Biliary System Resin Casting: A Technique for Analyzing 3D Architecture of Biliary System in Murine Model Using Radiopaque Resin Injection
Published: April 30, 2023
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 Preparation 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. Prepare 1 mL aliquots of diluted resin per mouse. Yellow Resin: Dilute yellow silicone rubber (hig…
Representative 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 of the tubing is stretched and cut at an angle to create a beveled tip. The ruler in A and B is a centimeter ruler, with major increments of 1 cm, i…
Declarações
The authors have nothing to disclose.
Materials
| 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 |
Tags
Citar este artigo
Biliary System Resin Casting: A Technique for Analyzing 3D Architecture of Biliary System in Murine Model Using Radiopaque Resin Injection. J. Vis. Exp. (Pending Publication), e20788, doi: (2023).