The first few days working with dii staining I had a few issues with the entire worm being fluorescent making it impossible to actually see the amphid neurons. This was just caused by the dye clumping at the bottom of the centrifuge tube and was easily solved by making sure I mixed the solution better. The other difficulty came with identifying exactly which neuron was which. This was difficult because almost every worm I looked at was oriented differently on the slide and sometimes all the neurons looked too clustered to accurately identify.
I have looked at a decent amount of these worms now and from what I’ve observed there is not a difference in expression between males and hermaphrodites.
I have also still been working on getting a successful cross between my him-5:dmd-6::GFP and him-5:AWA::mCherry worms. I’ve tried numerous times and still haven’t had any success. I’m observing a few more crosses today that I set up so hopefully I’ll have some luck.