The objective of the investigations performed was to determine the way various optical isomers of α-tocopherol affect the permeability of lipid membrane – a simple model of the cell membrane. Owing to its hydrophobic properties, α-tocopherol is easy soluble in the lipid core of cell membrane, and this phenomenon impacts the structure, stability, and other properties of the cell membrane. D-α-tocopherol is a natural form of vitamin E, which is also present in plant oils and vegetables. And the DL-α-tocopherol is a synthetic vitamin E commonly used as a food additive. Some authors claim that, despite the same chemical structure of the two forms of vitamin E, the synthetic vitamin E may have a different effect in vivo compared with the natural form of this vitamin. Thus, it is important to identify mechanisms of interactions ensuing among α-tocopherol, other biologically significant molecules, and cell structures on a molecular level. In the investigations carried out, the D-α-tocopherol and mixture of D and L-α-tocopherol (α-T) were applied. The liposomes (lipid membranes) were made from the phosphatidyl choline dipalmitate (DPPC) and the L-α-phosphatidyl choline (PC) of egg yolk. A fluorescence probe method with an 8-anilino-1-naphthalene-sulfonic acid (ANS) as a colouring dye was applied to monitor structural changes occurring in the lipid membrane. The process of ANS probe penetrating the DPPC membrane being in a gel phase occurred by immediate absorption (it lasted between one to several seconds), with no adsorption phase present. When α-tocopherol was added to the membrane, the penetration rate of the probe did not change. The D-α-tocopherol was the only case when the amount of absorbed ANS was 25% of the value corresponding with the pure DPPC membrane and with DPPC with DL-α-tocopherol. In the DPPC membrane, a significant increase in the membrane permeability was stated. It was attributed to the crystalline transition. In the case of DPPC with D-α- and DL-α-tocopherol added, it was noted that the maximal increase in the membrane permeability due to the change in the membrane phase was about 60% lower than in the case of DPPC alone, and the changes in the membrane permeability as observed in the temperature of the main crystalline transition, were characterized by a milder course. α-tocopherol inhibited the process of the ANS probe penetrating the membrane in a liquid crystalline phase. No significant differences were found in the membrane permeability in the presence of D and DL-α-tocopherol.