So I get in this morning, and as I'm about to do my thing, these guys come in and start setting up. They tell me that they're fixing our hood and that once they set up and run the machine, no one will be able to enter the room for a couple of hours. I have a very important appointment in the City at 2 PM, so I'm like, nooooooo no no no, I didn't come in early to lab to push everything back by a few hours.
No one told me that they were going to do maintenance on the hood. Good thing I came in before the guys started! My project is SUPER time sensitive--I have
to count, collect, and split today. I can't put it off until tomorrow. It's collecting every 2 days and splitting every 4.
I have some time until their machine warms up, so I grab the 2 plates I need for photos (FBS Day 6 & ADT Day 6) and head down to the Nikon microscope one floor down. Dr. Chen is finishing up on the microscope and I ask her if I can use her hood to collect and split my cells. Hallelujah, she says yes!
THANK GOODNESS FOR DR. CHEN!
[EDIT JUNE 29TH, 2016 : TAKING OUT THE RESULTS FROM THIS BLOG UNTIL I HAVE MY THESIS ACCEPTED AND PUBLISHED. LAB DIARY WILL BE MOSTLY DOCUMENTING DAILY LABLIFE, PROTOCOLS, + TROUBLESHOOTING FROM HERE ON OUT.]
I text Kyla (our awesome lab tech) to give her a heads up in case she needs anything from the room they're shutting down.
I run back up after photos and grab EVERYTHING I could think of needing--including the very precious cell counter!
I run back down and set up. Literally run-walking around at this point because I don't want to take too much time in someone else's hood, and I also don't know where everything is in someone else's lab.
Anyway, so I successfully count & collect FBS & ADT Day 6. I also split 4 plates --> 6 plates for each set (FBS & ADT), meaning I'm back to having 12 plates again.
|Start with 6 plates ea.--> analyze & collect 2--> 4 left--> split 4 plates ea. back into 6 plates ea. Lab magic, voila!|
NOW, I run back upstairs to put everything away and realize, I need the incubator in the room they're shutting down. Luckily, the guys opened the room back up but were still trying to figure out the hood, so I could store my precious plates. If not, oh heck noooo. I'd beg someone to let me incubate my 12 plates in their incubator until I could transfer them back to ours haha.
TURNS OUT, I regained access to our incubator but our hood will be out of commission until further notice. NO ONE can use our hood AT ALL because of some live wires.
The guys phone called out to see what they could do, but for now...no one can use our hood. Of course, my project is ULTRA TIME SENSITIVE, so...at least, now I know and can plan what to do for the remaining 8 days.
I'll ask Dr. Chen if I can coordinate with her to use her hood again so that I won't impinge on her lab time. She's so awesome, I'm really grateful for her!
Before I call an Uber over to Center City, I figured I'd might as well make protein lysate for the days I have so far of the new collection: Day 0, 2, 4, & 6 (today) of FBS & ADT.
Right now, I lysed Days 0, 2, & 4 of each set in an additional 100 uL RIPA and am spinning them down for an extra 30 mins at 15k. If this doesn't work to get all of the lysate, I'll call it a day and try again tomorrow.
I'll post the full amounts of RIPA and respective pellet sizes later.
I stopped this post after trying to collect the supernatant for Day 0, 2, & 4 again. I'll have to try Days 0 and 4 of the FBS again with more RIPA & more centrifuging tomorrow.
I'm continuing this post from a zen cafe in Center City after my meeting.
For now, I'll leave the cell count data from earlier.
Note that it's currently in concentration--I'll have to calculate out the number of cells from what the data reads.