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Simultaneous Three-Dimensional Vascular and Tubular Imaging of Whole Mouse Kidneys With X-ray μCT

Published online by Cambridge University Press:  06 July 2020

Willy Kuo Open the ORCID record for Willy Kuo [Opens in a new window] ,
Ngoc An Le Open the ORCID record for Ngoc An Le [Opens in a new window] ,
Bernhard Spingler Open the ORCID record for Bernhard Spingler [Opens in a new window] ,
Roland H. Wenger Open the ORCID record for Roland H. Wenger [Opens in a new window] ,
Anja Kipar Open the ORCID record for Anja Kipar [Opens in a new window] ,
Udo Hetzel Open the ORCID record for Udo Hetzel [Opens in a new window] ,
Georg Schulz Open the ORCID record for Georg Schulz [Opens in a new window] ,
Bert Müller Open the ORCID record for Bert Müller [Opens in a new window]  and
Vartan Kurtcuoglu Open the ORCID record for Vartan Kurtcuoglu [Opens in a new window]
Willy Kuo
Affiliation:
University of Zurich, Institute of Physiology, Winterthurerstrasse 190, 8057Zurich, Switzerland University of Zurich, National Centre of Competence in Research, Kidney. CH, Winterthurerstrasse 190, 8057Zurich, Switzerland University of Basel, Biomaterials Science Center, Department of Biomedical Engineering, Gewerbestrasse 14, 4123Allschwil, Switzerland
Ngoc An Le
Affiliation:
University of Zurich, Department of Chemistry, Winterthurerstrasse 190, 8057Zurich, Switzerland
Bernhard Spingler
Affiliation:
University of Zurich, Department of Chemistry, Winterthurerstrasse 190, 8057Zurich, Switzerland
Roland H. Wenger
Affiliation:
University of Zurich, Institute of Physiology, Winterthurerstrasse 190, 8057Zurich, Switzerland University of Zurich, National Centre of Competence in Research, Kidney. CH, Winterthurerstrasse 190, 8057Zurich, Switzerland
Anja Kipar
Affiliation:
University of Zurich, Laboratory for Animal Model Pathology (LAMP), Institute of Veterinary Pathology, Vetsuisse Faculty, Winterthurerstrasse 268, 8057Zurich, Switzerland
Udo Hetzel
Affiliation:
University of Zurich, Electron Microscopy Unit, Institute of Veterinary Pathology, Vetsuisse Faculty, Winterthurerstrasse 268, 8057Zurich, Switzerland
Georg Schulz
Affiliation:
University of Basel, Biomaterials Science Center, Department of Biomedical Engineering, Gewerbestrasse 14, 4123Allschwil, Switzerland
Bert Müller
Affiliation:
University of Basel, Biomaterials Science Center, Department of Biomedical Engineering, Gewerbestrasse 14, 4123Allschwil, Switzerland
Vartan Kurtcuoglu*
Affiliation:
University of Zurich, Institute of Physiology, Winterthurerstrasse 190, 8057Zurich, Switzerland University of Zurich, National Centre of Competence in Research, Kidney. CH, Winterthurerstrasse 190, 8057Zurich, Switzerland University of Zurich, Zurich Center for Integrative Human Physiology, 8057Zurich, Switzerland
*
*Author for correspondence: Vartan Kurtcuoglu, E-mail: [email protected]
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Abstract

Concurrent three-dimensional imaging of the renal vascular and tubular systems on the whole-kidney scale with capillary level resolution is labor-intensive and technically difficult. Approaches based on vascular corrosion casting and X-ray micro computed tomography (μCT), for example, suffer from vascular filling artifacts and necessitate imaging with an additional modality to acquire tubules. In this work, we report on a new sample preparation, image acquisition, and quantification protocol for simultaneous vascular and tubular μCT imaging of whole, uncorroded mouse kidneys. The protocol consists of vascular perfusion with the water-soluble, aldehyde-fixable, polymeric X-ray contrast agent XlinCA, followed by laboratory-source μCT imaging and structural analysis using the freely available Fiji/ImageJ software. We achieved consistent filling of the entire capillary bed and staining of the tubules in the cortex and outer medulla. After imaging at isotropic voxel sizes of 3.3 and 4.4 μm, we segmented vascular and tubular systems and quantified luminal volumes, surface areas, diffusion distances, and vessel path lengths. This protocol permits the analysis of vascular and tubular parameters with higher reliability than vascular corrosion casting, less labor than serial sectioning and leaves tissue intact for subsequent histological examination with light and electron microscopy.

Keywords

computed tomographycontrast agentskidneytissue stainingvascular imaging
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 26 , Issue 4 , August 2020 , pp. 731 - 740
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Copyright
Copyright © Microscopy Society of America 2020

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Footnotes

a

These authors contributed equally to the present study.

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