Hello again,
I'm glad to see you are pursuing this project. Heather has some valuable information, so I think between the both of us we should be able to assist you when needed.
I highly recommend you do more research on the care of the worms, their environment, their life cycle; many of your questions can be answered through research. Check out some of the resources I listed toward to the bottom of this message.
Yes the eggs and small worms (should they hatch) will need to be removed and put into their own petri dishes or discarded. I would suggest saving offspring and using them as an additional control group--just remember that if you choose to do that, label the petri dishes (i.e. eggs collected on day 1
from generation L1 could be labeled, "L2, day 1"). I highly suggest having more than 1 control group, as well as experimentals. I would say use at least 2 controls and experimentals (ideally 3). Many of your worms will die (usually not by the experimenter's fault)...so you want to make sure your sample size is abundant. I have had to restart colonies because the worms died...good thing they are always producing!
Here are some answers to your questions:
1. Since you will not know the ages of the first generation (L1), unless you receive them as eggs, I would wait until several hatch (L2) and begin your experiment with those. Use about 5-10 worms per plate, in case some die.
2. How do I separate the progeny or what is the best way to prevent mixing of adult and progeny? Do I have to check on them everyday and manually remove them? How do I identify the adults from the progeny?
You don't really need to separate males from hermaphrodites, it shouldn't affect your results (hermaphrodites have thin, whip-like tails; while males have blunt tails...see pictures in the link below). I'm not sure if you will need to or want to separate the sexes out or not, but if you do, let me know and I can explain that. Check your dishes
daily for eggs and hatched worms (they will be smaller and thinner than adults, and will move very quickly) and remove them. On that note, if your worms are getting sluggish/ not moving, they are going to die soon (they live only a couple of weeks). Also, since you will be transferring a known quantity of worms into each plate, you will know how many are new by the number of worms.
Question 2. 2. How do I separate the progeny or what is the best way to prevent mixing of adult and progeny? Do I have to check on them everyday and manually remove them? How do I identify the adults from the progeny?
How to remove L2 worms (and subsequent generations). You will be working under a dissecting microscope and using aseptic technique, you will need a Bunsen burner to heat your "inoculating loop" before scraping into your agar to retrieve new worms. Careful to let loop cool, so as to not singe the worms.
Question 3: You don't need any particular proportion; hermaphrodites self-fertilizing, and all offspring are clones (identical genetics). There will be very few males, as Heather stated.
Please check out these resources for information on worm care and anatomy.
Here is a site about how to grow them:
http://www.wormclassroom.org/c-elegans-cultivation
Here is an anatomical picture of worms:
http://www.wormatlas.org/ver1/handbook/anatomyintro/anatomyintro.htm
One important thing to mention is to label your dishes, or else your results will be very confusing later. Your labeling can include:
1. the generation (if you are keeping more than one, L1, L2, etc.)
2. Date of addition of worms onto dish.
3. Temperature worms are kept at.
4. your initials (if you aren't the only one working in the lab)
Let me know if this information is too confusing or you need further explanation.