Protocol for high-power, brain-focused microwave fixation to define rodent metabolism

Alex R. Cantrell; Tara R. Hawkinson; Jelena A. Juras; Madison B. Webb; Terrymar Medina; Roberto A. Ribas; James Collins; Amelia A. Bunnell; Lyndsay E.A. Young; Kia H. Markussen; Douglas A. Andres; Joanna R. Long; Craig W. Vander Kooi; Bret N. Smith; Ramon C. Sun; Matthew S. Gentry

doi:10.1016/j.xpro.2025.103794

Protocol for high-power, brain-focused microwave fixation to define rodent metabolism

Alex R. Cantrell, Tara R. Hawkinson, Jelena A. Juras, Madison B. Webb, Terrymar Medina, Roberto A. Ribas, James Collins, Amelia A. Bunnell, Lyndsay E.A. Young, Kia H. Markussen, Douglas A. Andres, Joanna R. Long, Craig W. Vander Kooi, Bret N. Smith, Ramon C. Sun, Matthew S. Gentry

Research output: Contribution to journal › Article › peer-review

Abstract

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.¹

Original language	English
Article number	103794
Journal	STAR Protocols
Volume	6
Issue number	2
DOIs	https://doi.org/10.1016/j.xpro.2025.103794
State	Published - Jun 20 2025

Bibliographical note

Publisher Copyright:
© 2025 The Authors

Funding

This study was supported by the National Institute of Health (NIH) grants R01AG066653 , R01CA266004 , R01AG078702 , RM1NS133593 , and R01CA288696 to R.C.S. and R35NS116824 and R33NS111081 to M.S.G.; NIH grants R01DK122811 and R01NS092552 to B.N.S.; and NIH NS102196 and Kentucky Spinal Cord and Head Injury Research Trust 22-1A to D.A.A. L.E.A.Y. was supported by NIH/ NCI F99CA264165 . This research was also supported by funding from the University of Kentucky Markey Cancer Center and the NIH-funded Biospecimen Procurement and Translational Pathology Shared Resource Facility, as well as the Cancer Research Informatics Shared Resource Facility , of the University of Kentucky Markey Cancer Center ( P30CA177558 ). A portion of this work was performed in the McKnight Brain Institute at the National High Magnetic Field Laboratory\u2019s Advanced Magnetic Resonance Imaging and Spectroscopy (AMRIS) Facility, which is supported by National Science Foundation Cooperative Agreement DMR-2128556 and the State of Florida.

Funders	Funder number
University of Kentucky Markey Cancer Center
Florida AandM University and Florida State University
National Institutes of Health (NIH)	R01AG078702, R01NS092552, P30CA177558, R33NS111081, R35NS116824, R01CA288696, R01DK122811, NS102196, R01CA266004, R01AG066653, RM1NS133593
National Institutes of Health (NIH)
National Childhood Cancer Registry – National Cancer Institute	F99CA264165
National Childhood Cancer Registry – National Cancer Institute
U.S. Department of Energy Chinese Academy of Sciences Guangzhou Municipal Science and Technology Project Oak Ridge National Laboratory Extreme Science and Engineering Discovery Environment National Science Foundation National Energy Research Scientific Computing Center National Natural Science Foundation of China	DMR-2128556
U.S. Department of Energy Chinese Academy of Sciences Guangzhou Municipal Science and Technology Project Oak Ridge National Laboratory Extreme Science and Engineering Discovery Environment National Science Foundation National Energy Research Scientific Computing Center National Natural Science Foundation of China

Keywords

metabolism
metabolomics
neuroscience

ASJC Scopus subject areas

General Neuroscience
General Biochemistry, Genetics and Molecular Biology
General Immunology and Microbiology

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10.1016/j.xpro.2025.103794

Cite this

Cantrell, A. R., Hawkinson, T. R., Juras, J. A., Webb, M. B., Medina, T., Ribas, R. A., Collins, J., Bunnell, A. A., Young, L. E. A., Markussen, K. H., Andres, D. A., Long, J. R., Vander Kooi, C. W., Smith, B. N., Sun, R. C., & Gentry, M. S. (2025). Protocol for high-power, brain-focused microwave fixation to define rodent metabolism. STAR Protocols, 6(2), Article 103794. https://doi.org/10.1016/j.xpro.2025.103794

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title = "Protocol for high-power, brain-focused microwave fixation to define rodent metabolism",

abstract = "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.1",

keywords = "metabolism, metabolomics, neuroscience",

author = "Cantrell, \{Alex R.\} and Hawkinson, \{Tara R.\} and Juras, \{Jelena A.\} and Webb, \{Madison B.\} and Terrymar Medina and Ribas, \{Roberto A.\} and James Collins and Bunnell, \{Amelia A.\} and Young, \{Lyndsay E.A.\} and Markussen, \{Kia H.\} and Andres, \{Douglas A.\} and Long, \{Joanna R.\} and \{Vander Kooi\}, \{Craig W.\} and Smith, \{Bret N.\} and Sun, \{Ramon C.\} and Gentry, \{Matthew S.\}",

note = "Publisher Copyright: {\textcopyright} 2025 The Authors",

year = "2025",

month = jun,

day = "20",

doi = "10.1016/j.xpro.2025.103794",

language = "English",

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Cantrell, AR, Hawkinson, TR, Juras, JA, Webb, MB, Medina, T, Ribas, RA, Collins, J, Bunnell, AA, Young, LEA, Markussen, KH, Andres, DA, Long, JR, Vander Kooi, CW, Smith, BN, Sun, RC & Gentry, MS 2025, 'Protocol for high-power, brain-focused microwave fixation to define rodent metabolism', STAR Protocols, vol. 6, no. 2, 103794. https://doi.org/10.1016/j.xpro.2025.103794

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AU - Cantrell, Alex R.

AU - Hawkinson, Tara R.

AU - Juras, Jelena A.

AU - Webb, Madison B.

AU - Medina, Terrymar

AU - Ribas, Roberto A.

AU - Collins, James

AU - Bunnell, Amelia A.

AU - Young, Lyndsay E.A.

AU - Markussen, Kia H.

AU - Andres, Douglas A.

AU - Long, Joanna R.

AU - Vander Kooi, Craig W.

AU - Smith, Bret N.

AU - Sun, Ramon C.

AU - Gentry, Matthew S.

PY - 2025/6/20

Y1 - 2025/6/20

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AB - 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.1

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Protocol for high-power, brain-focused microwave fixation to define rodent metabolism

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

Access to Document

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