Oleosin Emulsion Dynamics in DOPC Monolayer

Oleosin emulsions consisting of 2.8 μL sucrose solution (with 3.92 μg or 0.28 μg oleosin) containing 100 μM resorufin and 60 μL mineral oil was added to the DOPC monolayer and centrifuged at 5,200 × g for 10 min at room temperature. Ten microliters of vesicle solution were collected from the tube. Ten microliters of 500 μM or 30 μM 16:0 lyso-PC micelles containing 0.14 mol % TopFluor lyso-PC was added to 10 μL of the lipid–oleosin vesicle solution. Time-lapse images were acquired using a CLSM. The TopFluor lyso-PC fluorescence and resorufin fluorescence were observed using a CLSM with a 60× oil-immersion lens using a diode laser (473 nm for TopFluor) and (559 nm for resorufin), respectively. The area of the vesicle was measured using a feel hand tool of ImageJ. The fluorescence of TopFluor on these vesicles was measured using ImageJ.