We evaluated macroinvertebrate-based metrics from Hester–Dendy samplers (HDs) that were either new or previously deployed (HS status) and were processed with either picking or scraping methods. The metrics assessed included the total number of individuals, the number of intolerant individuals, the total number of families, and the number of Ephemeroptera, Plecoptera, Trichoptera (EPT) individuals. No significant differences in mean number of individuals, intolerant individuals, and total families were detected between processing techniques or HD status (all p > 0.05). Mean number of EPT individuals did not differ between processing methods (p > 0.05), but previously deployed HDs yielded significantly more EPT individuals than new HDs (p  =  0.001). The EPT data are of particular interest because these organisms are important components of indices of biotic integrity and are used to assess water-quality status. Previously deployed and new HDs should not be used in the same study because colonization by EPT organisms differs between them.