Orca flash 2.8 scientific cmos camera

A DOPC/DOPG (1 : 2) or DOPC/DOPE/TOCL (2 : 1 : 1) mixture was dissolved in chloroform in a cylindrical container. A thin lipid film was prepared on the wall of the container as descried above. The lipid film was hydrated with 1 mM HEPES buffer, pH 6.5, containing 200 mM sucrose by placing the buffer gently on the lipid film at 30 °C and incubating overnight, resulting in the formation of GUVs (≥1 μm in diameter).
RHG (0.3 μM) was added to GUVs (total lipid, 500 μM) in 1 mM HEPES buffer, pH 6.5. The solution containing RHG and GUVs was incubated for at least 30 min to obtain uniform fluorescence intensities among different GUV surfaces. GUVs were imaged at room temperature using an Olympus IX 71 microscope (Olympus, Center Valley, PA, USA). An Olympus 60×/1.4 NA Plan Apo oil immersion lens was used as an objective lens. Excitation light was obtained using an Hg lamp with a U-MWIY2 filter set (Olympus; excitation wavelength, 545–580 nm). Microscopic images were recorded using an Orca-Flash2.8 Scientific CMOS Camera (Hamamatsu, Japan).