To apply accurate and uniform osmotic pressures to liposomes, they can be formed using the spontaneous transfer method in Solutions With different osmolarities. The majority of liposomes unexpectedly opened large holes (several micrometers in diameter) in response to the osmotic pressure regardless of its strength, that is, the difference between the outside and inside solute (sucrose or KCl) concentrations. However, the lag time for any response, including the opening of it hole, after the formation of the liposome decreased with increasing osmotic pressure.