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.¹.