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Protocol for high-power, brain-focused microwave fixation to define rodent metabolism
Alex R Cantrell1, Tara R Hawkinson1, Jelena A Juras2
1Department of Biochemistry and Molecular Biology, University of Florida, College of Medicine, Gainesville, FL 32610, USA.
STAR Protocols
|April 27, 2025
Summary
This study presents a microwave irradiation protocol for euthanizing rodents, rapidly halting metabolic activity. This method ensures accurate in situ metabolome assessment for techniques like mass spectrometry.
Area of Science:
- Neuroscience
- Metabolomics
- Biochemistry
Background:
- Metabolite analysis offers critical insights into brain physiology and function.
- Post-mortem metabolism, influenced by euthanasia and dissection timing, can significantly alter results.
- Accurate metabolomic assessment requires rapid preservation of in situ metabolic states.
Purpose of the Study:
- To describe a protocol for rodent euthanasia using microwave irradiation.
- To detail a workflow for rapid metabolic activity halting and tissue fixation.
- To enable accurate in situ metabolome analysis.
Main Methods:
- Rodent euthanasia via microwave irradiation.
- Detailed protocol for animal placement and tissue fixation.
- Post-fixation processing steps for sample preparation.
Main Results:
- The microwave irradiation protocol effectively halts metabolic activity in rodent tissues.
- This method allows for the preservation of the in situ metabolome.
- Facilitates downstream analyses such as mass spectrometry and nuclear magnetic resonance.
Conclusions:
- Microwave irradiation provides a rapid and effective method for rodent euthanasia.
- This protocol is crucial for accurate metabolomic studies of the brain.
- Enables reliable in situ assessment of the metabolome, minimizing post-mortem artifacts.
