How many new posts constitutes a habit?

I'm trying to get back in the blogging saddle and write more frequently, since my hellish first year is all but over. I won't promise to write every single day - a minimum of twice a week sounds more reasonable. Writing is still cathartic for me, even though I am finally getting a healthy daily dose of social interaction with the other grad students. I'm starting to feel less awkward and clueless all the time, but I still think that I cling to my seat a little too much because I'm afraid of my lab mates being aware of my presence. That thought/sentence hardly makes any sense, but I can't really clarify it. I guess the social phobia is still hanging on for dear life, despite my shrink throwing some Lexapro at me two months before turning me over to yet another new resident. That reminds me - I need to set up a fucking appointment with the newbie shrink before my Adderall runs out. I don't like having to be seen at all, but I have to play by health insurance rules if I want to remain somewhat functional. The initial appointment is the worst. All I want to have to say is, "I'm depressed, afraid of people, and I can't fucking concentrate. Give me some good drugs and I'll get out of your hair". Instead, they ask me to rehash my entire psychiatric history, even though I can see that they've already printed out the relevant information from my online chart. Blah.

I was gonna write about what I did in lab today, but now I'm too tired and cranky. I guess I lied when I said this was still cathartic. Better luck tomorrow, maybe. On Friday the 13th, no less.

And we're off to the races

I'm firmly establishing my own well-worn ass-groove in the leather chair that goes with my desk in the cholera lab. One thing that annoys me about my desk set-up is that the chair is fairly wide and it has armrests. The armrests are so wide that every time I scoot in toward my desk, one armrest or the other hits the side of the desk. Every. Single. Time. I've got half a mind to take a damn screwdriver to school and remove the fucking armrests. If it's annoying me now, imagine how I'll feel after five years of it. Maybe I'll wait until the PI has agreed to accept me as a full-time lab member and gives me a key to the lab before I start disassembling his furniture. Since I didn't put in as many hours during my rotation as I should have (to avoid failing my classes, not making it to prelims, and subsequently being kicked out of the program), the PI is letting me work in the lab over the summer on a "trial basis" to make sure that I will actually show up to work and generate some decent data. So I'm technically not a full member of the lab. However, I have no intention of being kicked out, asked to leave, or whatever. In my mind, I'm a permanent lab member - it's just not official yet. Unfortunately, I'm reminded of my provisional status whenever the PI sends an email to the lab, but only carbon copies me on it. I don't really understand why that arbitrary distinction has to be made in stupid two-sentence emails stating that we're having lab meeting this week at 2:00 pm - like we do just about every week. That bothers me. Alright, that's enough of the complaint department - I'm starting to feel really pissed off about my stupid provisional status, which will get me nowhere fast.

I had a good productive day in the lab today. It almost felt like it did when I worked in Jim's lab at Eastern, when the science determined what time I went home for the day- not sheer boredom. My PI is off this week, so I don't have to worry about him keeping a surreptitious eye on me. Yay! I feel like I made a butt load of media, but I don't think I actually did. Let's see, I made a liter of 5X M9 salts, 100 ml of 10X PBS, a couple of 90 ml dilution blanks, 100 ml of 20% (w/v) glucose, 100 ml of 1M MgSO4, and finally a liter of complete M9 minimal medium. The shelves atop my bench are becoming beautifully lined with bottles of stuff. Most of the bottles are identical, each containing exactly 100 ml of whatever potion I whipped up - just like everyone else's benchtop shelves. I'm telling you - I really feel like I've joined a cult. Thankfully, no one is sporting any forehead swastika tattoos and wild eyes or wearing all black outfits with gleaming new white Nike shoes.  All the perfectly filled and arranged bottles make me feel like I'm doing real science. M9 is a royal pain in the ass. I think that alone took up half my day - just autoclaving and cooling to room temperature. Fucking ridiculous. But, I had to make it because that's what I'm growing my WT cholera and ToxT deletion mutant cholera in overnight for tomorrow's zebrafish infections. I'm just trying out a very basic experiment to see if it works and to see how long the whole ordeal takes. I'll have two groups of four fish each in separate beakers of 200 ml sterile saltwater. I'm going to add my overnight cultures: WT (cholera toxin-producing) to group 1 and ToxT mutant (no cholera toxin production) to group 2 and incubate the fish at 28 degrees for 24 hours. Then, I'll euthanize the fish and collect the beaker water. The plan is to concentrate whatever protein is in the 200 ml of fish water by trichloroacetic acid precipitation - which I've never done before, so will probably screw it up. The multitude of proteins precipitated from the fish water will be used in a cholera toxin ELISA, which I will also likely screw up in some way. The idea is to see if cholera toxin is present in the fish water. In previous experiments, when naive fish were placed into water conditioned by other cholera-infected fish, the naive fish died. It was assumed that the naive fish were killed by cholera toxin that had been secreted into the water, but no assays were done to confirm this.

Oh, shit. It's almost 12:30 and I haven't had my nightly Ben and Jerry's Coffee Toffee Heath Bar Crunch ice cream gorge session yet. And I haven't finished the paper I'm supposed to read for my virology discussion tomorrow during lunch. And I'm still Jonesing for more of my second read of "Fifty Shades Darker" before I fall asleep. I just love trashy sex and romance novels (with a smattering of BDSM thrown in, as the case may be) - I am certainly NOT a fine literature critic, but I like what I like and, embarrassing as it may be, that's good enough for me. And Chris :)

I'm supposed to do what now?

I've been working in the cholera lab since the end of June now. When the PI said that he expects me to be independent, he wasn't fucking around! My research project is centered around developing a zebrafish model of cholera colonization and infection, so I'm continuing the failed cloning that I feebly worked on during my rotation. Also, now that I've passed prelims and thus should know what I'm doing (ha!), I've been charged with developing a protocol to test for the presence of cholera toxin in water that held cholera-infected zebrafish. The PI didn't tell me what strains to use (and what medium to grow them in) to infect the fish, if I should use log phase or overnight cultures, if I should do a time course, or anything. It's been surprisingly stressful trying to come up with a reasonable experimental plan. I'm sure the point of the "sink or swim" approach is that you learn better what not to do by falling on your face a few times, but this is pretty brutal for me at the moment. However, I don't have to run any ideas by him before I try something new, unless I need really expensive equipment or reagents (I don't even know what dollar amount he considers "really expensive"), so at least that's kinda nice.

 I have to rely pretty heavily on the other grad students in my lab - as well as the grad student, PI, and tech in the fish lab upstairs - for help. I hate being dependent on other people for anything. I'd rather kill myself trying to do something independently than to make my life easier by asking for help. I'm working on getting over this. Slowly. It's really strange working in a lab with other people. It almost feels like I've joined a cult. Seriously. Everyone shares all the reagents (down to the same tubes of Taq and homemade PCR buffer), and whoever uses the last of something has to make more. Sharing tubes of reagents is a little weird (why not just give everyone their own aliquots?), but replacing what you use up is not weird - I don't want to make it seem like I think it is. That is a very good thing! Everyone is assigned a certain chore like making bottles of autoclaved water, LB, autoclaving tubes, etc. The one group activity is stuffing tip boxes - which really sounded cult-like to me. I just picture us all standing at a bench in a row, stuffing tip boxes like synchronized automatons. Makes me shudder a little. As strange as all of this is to me, everyone has made it clear that I can do things the way I want to - if I want my own aliquots of PCR buffer, I can do that. But, I don't want to break away from the way the rest of the lab does things - at least not this early on. Everyone has been very accommodating and helpful though.

I apologize for such a boring blog post - I'm even boring myself right now. Hopefully I'll be back to my normal smart-mouthed self before long and I promise that I will write accordingly.

Prelims: conquered. Now what?

Everything should be downhill from this point. And I mean downhill as in coasting, not the crash and burn or decrepit old git kind of way. I passed all three of my prelims - bacteriology, immunology, and virology. I'm honestly shocked that I passed all three. I was pretty sure that I'd have to retake the immunology prelim, but I guess my committee members decided that they had better things to do in August than to roast me over the coals again. I am so relieved and happy that I don't have to worry about prelims anymore. I've been officially vetted and shouldn't feel the need to keep proving myself to everyone ad infinitum. The operative word there is "shouldn't", because I probably will anyway.

This afternoon, Chris, Elliot, and I went to the Immuno & Micro department picnic held on Belle Isle. Poor Elliot was the only kid, so I guess I should be grateful that he only drove me a little nuts. Chris got to put names with faces and I got congratulated by faculty and other grad students on passing prelims. I got roped into playing volleyball (!), something I haven't done since 8th grade gym class. I even managed to hit the ball a couple of times, so I considered that a total success and quit before I could fuck up my sterling record. I also managed to get a splinter in my foot and a nice sunburn on my chest and arms. That's a first for me - the sunburn, that is. We didn't get lost getting there or coming back; it was a pretty good time.

I've chosen to do my dissertation research in the cholera lab, but the only catch is that the whole lab is going to the national ASM conference in San Francisco next week. I guess I get to relax a little bit longer and catch up on vibrio literature. My actual start date is tentatively June 25. I'm kind of excited. We'll see how long this lasts...

Coming up for air

It's been two months since I last wrote anything that I actually wanted to write. I slightly modified the blog description and added a cutesy old picture of me at Lake Huron. I'm being a bit cheeky in declaring that I'm a 2nd year already, as I still have to pass prelims, but I'm trying on optimism for once just to see how it feels. I've been mired in papers, papers, and still one more paper (extra credit for Neuro - due Friday) - mostly trying to appease Dr. Dick. My prelims are the first week in June: Bacteriology on the 4th, Immunology on the 6th, and Virology on the 8th. Each is a one-hour oral exam, so I have to sit in the department library trying to field questions lobbed at me from three committee members (a different set of three faculty for each oral exam). Unfortunately, Dr. Dick is one of these committee members. I'm not sure how to study for this, while not neglecting my classes or research. I will have to figure this out - and soon. I'm a little afraid my efforts to study will look something like this:

Cock blocked (or should it be pussy blocked??)

Much has happened since I wrote last - I had a birthday in March (meh), my 10th wedding anniversary (and 20th (!!) dating anniversary) in April, and went to lunch at Mercury Burger Bar with the other grad students last month as well. This place is literally in the shadow of the old Michigan Central Depot in Corktown. MCD is really quite an eerie, imposing, yet hauntingly beautiful sight - the picture doesn't do it justice:

Mercury Burger Bar and Michigan Central Depot

Oddly enough, we went to lunch here two days after some kid climbed into an open window on the first floor and fell straight down 30 or 40 feet to the basement. Having seen it in person, I can absolutely see why it attracts "urban explorers". The place is so massive and it was so opulent in its heyday; my dad was around two years old when it opened in 1913. I really can't help but imagine a time when Detroit was young and held so much promise; this area would've been humming with the activity of so many people passing through this train depot - to visit family and see the big city, or maybe to go off to war.
Anyway, about Mercury Bar: they have good burgers, a very busy lunch hour (and correspondingly slow service), and the best garlic french fries I have ever had. I have dreamt about these fries, they were so good.
Enough for now. More in less than two months. Maybe.

Not dead...yet

The final rotational frontier.

It's been a while since I last posted anything and, as usual, I feel guilty about this (I knew those years of Catholic school would come in handy someday!). I just finished my rotation in the virology lab. I really learned a lot in the last few weeks: how to run SDS-PAGE, how to do a Western blot, how to do co-immunoprecipitation, beta galactosidase assays, luciferase assays...and how to screw up most of the aforementioned items. I met Henrietta Lacks for the first time last Wednesday. It was a little eerie thinking about the history of her cells as I transfected construct after construct into them. I only made her acquaintance for a short time though - on Friday I told HeLa goodbye and thank you. It was a slightly religious experience for me.

Onwards and upwards! I technically started my third and final rotation in the cholera lab yesterday, although I didn't do anything then. Or today either. In fact, I'm not entirely sure what I should be doing in the immediate future. Something about cloning GFP into something and then getting it into zebrafish?? I will be working on developing a zebrafish model of colonization/disease. This is a newish (funded!) project, which actually sounds pretty exciting. I'm fairly sure that I'll be working with the tech in my first rotation lab to figure out the best way to get the cholera bugs into the zebrafish. The GFP construct is to make it easier to track the specific location of the bacteria within the gut of the fish. I think. We'll see how this goes.

In other news, I did reasonably well on my first immunology and virology tests, and I'm trying to get caught up with reading and writing for bacterial pathogenesis. I have to present a journal club-type paper to the department at the end of the month and prelims are looming large in the not-so-distant future (first week of June). The thought of prelims really terrifies me. Especially the thought of Dr. Dick lobbing questions about secretion systems and quorum sensing - assignment #2 - at me. I'm not getting paid nearly enough for the kind of anxiety I'm experiencing. That's no joke - I think UM and MSU students get several thousand dollars more per year than I do. I feel like I should get a bonus for managing not to get mugged, carjacked, or shot at in Detroit. That should seriously count for something.

Death of a car & rebirth of a car payment

How sad is it that I feel guilty for not having had time to write anything in several weeks? I'm just piling that on top of my landfill of anxiety - you know, because landfills are tall and full of garbage.

On Thursday, January 26th, my 12-year-old Piece of Shit (POS) car died and left me stranded on I-94, making me late getting to the bar (obviously, the last straw for me). Chris and Elliot had to come rescue me and if I'd had a baseball bat handy, I totally would've gone Office Space on it before leaving it at the side of the road. It was the second time in two days it had left me stranded because it overheated for no obvious (translation: inexpensive) reason. We came back to get the car on Friday, hoping that I could get it home without having to have it towed. We didn't have to have it towed, but it took two days for me to get it back home. Saturday morning, I parked it in front of the house in the spot where our old VWs apparently go to die and then we drove to Perrysburg, OH to look at a used car. I drove back that night, in hock up to my eyeballs, in this:

Stock photo from the internet because I'm lame and didn't take proper pictures.

Isn't it just like me to only take a picture of the ass-end of my new car?! Still pretty awesome though!

I bought a 2007 VW GTI Fahrenheit with less than 25K miles on it! It's beauteous. I think it sends one of two possible messages: I've hit my midlife crisis early or I'm clearly not having more children. Or both. The funny thing is that I bought it from the same dealer I bought my 2000 Jetta from, brand new. VW only made 1200 Fahrenheit edition GTIs in the US and 150 in Canada, and they are all numbered (there's even a registry). As you can see, it's BRIGHT orange. It's an automatic, but it has a Direct-Shift Gearbox (DSG) with paddle shifters. Essentially, it has a dual clutch that allows for almost seamless automatic shifting as well as "manual shifting" using up/downshift paddles located on the steering wheel. I've only tried the manual mode once in an empty parking lot, as I'm a little afraid that I will break my new car. It's fast and even the exhaust burble is sweet. The downside to having a fast orange car is that it's ORANGE!!!! I've been driving it like a grandma because I'm afraid of getting pulled over. I've only ever been pulled over once and I've never had a ticket, and I don't want to start with all that now. Aside from salt trucks and road patching crews, I'm pretty much the only bright orange vehicle on the road. If the registry is reasonably accurate (I haven't updated it yet), my #722 is joining only 8 other Fahrenheits in Michigan. As far as I can tell, mine is the only one in Metro Detroit/SE Michigan, so if you see one driving around or cutting you off in traffic, it's probably me. Sorry in advance.