Atlantic salmon populations are declining, and warming river temperatures in the summer months are thought to be a significant contributing factor. We describe the time course of cellular and metabolic responses to an ecologically relevant short-term thermal cycle in juvenile Atlantic salmon. We then examined whether this heat event would affect tolerance to a subsequent heat shock in terms of critical thermal maximum (CT max ). Fish induced heat shock protein 70 in red blood cells, heart, liver, and red and white muscle; whole blood glucose and lactate transiently increased during the heat cycle. In contrast, we observed no significant effect of a prior heat shock on CT max . The CT max was positively correlated with Fulton’s condition factor suggesting that fish with greater energy reserves are more thermally tolerant. Atlantic salmon activate cellular protection pathways in response to a single thermal cycle and appear to cope with this short-term, ∼1 d heat shock, but this challenge may compromise the ability to cope with subsequent heat events.