Introduction: Therapy of atopic dermatitis encloses use of medicated and nonmedicated preparations. Results of clinical and biophysical procedures indirectly describe the condition of the impaired skin barrier (SB). Direct evaluation of SB integrity is only possible by electron microscopical visualization, e.g. intercellular lipid lamellae (ICLL) organization of the stratum corneum. Method: SB integrity was measured by morphometric analysis of ICLL in healthy and atopic skin and after a 15-day treatment (plus 7-day follow-up) of atopic skin with a glycerin preparation. Results: Significant treatment effect was shown by the restoration of the ICLL. Conclusions: The study reveals that morphometric analysis of ICLL organization is suitable to differentiate between healthy and diseased skin and to semiquantitatively determine the effect of a nonmedicated glycerin formulation. Limitation: Small treatment cohort.