How to dissect mouse stellate ganglion?

Tamara Buijs

Sure, no problem:

Materials:

4x needle or pin

1x scalpel blade

1x surgical tools

3x dish

1x tube with medium

1x cotton buds or kimtech wipes

1x syringe

1x pentaject

Aluminium foil

1. Kill mouse by cervical dislocation or pentobarbital injection

2. Spray with ethanol

3. Fix paws and head to table with needles

4. Cut open skin along midline of thorax using a scalpel and pin to table with needles

5. Separate ribs along midline via a subxiphoideal approach using scissors

6. Trim ribs using scissors

7. Remove lungs, heart, and all other organs until you see the spine.

8. Cut aorta and oesophagus.

9. Use Kimtech wipes to remove excess blood.

10. Stellate ganglion is located under 1st or 2nd rib near costovertebral joint, in close proximity to the spine. Next to the subclavian/mammalian artery. Look for star shape and translucent appearance. The stellate ganglia are whitish and look like small onions. Caudally connected to the paravertebral ganglia.

11. Follow paravertebral chain down to confirm it’s the stellate ganglion.

12. The stellate ganglion is the first ganglion larger than the others

13. The two arms that emerge from the stellate ganglion encircle the subclavian artery

Notes: to avoid a bloody chaos you may want to gravity perfuse the dead mouse first with ~10mL PBS via a needle in the left ventricular chamber. This will push out the blood from the vasculature and may give you a more clear view of the anatomy.

Chen Yu

I hope this message finds you well. Your interest in dissecting the mouse stellate ganglion is a critical endeavor in the study of autonomic nervous system functions and its implications in various physiological and pathological processes. The dissection of the stellate ganglion, due to its small size and proximity to vital structures, requires meticulous technique and precision. Below is a detailed, step-by-step guide designed to assist you in this intricate procedure.

Preparation:

Equipment and Setting Preparation: Ensure all dissection tools are sterilized and sharp. Set up a well-illuminated, comfortable workspace with a dissecting microscope. Prepare a cold PBS (Phosphate Buffered Saline) solution to rinse and store the ganglia after dissection.

Animal Preparation: Following institutional guidelines and ethical standards for animal research, euthanize the mouse using an approved method. Place the mouse in a supine position and disinfect the surgical area with 70% ethanol to minimize contamination.

Dissection Steps:

Initial Incision and Exposure:Make a midline incision from the lower jaw down to the chest to expose the throat and chest area. Use retractors to gently hold the incision open. Carefully dissect and retract the salivary glands and surrounding connective tissues to expose the trachea and esophagus.

Identification of Landmark Structures:Locate the trachea and gently move it aside to reveal the carotid artery, which is a critical landmark for identifying the stellate ganglion. The stellate ganglion in mice is located at the cervical-thoracic junction, where the sympathetic chain converges with the first thoracic nerve.

Stellate Ganglion Dissection:With fine dissection tools, carefully isolate the stellate ganglion. It is usually found adjacent to the vertebral column, at the level of the first rib, and may require careful dissection of surrounding tissues for clear visualization. Once the stellate ganglion is identified, gently separate it from the surrounding tissue using micro-dissection scissors and forceps. It is crucial to minimize damage to the ganglion and adjacent structures.

Ganglion Removal:After isolating the ganglion, use fine forceps to lift it slightly and cut the connecting nerves and blood vessels with micro-scissors. Ensure that the ganglion is completely freed from surrounding tissues. Immediately place the dissected ganglion into cold PBS to maintain tissue integrity until further processing.

Post-Dissection:

Tissue Processing: Depending on the subsequent analysis, the stellate ganglion can be processed for histological examination, gene expression analysis, or other biochemical assays.
Equipment Cleanup: Sterilize all dissection tools and clean the workspace to prevent contamination and ensure safety.

Tips for Success:

Practice on non-essential specimens to refine your dissection skills before attempting on valuable samples.
Maintain patience and precision throughout the dissection process, as rushing can damage the delicate ganglion or surrounding structures.
Consider recording the dissection under the microscope for educational purposes or to review your technique.

This guide aims to provide a foundational framework for the dissection of the mouse stellate ganglion. Adaptations may be necessary based on specific experimental requirements or anatomical variations.

Should you have any further questions or require additional assistance, please do not hesitate to contact me. I am here to support your scientific exploration and endeavors.

Best regards,

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