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"So, how was work today?"

Let me see if I can remember how yesterday went. This is so I can remember the insanity some day in the future.

8:30 am: Arrive at lab.
Check scintillation counter for yesterday's results. Write them down, then wash the scintillation trays so they will be dry in time for use later in the day.
Go to walk-in cooler and centrifuge sample set 1 (first TCA precipitation; 24 samples in each of the numbered sets I reference)
Put sample set 2 in scintillation tray and put in scintillation counter to count radioactivity (protein samples from 4-5 days ago, stored in drawer)
Add scintillation fluid to sample set 3 (lipid samples from yesterday) and put in drawer to dark-adapt for at least 1 hour
Pull styrofoam cooler out of plastic cooler inside the walk-in cooler and bring to main lab space
Write out labels for the 24 crickets to be injected on a set of thick-walled glass tubes and a set of 25-mL Erlenmeyer flasks
Put tubes in styrofoam cooler, walk up five stories and one building over to get dry ice, then put styrofoam cooler back in plastic cooler back in walk-in cooler
Prepare 8 more corks for the flasks (folding and pinning filter paper to the corks; didn't have enough dry the night before to prepare all of them in advance)
Inject crickets with radiolabeled acetate (around an hour and change of nonstop motion, averaging ~2 minutes between injections, plus additional prep work between sets of 8 crickets to make controls and keep crickets at 27 degrees C in one of the cricket rooms) - they then incubate in the flasks for 3 hours when acetate-injected
Retrieve sample set 1 from the walk-in cooler, remove TCA with a pulled pipette and discard; add second batch of TCA, vortex, and put back in the walk-in cooler
Get final weights for food dishes and frass dishes from yesterday's 21 crickets (these have to sit out in ambient conditions for 24 hours before being re-weighed)
Pick frass out of the food dishes for the crickets that were just injected and put it in 1-ounce deli cups to sit out; leave food dishes to sit out, too. Think the same thought every day: if I ever switch careers and try to get a job as a cake decorator, I am totally going to list "picking insect frass out of food with fine forceps" as relevant experience.
Set up 24 scintillation vials to collect the filter paper strips from the crickets

Eat lunch for 20 minutes

Grab first set of 8 crickets from the cricket room, grab styrofoam cooler
Every two minutes: pop off a cork, shake out a cricket, lift the lid to the styrofoam cooler and stuff her into one of the freezing-cold tubes and then cap the tube. Then use a pair of forceps to strip the filter paper off the pins and stuff it into a scintillation vial. To the vial add scintillation fluid, cap, label, and put in a tray.
In the one-minute increments between crickets, work on other incremental tasks: pour sample set 4 (yesterday's protein samples) into scintillation vials, add scintillation fluid, cap, number, and store in drawer to dark-adapt and count two days later
Centrifuge sample set 1 again, remove the second batch of TCA, then add sodium hydroxide and tissue solubilizer and put back in the walk-in cooler until there's enough time to sonicate them
Pull out another set of 24 cricket ovarioles from the freezer to thaw and prep for lipid extraction, and write them down in my lab notebook (sample set 4)
…Crickets done. Go to scintillation counter, write down results from sample set 2, put sample set 3 plus the freshly collected filter paper samples (carbon dioxide) into the counter and start to count (lipid tray gets counted first so the carbon dioxide samples have time to dark-adapt while sitting in the scintillation counter). Bring sample set 2 back to the lab and stick back in a drawer for storage.
Okay, time to start the day's lipid samples. I guess that's sample set 5? They have been marinating in chloroform-methanol in the walk-in cooler for ~24 hours. They get sonicated, then stuck in the centrifuge. While they are centrifuging, I sonicate sample set 1 and stick it on the heat block, caps open.
Remove chloroform-methanol from sample set 5 with a pulled pipette and transfer to a clean set of eppendorf tubes (contains extracted lipids). Tedious.
Add more chloroform-methanol to sample set 5, sonicate again, and centrifuge again. Meanwhile, add the first bit of chloroform-methanol to sample set 4 so they can marinate overnight
…several more detailed steps to purify the chloroform-extracted lipids from sample set 5. These samples wind up in scintillation vials and are left in the fume hood overnight so all of the chloroform-methanol will evaporate out (it interferes with detection of radioactivity). Then leave the lipid-free remnants in the fume hood to dry out for 1-2 hours. At some point I close the caps to sample set 1 so they can cook overnight without drying out. Also pull out the first set of 8 crickets (now dead) so they can thaw out for dissection.
Dissect the 24 crickets: cut open, pin in place, remove ovarioles and weigh them, note the flight muscle characteristics. When finished, store in the freezer for now.
Add TCA to the lipid-free remnants from sample set 5, sonicate, and leave in the walk-in cooler overnight. Phew, done in there, it gets COLD after a while at 4 degrees Celsius.
Go to cricket room and sort crickets: remove all of the winged adults from the 12 focal aquaria and either kill them or move them to a new aquarium to be kept as breeders. Keep track of how many long-winged vs. short-winged females I remove.
Double-check current sample sizes for different treatments (especially how many long-winged crickets with pink flight muscle). Portion out fresh diets for the crickets that I will sort out and set up tomorrow (diets need to sit out in ambient conditions for 24 hours before given to crickets) - these will be ready to inject next Monday.
Wash the radioactive labware
Wash the cricket cages from the day's crickets so they'll be ready for tomorrow
6:30 pm: Leave lab and hurry to bike shop before it closes at 7 pm.

It would be interesting to wear a pedometer on days like today. Also, I am SO RELIEVED to have discovered that my old bike shoes (Specialized) are actually shaped right to take the pressure off of my metatarsals/metatarsalgia.

Tomorrow in addition to repeating the above I will need to also: prepare fresh radiolabel, prepare a full set of corks, and set up crickets on the diets. I'll be relieved if I get done before 8 pm tomorrow.



( 3 remarks — Remark )
Sep. 17th, 2015 03:29 am (UTC)
Frass frass frass.
New word for the day.
Sep. 22nd, 2015 02:35 pm (UTC)
So, I read this paper a long time ago, where researchers put caterpillars onto different kinds of plants in a greenhouse and then measured what the caterpillars ate, and how much frass they made.

They collected up the frass from underneath the plants using an aspirator, but in the paper they referred to it as a "frasspirator." I was so pleased that they managed to slip the word into the paper.
Sep. 23rd, 2015 01:31 am (UTC)
I bet the whole department was giggling over that. I love that.
( 3 remarks — Remark )

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