047, Japan

5. RIKEN Center for Integrative Medical Sciences , Yokohama, Kanagawa 230-0045, Japan

6. Department of Nephrology, Graduate School of Medicine, Kyoto University , Kyoto 606-8501, Japan

7. Institute for the Advanced Study of Human Biology (ASHBi), Kyoto University , Kyoto 606-8501, Japan

8. Technology and Development Team for Mouse Phenotype Analysis, RIKEN BioResource Research Center , Tsukuba, Ibaraki 305-0074, Japan

9. Laboratory for Cellular Function Imaging, RIKEN Center for Biosystems Dynamics Research , Kobe, Hyogo 650-0047, Japan

10. IFOM―the FIRC Institute of Molecular Oncology , Milan 20139, Italy

11. New Market Development Office, Rigaku Corporation , Akishima, Tokyo 196-8666, Japan

Abstract

Abstract X-ray microscopes adopting computed tomography 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 a more efficient diagnosis of kidney diseases. A reconstructed computed tomography 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 scientific complementary metal-oxide semiconductor detector with a 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.

Funder

JSPS KAKENHI

RIKEN Aging Project

RIKEN Engineering Network Project

Publisher

Oxford University Press (OUP)

Subject

Radiology, Nuclear Medicine and imaging,Instrumentation,Structural Biology

Reference41 articles.

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