Biliary System Resin Casting: A Technique for Analyzing 3D Architecture of Biliary System in Murine Model Using Radiopaque Resin Injection

Abstract

Source: Hankeova, S. et al. DUCT: Double Resin Casting followed by Micro-Computed Tomography for 3D Liver Analysis. J. Vis. Exp. (2021)

This video describes the procedure for biliary system resin casting, for understanding 3D biliary architecture. This technique provides insights into the internal diameter and network connectivity of the biliary system, which could be used for liver regeneration analysis.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Double resin injections

1. Preparation
   1. Prepare resin solution. For double system resin injections, prepare both yellow and green resins as described in the steps 1.1.2-1.1.4.
   2. Prepare 1 mL aliquots of diluted resin per mouse.
   3. Yellow Resin: Dilute yellow silicone rubber (high radiopacity) with the clear diluent in a 3:1 dilution to prepare yellow resin for injection.
   4. Green Resin: Mix blue silicone rubber (undet....

Results

Figure 1: Injection set for resin casting. (A) Injection set #1 comprises a 30 G needle and PE10 tubing that is ~30 cm long. Injection set #2 is composed of a 23 G needle and PE50 tubing ~30 cm long. (B) The tip .......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
1.5 mL SafeSeal micro tubes  	Sarstedt	72.706
23 G butterfly needle with tubing 	BD bioscience	367283
23 G needle  	BD bioscience	305892
30 G needle	 BD bioscience	305106
Agarose  	Top-Bio	P045
Benzyl alcohol	Sigma Aldrich	108006
Benzyl benzoate 	 Sigma Aldrich	B6630
Corning 50 mL tubes  	Sigma Aldrich	CLS430829-500EA	 polypropylene
Cotton swabs 	Medicarier 	60406
Dissection Microscope  	Leica Camera AG	Leica M60
Dulbecco's phosphate-buffered saline  	ThermoFisher Scientific	14190144
Ethanol 70%  	VWR	83801.41
Falcon tube 15 mL 	Verkon	 331.850.084.006
Forceps curved 	Fine Science Tools	 11051-10	 Fine Graefe 10 cm curved
Forceps straight 	Fine Science Tools	  11050-10	Fine Graefe 10 cm straight
Formaldehyde solution 	 Sigma Aldrich	F8775
Hanks' Balanced Salt Solution 	ThermoFisher Scientific	14025092
Heparin	 Leo Pharma 	B01AB01	 5000 IE/mL
Isolfurane 	Baxter 	FDG9623
Methanol  	ThermoFisher Scientific	11413413
MICROFIL   	Flowtech	MV-122	synthetic resin yellow
MICROFIL  	Flowtech	MV-120 	synthetic resin blue
MICROFIL 	Flowtech	MV-diluent	  clear resin diluent
Pasteur pipette 	Verkon 	130.690.424.503
Peristaltic pump  	AgnThos	010.6131.M20
Rocker 	VWR 	444-0142
Silk suture  	AgnThos	14757	Black silk, 4-0, sterile, 100 m
Skin scissor  	 Fine Science Tools	14058-09	Iris straight tip 9 cm
Spring scissor   	Fine Science Tools	15000-03	Vannas micro, straight tip 2 mm
Syringe 1 mL Luer 	 BD bioscience	303172
Tubing PE10 	BD bioscience	427401
Tubing PE50 	BD bioscience 	427411