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Palmitic Acid, continued

I keep having the thought: if I just work all the time, I won't have to worry about missing out on hobbies or activities with friends because soon enough I won't have any!

That's being extreme and melodramatic.

Yesterday, after I used up all of the radiolabel that I'd prepared, there was still residue left on the vial. So I added some ethanol, then removed a couple of small aliquots, which I put into scintillation vials and counted the radioactivity. Sure enough, there was still a bunch of the undissolved radiolabel left in the vial. But when I tried mixing in more phosphate-buffered saline and BSA, the palmitic acid refused to go into solution nicely. Upon further consultation with my boss, I heated it up and then sonicated it for 3 quick seconds. Well, that appeared to work, I concluded. I left work just after 9 pm, came home, ate dinner (big vat of mac-n-cheese with carrots and peas), tried to digest a little, and went to sleep.

This morning, I got up, had some breakfast, and got to work by 8 am. I had 63 crickets lined up for poking - 15 with acetate, 48 with palmitate. Trouble is, I only have 53 Erlenmeyer flasks and rubber stoppers. My solution was going to be tricky to manage, but it worked:

9 am - 9:30 am: Inject the acetate crickets
9:30 - 10: Finish preparations for remaining injections
10 - 11:40: Inject as many crickets as I can with remaining supplies (that's what, 38 crickets in an hour and 40 minutes? Not bad)
11:50 - 12: Shove lunch in my gullet
12 - 12:40: Wrap up the acetate crickets (freeze them, put their filter paper samples in scintillation vials). In the 60 seconds between crickets, wash, rinse, and towel-dry their flasks and corks
12:40 - 1: Inject more palmitate crickets. In this case, I only wound up doing 4 more injections because 6 of the long-winged crickets had already histolyzed their flight muscles, based on this trick I have been practicing where I look through part of the cuticle under the dissecting scope.
1 - 2:40: Wrap up the first set of palmitate crickets, as above. In the 60 seconds between crickets, work away at miscellaneous other projects or try to rest a little from the constant running.
2:40 - 3:40: Work on dissecting the acetate crickets and early palmitate crickets to verify flight muscle status and weigh ovarioles
3:40 - 4:00: Wrap up the second set of palmitate crickets.
4:00 - 5:45: Finish dissecting the remaining crickets
5:45 - 6:45: Cricket room time. Go through the 16 aquaria of juvenile crickets to pre-sort and remove all of the adults that have emerged over the past two days, so the aquaria will only contain Day 0 adults the next day (tomorrow I will sort out those Day 0 adults and set them up on our experimental diets so they will be ready to inject 5 days later).

So, all told, I injected 57 crickets today, and I managed to leave work before dark. Also, today's injections produced useable data.

Tomorrow I have a potential 73 crickets total. Following the same scheme as today, I could accommodate up to 68 crickets, which should be adequate. It might be helpful if I can get myself to work even earlier than 8 am, to ensure adequate and unhurried prep time.

I think my previous record number of crickets injected was somewhere around 34 crickets in one day, but with a radiolabel with a 2-hour time window instead of palmitate's 3-hour time window. In some ways, the 2-hour time window would be much simpler to work with, because these experiments have to occur during the timeframe between 9 am and 4 pm in order to avoid complications associated with hormonal differences between the long-winged and short-winged morphs that occur later in the day.

I am not going to manage to apply for that job that has an October 15 deadline, am I. Remember, the academic hiring cycle has a long timeframe. This is the period when universities advertise jobs that start a year from now - sometimes two years, depending on particulars. Plus out of courtesy I need to give my letter-writers adequate time to do their jobs.

At least I managed to get a preliminary illustration turned in for this caterpillar freelancing project and am starting to remember how to get Illustrator to do my bidding.



( 8 remarks — Remark )
Oct. 14th, 2015 05:09 am (UTC)
In high intensity mode, I see. As a member of your cheering squad, may your efforts yield all the abundance of usable data you can ever hope for. Goooooooo Rebecca!
Oct. 15th, 2015 02:11 am (UTC)
That sounds like an utterly grueling day.

Do you freeze the crickets to keep them still during injection or just to kill them and preserve them for assays? My experiments along these lines said that if I slowly froze grasshoppers they could withstand at least ten freeze/thaw cycles while still behaving in a more or less grasshopper-like manner.
Oct. 15th, 2015 02:16 am (UTC)
I just immobilize them in my hand when injecting them. Faster and less risky than trying to chill them. The freezing at the end is to kill them quickly so most of their tissues are relatively intact for various tissue extraction things.

At some point I really want to make a brief video about the whole process because I think it would be cool to share with people. Maybe not on a day when I need to inject this many crickets.
Oct. 15th, 2015 02:28 am (UTC)
Either that or a very, very brief video: about thirty seconds long.
Oct. 16th, 2015 02:24 pm (UTC)
Modern humans have such short attention spans, but I can see reasons in favor of this. Maybe two: How to Make a Radioactive Cricket, and then Why Make Radioactive Crickets.
Oct. 17th, 2015 01:02 am (UTC)
The moment I put on my black hat, I immediately think why NOT make radioactive crickets? Why not make a ton of them?
Oct. 17th, 2015 03:28 am (UTC)
I have been trying, hard, to turn into Cricket Woman, by repeatedly poking myself with radioactive cricket juices. It never works!!
Oct. 17th, 2015 04:54 am (UTC)
Well, that's the most real-life Kafka concept I have ever heard. Radioactive cricket woman!
Okay perhaps that's not working, but it may be a cure for chirpes!
( 8 remarks — Remark )

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