Behavioral markers are used in training and assessment of nontechnical skills (NTSs). This study tested the reliability of a unique set of markers designed to help assess heightened emotional activity (HEA) as a response to perceived threats. Using five markers that represent instances of negative emotional states, 27 commercial airline pilots recorded their observations of HEA in eight video vignettes of scripted flight deck situations. The raters’ scores were analyzed using Cronbach’s alpha for dichotomous data. The alpha statistics indicated high inter-rater reliability across all five HEAs. This study demonstrates that the chosen instances of HEA can be reliably identified by airline pilots.