Indirect Calorimetry to Assess Energy Balance in Mice: Measurement and Data Analysis

Understanding the factors affecting body weight regulation requires careful measurement of food intake and metabolic rates. Modern indirect calorimetry systems are designed to record these features. Here, we describe our approach for reproducible analysis of energy balance experiments performed using indirect calorimetry. CalR, a free online web tool, calculates both instantaneous and cumulative totals for metabolic variables including food intake, energy expenditure, and energy balance making it an excellent start for analyzing energy balance experiments. Energy balance may be one of the most important metrics that CalR calculates as it provides a clear picture of metabolic trends resulting from experimental interventions. Because of the complexity of indirect calorimetry devices and the frequency of mechanical breakdowns, we place a heavy emphasis on the importance of data refinement and visualization. Plots representing energy intake or energy expenditure versus body mass or physical activity can help to identify a malfunctioning apparatus. We also introduce a critical visualization of experimental quality control: a plot of the change in energy balance versus the change in body mass, which simultaneously represents many of the essential components of indirect calorimetry. These analyses and data visualizations allow the investigator to make inferences about experimental quality control and the validity of experimental results.