Help Needed

Hello everyone! I am in desperate need of some recommendations.

I am not an expert on C. elegans. In fact, I have never worked with them. However, because of my interest in physiology and metabolism,
I decided to take on a senior independent study project where I study C. elegans lifespan in response to various stress conditions, specifically focusing on
certain genetic knockouts. I have prepared extensively for this project as part of my school’s independent study program, doing the necessary research
on my specific topic as well as studying protocols. However, as I have said, I have never worked with them formally. Near the end of this past semester,
I tried worm transfer a few times (unsuccessfully) and learned how to prepare, chunk and maintain OP50 plates. Through this, I realized just how much
practice I will need before working efficiently with the organism, specifically in relation to the whole scoring process, and I became generally anxious
about the prospect of the coming year.

This anxiety has reached a peak this summer, considering new positive, non-academic developments in my life that I wish to work towards during this coming school
year, combined with the prospect of balancing a tough class schedule AND this independent study project.

My question is, how tough will this year be for me? I know you all are biased towards C. elegans research, but think back to when you began: Will I truly
be able to pursue other aspects of life while getting adequate sleep (6 hours a night will do) as someone quickly learning to work with C. elegans and then
diving headfirst into a project?

Most importantly, what sort of a daily time commitment should I expect? I plan to score perhaps once every two days during each assay, and I will need
to do maintenance procedures, of course, a few times a week if I am not mistaken. I am dedicated to whatever task is at hand in my life, and I am
more than willing to put in extra hours at the beginning of the year to really get comfortable with the organism.

Please be honest with me. I am genuinely considering pursuing another project simply because science is probably not going to be my eventual direction in
life. Thus, I don’t want to sacrifice my life for this project. Also, if you think this coming year is feasible for a hardworking, disciplined college student,
tips for success would be much appreciated.

Thank you all so much.

I think a lot depends on resources, reagents, and ambitions. I don’t think “learning to use C. elegans” should be a major bar if you’re just maintaining strains and checking whether worms are still alive. An hour or two a day (every other day if you trust FUdR) to do a moderate number of lifespan experiments doesn’t sound unreasonable (not that I have done a lot of lifespan experiments), and maintaining a few strains can be just a few minutes every day or two. But: will you have reliable access to a decent dissecting scope (one that lets you see pharyngeal pumping and doesn’t give you a headache after an hour of using it) and a good constant-temperature incubator? Where are you getting your plates from? You can hand-pour your plates, but I’d estimate that at about 300 plates per hour of actual pouring, and that’s after mixing up the media (~20 minutes) and almost three hours spent autoclaving the media and waiting for it to cool. You can get studying done during that time, but you can’t leave the lab for more than an hour. Do you have someone knowledgeable you can talk to about whether your experiments are worthwhile, and whether they’re novel?

I’m not particularly worried about plate pouring or worm maintenance. I’m worried about basically anything that requires directly handling the worms (transfer, scoring, egg transfer) because of the dexterity it requires. I’m afraid that the learning curve may be too steep for someone who has a lot of other stuff to do (I’m not getting my PhD, so this won’t be my sole focus). What is your or someone else’s impression of the learning curve required for these procedures? Any other opinions on how this will work out for me?

I think that work with C. elegans is much easier compared to other laboratory animals (among other things worms do not bite unlike mice :slight_smile: ).
The most difficult for me was transfer of a single animal with a worm picker. After almost one hour of arduous work
I had approximately 5 worms transferred (naturally all of them dead :frowning: ), it was very frustrating. However that was just at the start.
After few days of trying you can easily master this skill.
I have been working with worms for over three years now and so far have not encountered more difficult thing than worm picker manipulating.
Scoring may be a bit boring and tiring (after hours of staring in microscope and hundreds of animals checked).
It is very difficult to estimate time you are likely to spend in the lab without deeper knowledge of the study you are going to conduct.
However I wold not be too concerned, after fist few frustrating days you shoul be able to work with worms easily.

Good luck.

Thanks for your post! So say I’m scoring a total of 108 wells for single worms every other day. How long would you expect that to take in a single sitting once I’ve gotten the hang of it? Really can’t emphasize enough how helpful these responses are.

as others have already said, the answer to your question regarding how much time would be needed to complete your project depends on a few important factors.

Transferring/Picking worms: chunking worms from one plate to another takes up very little time, but you have to have everything you need there to hand and hence, a dedicated space in the lab where you can work and store a few things such as a small bottle of alcohol, spirit burner, scalpel etc.
You also need plates that have an overnight growth of the bacteria (so set up the day before).

Picking (using a fine platinum shovel to move worms from one plate to another) takes practice, all of us have buried worms into the agar learning this technique.

Scoring worms as dead/alive is also time consuming when you first start doing it, but, as with all things, practice makes one better and more efficient.

For me, the main question regarding your project is not whether you can master these techniques or not, we all have to a greater or lesser extent. My main concern (and perhaps yours to consider) is how easy it would be to run such a project with the facilities you have available.

As Hillel has already mentioned, having an thermostatically-controlled incubator is a must for your lifespan assays, our lab has no air conditioning and temperatures vary according to season by more than 5 degrees. If your worms are subject to different temperatures on different days and/or between different experiments, then your data is not worth much.

Having a dedicated space to work is also important, rather than chasing around each time for pipettes, plates, reagents etc.

Finally, without knowing what you want to do exactly, commenting on the scientific feasibility/value of your project is a bit difficult…but you need to make sure that someone who knows what would be involved looks through what you are proposing so that you don’t waste your time pursuing a project that yields very little.

Balancing social life, academic lectures etc. and a final year project is hard…most of have been through this…I wouldn’t even say that the project you do needs to be your life’s work, but you need to work as if it was for a few months and then move on to where you really want to go with your life.


Lately I have been scoring animals for QL neuroblast descendants position along
anterior/posterior axis (neuroblasts with mec-7::gfp to visualise them).
I conducted RNAi knock-down with 5 different RNA molecules+1 control and watched how position of neuroblast descendants changed.
Scoring was done under fluorescent microscope (over 100 animals per each RNAi counted, cca 700 in total).
Overall time spent was about 3 hours (included is time needed for washing plates, spining down worms, transfer to agarose pad with NaN3).
You can proceed pretty fast once you know what to look for.
I have no experience with life span studies whatsoever so I do not even know what you are checking and how long that can take.
As for equipment it is nicely described in earlier comments (by steveh and HillelSchwartz)

Do a small pilot trial with just one strain (~ 50 animals, no treatment). Record the amount of time it takes for prep (plates, media, etc) and for doing the actual experiment (worm handling, scoring lethality). That will give you an approximate realistic idea of the time requirements as well as the potential issues and pitfalls.

Thank you all for your replies! I’ve sorted out my basic work space and have access to all necessary tools (microscope, autoclave, incubator, etc. etc.). I’m doing an experiment where I subject genetic knockouts of arginine kinase isoforms to various stress conditions (variable pH, heat shock etc.) and score every couple of days for lifespan. My only pressing worry is the learning curve of scoring. Age synchronization, worm transfer, and not killing the little guy with my wire seem all to have a pretty steep learning curve and to consume a great deal of time. Correct me if I’m wrong?
I appreciate these responses, and honestly if I can be confident in the feasibility of fitting this into my life, I am excited to perform the experiment, as I have pretty much all experimental details laid out. Thanks in advance for any more thoughts.