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We show that photosensitized phospholipid oxidation, initiated by the lipid-conjugated fluorophore TopFluor-PC, causes defects, namely, membrane tubes and vesicle-like structures, in supported lipid bilayers (SLBs). Lipid oxidation is detrimental to the integrity of the lipid molecules; when oxidized, they undergo a conformational expansion, which causes membrane tubes to protrude from the SLB. Lipid oxidation is verified by FT-IR spectroscopy, and area expansion is observed in Langmuir trough experiments. Upon growing to a critical length, the membrane tubes arising from SLBs rapidly undergo transition to vesicle-like structures. We find a correlation between the maximum tube length and the diameter of the resulting vesicle, suggesting the conservation of the surface area between these features. We use geometric modeling and the measured tube length and vesicle radius to calculate the tube radius; our calculated mean tube diameter of 243 nm is comparable to other groups' experimental findings. In the presence of fluid flow, membrane tubes can be extended to tens to hundreds of microns in length. SLBs composed of saturated lipids resist light-induced tubulation, and the inclusion of the lipophilic antioxidant α-tocopherol attenuates the tubulation process and increases the light intensity threshold for tubulation.

Fluorescent lipid probes are commonly used to label membranes of cells and model membranes like giant vesicles, liposomes, and supported lipid bilayers (SLB). Here, we show that excitation of fluorescent lipid probes with BODIPY-like conjugates results in a significant acceleration of the rupture and SLB formation process for unsaturated phospholipid vesicles on SiO 2 surfaces. The resulting SLBs also have smaller measured masses, which is indicative of a reduction in membrane thickness and/or membrane density. The excitation of fluorescent probes with NBD and Texas Red conjugates does not accelerate the SLB formation process. In the absence of fluorescent probes or light, the inclusion of oxidized phospholipids also accelerates SLB formation. The excitationinduced acceleration caused by BODIPY-like probes is eliminated when the probes are present with saturated phospholipids not susceptible to oxidation, and it is attenuated when a lipophilic antioxidant (α-tocopherol) is present. These results suggest that BODIPY−phospholipid conjugates are photosensitizers, and their excitation causes oxidation of lipid membranes, which significantly alters membrane properties.

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