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Abstract In vitro grown plants may have abnormal structural and physiological features. However, the type of the sealing material used in tissue culture may minimize such abnormalities. This study evaluates the influence of the type of sealing of test tubes on the anatomical and ultrastructural features of leaves of Aechemea bromeliifolia (Rudge) Baker (Bromeliaceae), an ornamental bromeliad native to Brazil, grown in vitro. Three types of sealing were used: rigid polypropylene cap (PC), polyvinyl chloride film (PVC), and PC coupled with a microporous membrane (PM). Seedlings germinated in a greenhouse were also studied for comparison. Plants grown in test tubes sealed with PM were more similar to those from the greenhouse, as far as the pattern of stomatal opening, the presence of starch grains, and the organization of the internal membrane system of the chloroplasts is concerned. Plants cultivated in test tubes sealed with PC had higher stomatal density and the chloroplasts had large areas without thylakoids in the stroma. Plants grown in test tubes sealed with PVC had few or no starch grains. These results suggest that microporous membrane used coupled with PC sealing provided natural ventilation, thus contributing to a better plant development.