Abstract This study examined bryophyte community composition in relation to microsite and microenvironmental variation at different scales in three conifer-dominated stands in the boreal forest of Alberta, Canada. We documented bryophyte assemblage on specific microsite types (physiognomic forms providing substrates for moss colonization: logs, stumps, tree bases, undisturbed patches of forest floor, disturbed patches of forest floor), and at coarser scales: mesosites (625 m2 plots within stands), and stands (10 ha). Patterns of variation in bryophyte composition arising from the microsite sampling were clearly related to microsite type and, for woody substrates, to microsite quality (decay class; hardwood vs. softwood). Microenvironment (moisture, pH, temperature, light) also had some influence on bryophyte composition of woody microsite types. Forest floor moisture, pH, and light were related to bryophyte composition of undisturbed patches of forest floor while forest floor moisture and temperature were significant correlates for disturbed forest floor. At the coarser-scale, surface moisture and forest floor moisture were related to bryophyte assemblage of mesosites; this was partially reflective of differences among stands. We conclude that bryophyte species composition in these forests is related to a hierarchy of factors including fine scale variation in the type and quality of available microsites along with microenvironmental variation at different scales. Management efforts to preserve bryophyte biodiversity will need to incorporate this complexity.