Protocol for high-power, brain-focused microwave fixation to define rodent metabolism
Analysis of metabolites provides key insights into brain physiology and function. Due to post-mortem metabolism, both the euthanasia method and dissection time can make a critical difference. Here, we describe a protocol to euthanize rodents by microwave irradiation. This workflow details steps for animal placement, tissue fixation, and post-fixation processing. This protocol enables the rapid halting of metabolic activity for the accurate assessment of the metabolome in situ for analyses such as mass spectrometry and nuclear magnetic resonance. For complete details on the use and execution of this protocol, please refer to Juras et al.<sup>1</sup>.
Authors
- Alex R. Cantrell (ORCID: https://orcid.org/0000-0003-4134-9981)
- Tara R. Hawkinson (ORCID: https://orcid.org/0000-0002-8743-5503)
- Kia H. Markussen (ORCID: https://orcid.org/0000-0001-9446-0400)
- Lyndsay E.A. Young (ORCID: https://orcid.org/0000-0002-6873-5674)
- Bret N. Smith (ORCID: https://orcid.org/0000-0003-4743-1564)
- Joanna Long (ORCID: https://orcid.org/0000-0002-5572-6859)
- Ramon C. Sun (ORCID: https://orcid.org/0000-0002-3009-1850)
- Matthew S. Gentry (ORCID: https://orcid.org/0000-0001-5253-9049)
- Douglas Andres
- Roberto A. Ribas (ORCID: https://orcid.org/0009-0008-9024-1292)
- Jelena A. Juras (ORCID: https://orcid.org/0000-0002-2821-9069)
- Craig W. Vander Kooi (ORCID: https://orcid.org/0000-0002-7463-3914)
- James W. Collins (ORCID: https://orcid.org/0000-0002-6396-2579)
- Madison B. Webb (ORCID: https://orcid.org/0000-0003-4709-1723)
- Terrymar Medina
- Amelia A Bunnell
Institutions
- University of Kentucky (US)
- Medical University of South Carolina (US)
- University of Florida (US)
- Colorado State University (US)
- Florida College (US)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-04-25
- DOI
- https://doi.org/10.1016/j.xpro.2025.103794
- Citations
- 1
- Primary Topic
- Mitochondrial Function and Pathology
- Type
- article
- Field-Weighted Citation Impact
- 0.81
Funders
- National Science Foundation
- Kentucky Spinal Cord and Head Injury Research Trust
- National Institutes of Health
- Markey Cancer Center, University of Kentucky
- National Institute on Aging
- National Cancer Institute
- National Institute of Diabetes and Digestive and Kidney Diseases
- National Institute of Neurological Disorders and Stroke