When evaluating the effect of husbandry and biomethodologies on the well-being of laboratory mice, it is critical to utilize measurements that allow the distinguishing of acute stress from chronic stress. One of the most common measurements of stress in laboratory animals is the corticosterone assessment. However, while this measurement provides a highly accurate reflection of the animal's response to acute stressors, its interpretation is more prone to error when evaluating the effect of chronic stress. This study evaluated the use of the neutrophil:lymphocyte (NE:LY) ratio as an assessment of chronic stress in male and female C57Bl/6N mice as compared to serum corticosterone. One group of mice was exposed to mild daily stressors for 7 days, while the control group was handled with normal husbandry. The NE:LY ratio and serum corticosterone levels were significantly elevated in the chronically stressed mice, though a significant increase in corticosterone was only significant in males when compared by sex. The chronically stressed mice also demonstrated significantly fewer entries into the open arms and less time spent in the open arms of the elevated plus maze, suggesting that the mild daily stressors had induced a state of distress. The findings of this study confirm that the NE:LY ratio is a valid measurement for chronic stress in the laboratory mouse. However, these assays do not distinguish between distress or eustress, so behavioral and physiological assessments should always be included to determine a complete assessment of the well-being of the mouse.