Palmitic Acid, continued
Oct. 13th, 2015 08:37 pm![[personal profile]](https://www.dreamwidth.org/img/silk/identity/user.png){kind=small}
rebeccmeisterI 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.
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.
