Amphid neurons were stained with DiI (D-282, Molecular Probes) as described (45) with slight modifications. Briefly, well-fed young adult animals were collected from 6-cm plates, washed twice with M9 buffer, and incubated in 1 ml of M9 buffer containing 5 μl of DiI [dilution of 1:200 of stock (2 mg/ml)] overnight at room temperature. After incubation, the animals were washed three times with M9 buffer and transferred to NGM plates seeded with E. coli OP50. After incubating for 1 hour, the animals were mounted on agarose pads with 50 mM sodium azide and imaged using a Zeiss M2 microscope with a mechanized stage, Zen software, and Texas red filters.

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