DUCT: Double Resin Casting followed by Micro-Computed Tomography for 3D Liver Analysis

被引：2

作者：

Hankeova, Simona ^{[1
]}

Salplachta, Jakub ^{[2
]}

Van Hul, Noemi ^{[1
]}

Kavkova, Michaela ^{[2
]}

Iqbal, Afshan ^{[1
]}

Zikmund, Tomas ^{[2
]}

Kaiser, Jozef ^{[2
]}

Andersson, Emma R. ^{[1
]}

机构：

[1] Karolinska Inst, Stockholm, Sweden

[2] Cent European Inst Technol BUT, Brno, Czech Republic

来源：

JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2021年 / 175期

基金：

瑞典研究理事会;

关键词：

INTRAHEPATIC BILE-DUCTS; BILIARY SYSTEM;

D O I：

10.3791/62941

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

The liver is the biggest internal organ in humans and mice, and high auto-fluorescence presents a significant challenge for assessing the three-dimensional (3D) architecture of the organ at the whole-organ level. Liver architecture is characterized by multiple branching lumenized structures, which can be filled with resin, including vascular and biliary trees, establishing a highly stereotyped pattern in the otherwise hepatocyte-rich parenchyma. This protocol describes the pipeline for performing double resin casting micro-computed tomography, or "DUCT". DUCT entails injecting the portal vein and common bile duct with two different radiopaque synthetic resins, followed by tissue fixation. Quality control by clearing one lobe, or the entire liver, with an optical clearing agent, allows for pre-screening of suitably injected samples. In the second part of the DUCT pipeline, a lobe or the whole liver can be used for micro-computed tomography (microCT) scanning, (semi-)automated segmentation, and 3D rendering of the portal venous and biliary networks. MicroCT results in 3D coordinate data for the two resins allowing for qualitative as well as quantitative analysis of the two systems and their spatial relationship. DUCT can be applied to postnatal and adult mouse liver and can be further extended to other tubular networks, for example, vascular networks and airways in the lungs.

引用

页数：19