Please excuse me while I have a breakdown.
I spent over 9 hours alone in the hood on Friday. I came into the lab a little before 6am to work on my protein purification (the latest tally: Protein 4, Julie 0 -- no matter how many/what type of tags I put on this protein, it WILL NOT PURIFY. It outsmarts every purification bead/column/etc. I am working with the two most stubborn, pain-in-the-ass proteins in all of cancer and I am not happy about it), settle down in the hood around 9am, and stay until a little after 6pm. That's right. The whole lab leaves to go out to lunch, and where am I? Oh right, the hood. PI is out of town so everyone else leaves rolls in after 10am and leaves by 4pm and where am I both before and after they come and go? Oh right, the hood. Seriously, by the end of the day, my ass was numb and my eyes were crossed from sitting there for so long. Have I mentioned that I don't even like cell culture to begin with (hence why my project leans towards biochemistry and medicinal chemistry)?
I should also mention that when my PI dumped all this cell culture on me, he said it would take "a couple of hours to complete." In my book, NINE HOURS is not a "couple." Nine hours is ridiculous, when I'm also working on protein purification and crosslinking projects, plus working with a mathematician on campus to generate a formula to help with the screening of our small molecular inhibitors. Nine hours in the hood to make drug dilutions, split drug stock cells, set up 96-well plates for MTT assays, multiple 12-well plates worth of soft agar studies, and take care of my stock flasks, all which require different drug dilutions. And I'm supposed to do this twice a week for the next month, providing the rest of the lab doesn't riot and stab me to death with pipette tips over the fact that I am basically blocking off entire use of one of the hoods two days a week. (In my PI's defense, I don't think he was being an ass and intentionally lying to get me to do all this work, I think he's just so far removed from bench work that he has NO CONCEPT of how long things actually take. This problem happens a lot. Which is why a 12 hour day for me is now the norm, instead of the exception. Remember that goal I had of not working more than two 12 hour days/week? Hahahahaha.)
5:45am this morning I come in to discover that my cells are almost all contaminated. Mind you, minus the fact that I am left handed which is a GIANT hindrance in cell culture hoods in this lab which are set up for right-handed use (I am always crossing my hand over my plates through my "sterile field" to reach for pipettes, the vortex, and so on), I am always very cautious and aware of what I am doing. I've only ever once, in two years of cell culture, had any sort of contamination, and it was from thawing out a new cell line which someone else had frozen, so chances are that the contamination occurred prior to freezing. We appear to be the only lab in this university that tests for mycoplasma on a bi-weekly basis (or ever, for that matter), and I've never ever had mycoplasma positive cells. Never any fungal infection, no bacterial infections of my passaged cell lines. Maybe this is a sign that asking your graduate students to do nine hours worth of cell culture (with the same cell lines, meaning that most of my cells had to be sitting out for a good 6 or so hours) really isn't a good plan. Maybe, by the time your ass is numb and you have a giant sized headache from staring into a microscope counting cells for 2 hours (because he doesn't believe in the accuracy of the coulter counter so I have to count all my cells by hand/eyeball with the good old hemocytometer which only takes 1342 times longer), you start to get a little sloppy despite your best intentions. Maybe, when you're getting in the lab before 6am, you start to get a little anxious to leave by 6pm, especially when you have to head out of town for a wedding. My reward for doing all the cell culture without complaining is that now I get to sit through an inevitable giant lecture from my PI when he comes in later today and starts poking around my incubator to look at my cells, about how I'm not careful enough and not responsible enough and all I'm doing is wasting money, etc etc.
And people wonder why I am perpetually in such a miserable mood. Blame the cells.
Notice that my pipettes are now neatly hanging on the wall. For a year and a half I have been secretly wishing that someone with a pipette stand would quit/graduate/move to another lab so that I could take over the stand. I'm a very tidy individual, and having to throw my pipettes in a drawer just wasn't cutting it (although, mind you, the pipettes were always in the drawer in size order, so I always knew when someone else had been using my things). It turns out these velcro-to-the-wall pipette stands had been in the lab the whole time -- it's amazing what a little bit of cleaning can locate. And please, no making fun of my craptastic Finnpipettes. They are the worst pipettes ever. If you're in charge of pipette purchasing for a lab -- STAY FAR AWAY! I have to calibrate them every single week, and the p2 has to be calibrated every time I use it. They are awful. Now I'm just hoping that someone with the Eppendorf pipettes quits/graduates/moves to another lab so that I can snag myself some actual functional pipettes.
