Identify a protein null mutant by sequencing alleles or performing western blots on extracts of mutants. This mutant can be used as a negative control for staining. A general note on handling worms without losing them. This protocol involves many transfers and washes of the worms. A common problem of beginning stainers is to lose some worms at each step, and thus end up with none at the end of the procedure.
Worms stick to plastic pipetteman tips unless detergent is present.
Thus the first transfers described below must be done with a glass pasteur pipette, but plastic pipetteman tips can be used for the later transfers when the worms are in buffers containing triton. I use a Shelton Scientific model VS-13. After spinning remove most of the liquid with a 1 ml pipetteman blue tip, and the remaining small amount of liquid with a small tip. Costar makes some white platic tips with a very small orifice that are good for removing liquid without losing worms. Fixing the worms.