Removal of vertical forest structure (overstory canopy, understory, wood debris, and forest floor layers) from forest harvesting and subsequent silviculture practices modify incoming radiation and interception of precipitation. These changes affect near-surface and belowground microclimatic conditions which are key drivers to other ecosystem processes such as seedling establishment, nutrient availability, carbon sequestration, and biodiversity. This study quantified the effect of the removal of vertical structure on bi-weekly and seasonally-integrated temperature (air and soil), vapour pressure deficit (VPD), frost events, and soil moisture over seven years at the Island Lake Biomass Harvesting Trial in northeastern Ontario. Environmental conditions have not recovered since harvest and were differentially affected by removing different strata. For air temperature, successive removal of strata had an additive effect of increasing air temperature. For soil temperature, the response was the same, but the removal of the canopy and forest floor produced the largest increases in the soil temperature. For VPD, removal of the canopy caused increases in VPD, while forest floor removal resulted in decreases, with the understory and woody debris layers having little effect. A similar pattern was observed for frost events and soil moisture. Tracking bi-weekly changes in conditions showed that subtle shifts could occur in the early and late growing season, that were not captured by the annual integrated metrics. These results provide a foundational element to help interpret changes in other ecological processes and patterns following biomass harvesting that will, in turn, help establish improved best management practices for sustainable forest management.