Surgery and Sample Processing for Correlative Imaging of the Murine Pulmonary Valve

Yifei Liu; Yong-Ung Lee; Tai Yi; Ken Wu; Cedric Bouchet-Marquis; Han Chan; Christopher K. Breuer; David W. McComb

doi:10.3791/62581

Surgery and Sample Processing for Correlative Imaging of the Murine Pulmonary Valve

JoVE Journal
Engineering

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JoVE Journal Engineering

Surgery and Sample Processing for Correlative Imaging of the Murine Pulmonary Valve

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07:34 min

August 5, 2021

DOI: 10.3791/62581-v

Yifei Liu¹, Yong-Ung Lee², Tai Yi², Ken Wu³, Cedric Bouchet-Marquis³, Han Chan³, Christopher K. Breuer², David W. McComb¹

¹Center for Electron Microscopy and Analysis, The Department of Materials Science and Engineering,Ohio State University, ²Center for Regenerative Medicine,Nationwide Children’s Hospital, ³Thermo Fisher Scientific

Overview

This protocol outlines a correlative workflow for the excision, pressurization, fixation, and imaging of the murine pulmonary valve. It aims to elucidate the gross conformation and local extracellular matrix structures.

Key Study Components

Area of Science

Neuroscience
Biology
Regenerative Medicine

Background

The murine pulmonary valve presents challenges due to its inherent heterogeneity.
Understanding its structure-function relationship is crucial for various biomedical applications.
Controlled fixation and imaging techniques are essential for accurate structural analysis.
This study utilizes high-resolution imaging to map structures at multiple length scales.

Purpose of Study

To develop a reliable protocol for studying the murine pulmonary valve.
To capture detailed images of the valve's structure.
To correlate high-resolution images with precise anatomical locations.

Methods Used

Autoclaving dissection tools for mouse preparation.
Euthanizing and positioning the mouse for thoracotomy.
Performing excision and fixation of the pulmonary valve.
Utilizing correlative imaging techniques for structural analysis.

Main Results

Successful capture of the pulmonary valve's structure at various scales.
Demonstration of the protocol by Dr. Tai Yi.
Mapping of local high-resolution images to anatomical locations.
Insights into the extracellular matrix structures of the valve.

Conclusions

The developed workflow is effective for studying the murine pulmonary valve.
High-resolution imaging provides valuable insights into valve structure.
This approach can enhance understanding of valve function and pathology.

Frequently Asked Questions

What is the significance of this protocol?

It addresses the challenges in studying the murine pulmonary valve's structure-function relationship.

Who demonstrates the procedure?

Dr. Tai Yi, the Director of Microsurgery at Nationwide Children's Hospital.

What tools are required for the dissection?

Fine scissors, microforceps, microvascular clamps, and other specialized tools.

How is the mouse prepared for dissection?

The mouse is euthanized and positioned in a dorsal recumbent position.

What imaging techniques are used?

Correlative imaging techniques are employed to capture detailed structural information.

What are the expected outcomes of this study?

To gain insights into the structure and function of the murine pulmonary valve.

Here, we describe a correlative workflow for the excision, pressurization, fixation, and imaging of the murine pulmonary valve to determine the gross conformation and local extracellular matrix structures.

This protocol is significant because it's designed to answer questions about the structure function correlate of the murine pulmonary valve, which is generally tricky because of its inherent heterogeneity. The controlled fixation and correlative imaging were used to capture the structure on multiple length scales. Local high-resolution images can then be mapped back to the precise anatomical location on the pulmonary valve.

Demonstrating the procedure will be Dr.Tai Yi, the Director of Microsurgery in the Center of Regenerative Medicine at Nationwide Children's Hospital. Begin by autoclaving the tools needed for the mouse dissection, including fine scissors, microforceps, microvascular clamps, clamp applying forceps, microneedle holders, spring scissors, and retractors. After euthanizing an adult C57BL/6 mouse, place it in a dorsal recumbent position on a tray, secure its limbs with tape and perform the thoracotomy.

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