Sphingolipids have limited solubility in many common solvents such as chloroform, hexane, ethyl ether, and even methanol. To overcome this insolubility problem, various solvent mixtures have been developed. One of the most universal sphingolipid solvents is a mixture of chloroform/methanol/water. However, while this solvent system is very useful for analytical testing, it is very toxic towards cells and cannot be used in live cell cultures or other in vivo applications. Several alternative methods have been developed to provide sphingolipid solutions suitable for live-cell studies. Although these methods were primarily developed for ceramides and glucosylceramides, they may be able to be adapted for other lipids.