Pressure Fixation of Isolated Murine Pulmonary Valve: A Procedure to Isolate and Chemically Fix the Mouse Pulmonary Valve in Closed Conformation
Published: April 30, 2023
Abstract
Source: Liu, Y., et al. Surgery and Sample Processing for Correlative Imaging of the Murine Pulmonary Valve. J. Vis. Exp. (2021)
In this video, we describe the procedure to isolate and chemically fix the mouse pulmonary value in closed conformation via perfusion pressurization.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Pulmonary valve excision Autoclave the necessary tools needed for the mouse dissection. This includes fine scissors, micro forceps, micro vascular clamps, clamp applying forceps, microneedle holders, spring scissors, and retractors. Acclimate all mice for a minimum of 2 weeks before the operation. Remove C57BL…
Disclosures
The authors have nothing to disclose.
Materials
| 25% glutaraldehyde (aq) | EMS | 16210 | Primary fixative component |
| 0.9% sodium chloride injection | Hospira Inc. | NDC 0409-4888-10 | |
| 1 mL syringe | BD | 309659 | |
| 10 mL syringe | BD | 309604 | |
| 22G needle | BD | 305156 | |
| 3-way stopcock | Smiths Medical ASD, Inc | MX5311L | |
| 4% paraformaldehyde (aq) | EMS | 157-4-100 | Primary fixative component |
| C57BL/6 mice | Jackson Laboratories | 664 | Approximately 1 yo |
| Clamp applying forcep | FST | 00072-14 | |
| Dumont #5 forcep | FST | 11251-20 | |
| Dumont #5/45 forceps | FST | 11251-35 | |
| Dumont #7 fine forcep | FST | 11274-20 | |
| Fine scissor | FST | 14028-10 | |
| Saline solution (sterile 0.9% sodium chloride) | Hospira Inc | NDC 0409-0138-22 | |
| Surgical microscope | Leica | M80 | |
| Tish needle holder/forcep | Micrins | MI1540 | |
| Trimmer | Wahl | 9854-500 | |
| Black polyamide monofilament suture, 10-0 | AROSurgical instruments Corporation | TI38402 | |
| Black polyamide monofilament suture, 6-0 | AROSurgical instruments Corporation | SN-1956 | |
| Ketamine hydrochloride injection | Hospira Inc | NDC 0409-2053 | |
| L-aspartic acid | Sigma-Aldrich | 56-84-8 | Staining component |
| Lead nitrate | EMS | 17900 | Staining component |
| Sodium cacodylate trihydrate | Sigma-Aldrich | 6131-99-3 | Buffer |
| Non-woven songes | McKesson Corp. | 94442000 | |
| Micro-adson forcep | FST | 11018-12 | |
| Millex-GP filter, 0.22 um, PES 33mm, non-sterile | EMD Millipore | SLGP033NS |
Tags
Cite This Article
Pressure Fixation of Isolated Murine Pulmonary Valve: A Procedure to Isolate and Chemically Fix the Mouse Pulmonary Valve in Closed Conformation. J. Vis. Exp. (Pending Publication), e20801, doi: (2023).