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

Area of Science:

• Anatomy
• Medical Imaging
• Biotechnology

Background:

• High autofluorescence in liver tissue hinders 3D architectural assessment at the whole-organ level.
• Liver structure is defined by intricate, branching vascular and biliary trees within the hepatocyte-rich parenchyma.

Purpose of the Study:

• To present a novel protocol, double resin casting micro-computed tomography (DUCT), for high-resolution 3D imaging of liver architecture.
• To enable detailed visualization and analysis of the liver's vascular and biliary systems.

Main Methods:

• The DUCT protocol involves dual radiopaque resin injection into the portal vein and common bile duct, followed by tissue fixation.
• Optical clearing allows for pre-screening of samples before micro-computed tomography (microCT) scanning.
• MicroCT scanning, segmentation, and 3D rendering are used to analyze the vascular and biliary networks.

Main Results:

• DUCT successfully visualizes the 3D architecture of both the portal venous and biliary networks within the liver.
• The method provides 3D coordinate data for detailed qualitative and quantitative analysis of these systems and their spatial relationships.
• The protocol is applicable to both postnatal and adult mouse livers.

Conclusions:

• DUCT is an effective method for overcoming autofluorescence challenges in whole-organ liver imaging.
• This technique allows for comprehensive analysis of the liver's complex vascular and biliary architecture.
• The DUCT pipeline can be adapted for studying other tubular networks, such as pulmonary vasculature and airways.