More experiments

Sometimes I think it will never end. I have just a few experiments left, so I thought. Then I met with my boss today. Like I mentioned before, overall the departmental retreat went very well. My data was well received. One part I didn't focus on, because I was hoping after a few days if I didn't mention anything my boss would forget. Well, he didn't forget.

At the retreat were two of my committee members, Scope Man and Friend of Scope Man. Scope Man had a long talk with Kiwi before Kiwi even gave his presentation because Kiwi whipped out his computer to show Scope Man what we had. And Friend of Scope Man talked to both Kiwi and I after his presentation. The result - lots of suggestions of experiments I should do next. Easy in theory, and I stress in theory. But aren't all experiments that way. Things are never as easy as they sound and appear on paper. What I can't get my boss to understand is that just because other researchers have used a particular technique I will be using this same technique in a way it's never been used before. It might not work. In fact, I think this whole new series of experiments might lead to a dead end, but I'm afraid to think of that.

The other unfortunate thing, in order to run all these new experiments I have to learn how to use a new cytometer. I'm hoping it won't take too long to get the hang of it. I know flow and how collecting and analyzing events works, I know all the ins-and-outs of flow (I was dubbed Flow Goddess by our old lab manager! And the Goddess thing boosts my spirit.). But the software for this cytometer is completely different. Kiwi wanted me to stay with the machine I use now and didn't like the idea of my using a different cytometer. Then I broke the experiment down into smaller pieces and made him understand that for what my committee wanted to learn, I had to use this new cytometer that uses more colors. I had wanted to defend in December but now, I don't think that will happen. I really, really wanted to be done before Christmas. But it looks like I may not be done until January or February. But neither of us mentioned this during our meeting today.

To be honest, I really don't care too much as long as I'm done by April 2008 graduation and as long as I get paid. That last part is the iffy one, finishing by April 2008 is the easy one. I hate talking about funding with Kiwi, it is such a touchy subject. I think I'll give myself to the end of October to see how much I have done (and the likelihood I'll be finished with all experiments by then is slim, the end of November looks more likely), then I'll talk to Kiwi about an actual completion date and setting more limits to my experiments. I hate having to stress about funding when it's supposed to be his responsibility. He never should have hired a postdoc when he can't even care for his own students.

Sometimes I wonder if I will ever get done. I wonder if my committee will ever let me leave, or more importantly, agree to let me leave before my funding is cut.

UPDATE:
Forgot one thing, all those images I made last week, that took the entire week to make, we decided today that they would look better if I did them in a different way. So they all have to be redone. When am I going to do this, on top of all the experiments he just decided that I need to do? Don't ask me. I have no idea.

More building malfunctions

Yesterday at the retreat there were other people who belonged to the Center. I learned something very interesting, kind of worrisome.

The animal facility that is part of the Center tested the shower system Thursday. Apparently, people have to shower when they leave the BSL 3 facility. Well, they filled the shower stalls with water and water leaked into the 7th floor! Holy heck was raised. The lab the water leaked into was the lab of someone important. He said that the Center should be moved to the 7th floor so we would have to deal with the water problem. Everyone has a right to be concerned. This is supposed to be a contained BSL3 lab, and the decontamination shower is leaking. The drainage pipes are double so if the inner pipe leaks there is an outer protective layer. They think a seal under the floor isn't working properly so yesterday they were going to check the showers sequentially to find exactly where the problem is centered. But they will probably have to tear out the floors and redo the seals.

All these problems in this brand new building, that will be working on some serious pathogens has me, and a lot of other people very worried.

We are officially open

Today was the inauguration of the Center and its BSL special lab/primate facility. We've been here over a year but I guess now there is enough equipment and people to make the tours interesting. Last week we were told to straighten up our benches, desks, TC rooms. And they've been telling us everyday to wear lab coats, like we are supposed to do all the time anyway. I spent Friday afternoon making a TC room look neat that we aren't even using yet. Then this morning there were still boards in the hallway without any posters so they were asking everyone who had one to put it up. My most recent poster is in Geeka's office but I had an old one. They didn't care, the people who were walking through wouldn't know any better, they just wanted something to cover the walls. It had been sitting there so long I had to dust it off! I also hung up one of Mo's. They also asked us to empty any full bio hazard bags. One of the techs specifically asked me to gather our stuff up because she was taking it downstairs. She noticed a full bag. I told her that it probably wasn't going to happen. It was LB's trash and no one is going to take out his crap. I just slid it under his desk for her.

Last week they had photographers in taking pictures of people working in the BSL lab, but it wasn't operational yet so they had to carry down supplies to that floor and people pretending to pipette things. Because the animal facility isn't actually ready to open yet. We are just pretending.

The media came through earlier, though I'm not really sure what media. I saw dudes with cameras walking around. Then this afternoon there was a reception where the head of the Center gave a speech, along with other important people. The senior vice chancellor read a letter from the governor - blah, blah, blah.

FD and I purposely didn't bring lunch today because of it. One thing about the Center, when it wants to impress people it pays for good food. My belly is full of good stuff, and some of it was so fancy I'm not even sure what I ate, but it was good! We also got a book about an infectious disease and its eradication, as well as a free laser pointer.

After the reception they started giving tours to people. I don't know who most of them are, actually, I didn't know most of the people at the reception. You could tell who were the students, they were the people in jeans, everyone else had on suits.

At least tomorrow it goes back to normal - disordered TC rooms, benches, desks and traipsing around with no lab coats!

A day of microscopy work, ending with a wet cat

Today was a day full of microscopy work. I spent the first several hours (9am-12pm) in a small dark room, sitting in a chair that was too short for the scope, scanning over several slides that I've looked at before. I had to send some images to Geeka today and I was determined to find what we were looking for. I think I looked at every single cell on those slides. But success!! I sent her the pictures today so hopefully they are what she needs.

I must admit I was also a bit selfish because by spending time scanning her slides I put mine off a couple hours. I was afraid to look at them. I have been working on optimizing this experiment for a year now (at least that long). Normally slide staining and microscopy work is straightforward but my experiment has several variables that make it a bit tricky. Some parts of the staining became complicated, as well as the experiment that was to be stained. Several times I've thought "This is the one. This is the last time I will ever have to do this experiment!" But every time something different needs to be worked out. This time I felt that this really was the last one. I had done preliminary staining a couple weeks ago and everything looked beautiful. The images came out so good that the Kiwi asked if he could put them on his new web page. My hopes were very high this time.

All that was left was to run the experiment then stain as I did during the trial run, just with one extra antibody in the mixture. Unfortunately, it is the pain-in-the-ass antibody. It is an anti-egfp conjugated to AlexaFluor 488. For those of you who don't know antibodies, this one is looking for a green fluorescent protein. But the color of the antibody (the 488) is also green. So how do I know the antibody concentration is right and is actually binding to what it is supposed to be binding to, when it is green antibody binding to a green protein? For various reasons my committee wanted me to do this, mainly because we thought some of the egfp was being degraded within the cell so hopefully using the egfp antibody would help detect more. But who knows? That is just one complicated aspect to an experiment that should be straightforward.

So the moment came. I was done with Geeka's and I put on my first slide. A little less egfp expression than I would have like but that's okay, that has nothing to do with the staining, it was 'one of those other variables.' I had three colors to look at. . . . and only one looked good, that was the egfp. The organelle staining didn't work. It was very faint, if I cranked up the setting on the scope I could make out the organelle but the background was too high for publication purposes. And the membrane marked I used was extremely faint on the conditions where I used the anti-egfp antibody. Which makes NO sense.

Once again I am disappointed. I'm starting to wonder if this will ever work at all. It's stressing me out. My project doesn't have enough substance if this doesn't work. There are just so many variables. When to add the inhibitor, what concentration to make the inhibitor, how long to leave the cells together, it goes on and on. I'm just so frustrated. I told the Kiwi I'm dropping the egfp antibody. If that doesn't work, I don't know what else to do. I also have a lot of flow data to analyze which I've been putting off. It is essential to my dissertation and I just don't want to deal with it if it didn't work either.

The problem is I'm running out of time. I know I could work these out if I had more time to optimize all the variables but I need to finish asap before my funding is cut off. So we are trying to do as much as we can, as fast as we can. And my boss really isn't into "optimizing" things, he just wants to jump into it and doesn't understand why I can't get it to work.

But enough of that, it depresses me. On a brighter note, Scope Man, the head of the imaging facility and committee member, requested some live-cell microscopy movies to show at a meeting he's going to in Moscow. He's not talking about my research but the technology that collected the data. I spent the afternoon trying to get the movies the size that he needed them, less than 20MB. Since he gave me only a few hours notice, and it just happened to be the day I wasn't in lab, I couldn't get them to him before he left. Since his talk is on Thursday he said to email them. Well, no matter what, the smallest I could make the movies was 80MB. So one of his employees suggested I put them on their website and he could download them from their server. So that's what I did, hopefully it will work. It's exciting that he wants to share some of the stuff that took so long to perfect. Even if he does forget to mention whose data it actually is that he's showing!

And the wet cat . . . while I was relaxing in the tub, trying to read a magazine, Dante was staring at me, of course because I wasn't paying attention to him. Sophie wanted to see what he was looking at so she jumped up on the rim and slide right into the water. All the way up to her neck. I had wanted to grab her and hold her next to the tub so Fiance could get a towel but she was able to jump out too fast. It was all Dante could do to get out the way and try to stay dry. There was water everywhere! That small cat soaked up a lot of water. Water all over the bathroom, down the hall, in the bedrooms. We managed to get her and together dried her off. She yowled and growled the entire time. Dante wanted to know what all the noise was about so he kept sticking his face up to hers while we were drying her. She was not thrilled. When we got her mostly dry we gave them both treats. They had a rough day, they were locked in rooms because the air conditioning dude was supposed to come then this happens. And he didn't show, so they have to be shut-in tomorrow again.

An exciting end to just another Tuesday. Life is never boring with cats. That's why I love them. No matter what kind of day I'm having, they always make me smile.

How do you know a student has been gone too long . . .

when the PI has to give him a tour of the lab. Yep, that's right, Kiwi had to give LB a tour of the lab. In the time he's been gone the major changes include the moving of several rooms.

We can no longer use one of our hood rooms, the one designated for a specific infectious disease. The small hood that belonged to my lab was put in the DNA/RNA room while the small hood of the other lab was moved from the general area to where our old hood went.

So we no longer have a DNA/RNA room. Everything in that room is in the aisles of the currently unoccupied benches. Including some of the stuff from the old hood room that doesn't fit in the new smaller room. We still haven't figure out where we are going to put everything. The Center likes to take our space away. Eventually we aren't going to any. Follow all that?

So LB needed a tour. And Kiwi looked happy to do it. While this was going on, FD and I learned what the monkeys feel like at the zoo. In the Pittsburgh zoo the monkeys are behind glass. So people are on the outside looking in at them. FD and I were working in the hood room, with Kiwi and LB talking, smiling, waving their arms, and looking in at us. I felt like I was on display.

It's amazing how someone can show up and immediately put everyone in a bad mood and make the atmosphere change. On a good note, I managed not to actually speak to Kiwi today.

They're baaaack

LB is back. Now I know why. The Kiwi just showed up. He's in his office.

Words can't really express what I feel right now. Hell begins again.

My Internal Dialog

Ambitious Self: I really need to work this Saturday and Sunday.

Procrastinating Self: Yeah, but you've been so tired lately you need to take a day off.

Ambitious Self: But the Kiwi has been gone for two weeks and expects me to have a lot done for our 2 hour meeting next Friday.

Procrastinating Self: Okay, work Saturday but take Sunday off. You need at least one day.

Ambitious Self: But the stupid Center is shutting down the hoods on Monday so I can't do an experiment, so I should work Sunday.

Procrastinating Self: So what, that's not your fault. You can use that day to work on the figures the Kiwi wants you to have done for next Friday.

Ambitious Self: Hmm, you have a good point. I haven't made time for that yet - but I can't work on the figures if I don't have the experiments completed. I could work on them during my incubation periods.

Procrastinating Self: You always say that but it never happens. Your incubation times are too short for you to accomplish anything, and your experiments are too long.

Ambitious Self: But it's a TWO hour meeting. I need to have a lot of stuff to show him.

Procrastinating Self: It doesn't matter how much you have. He won't be happy with it anyway.

Ambitious Self: I really should work on Sunday.

Procrastinating Self: You know you won't. Just face it, stay home, sleep in.

Ambitious Self: But I'll feel guilty for not getting done what I need to get done.

Procrastinating Self: So what.

Ambitious Self: Why should I listen to you? It's your fault I don't have this stuff done already. I haven't done much the past week and a half the Kiwi's been gone. If I hadn't listened to you all this stuff would be done by now and I wouldn't be in this predicament.

Procrastinating Self: You work best under pressure.

Ambitious Self: I really should work Sunday.

Procrastinating Self: We'll see.

Ambitious Self: Yeah, We'll see.

To be continued . . .. . .

I'm back

Well, almost, to the regular schedule thing. I've been putting off doing regular work for awhile. I'm not sick anymore, still coughing because of my pain-in-the-butt asthma but oh well. I can't use illness as a reason not to do work. Lately, it's been easy to come up with reasons not to do work. I know I'm getting burned out but now it's time to go back.

It's not so bad really. The kiwi boss man is gone on a holiday. Won't be back until the second week in August. And LB is on a month's long vacation. On one had, we are all pissed because he can take all the time he wants off. On the other hand, he's not here, which is great. So right now it's not so bad going to work. We don't mind it as much because the two people in the lab (Kiwi and LB) are gone. And no, they aren't on vacation together.

Some updates:

• Ding is back from maternity leave.
• I'm finally going to get my name on a paper, other than a review. I did some microscope work for Mo.
• Had lunch with my class, all the people who came in the same time as me. But that will be another post, lots of issues there.
• Started golf lessons. A reason to get out on the weekend, instead of sleep or work. And I must say, I did awesome!
• My tomato garden is really taking off. Blossoms are starting to show.
• I get to start working on my first author paper. I have to have the layout of my figures worked out for when Kiwi comes back.

Other than that, not too much has been going on. It's been really slow around here.

It's nice to be back blogging! Now I have to catch up on reading my favorites.

Safety measures, yeah right

So I mentioned that we had a Center wide meeting, that lasted 10 minutes, about the tissue culture rooms. I want to reiterate how stupid and pointless this meeting was.

Right now there are two hood rooms, my lab brought over three hoods with two in one room and one in another room. The room with one hood we can no longer use. It has been designated "infectious aerosol virus" room. One of the other labs works with a virus that can infect people via inhalation. A bad virus. So they now have that hood room to themselves.

The safety level is designated BSL2+, which for those of you not familiar, this uses the safety precautions of BSL3 lab in a BSL2 environment. The important thing to note here is that there are NO double doors. There is no double entry. The way it should be for this virus is when they are done working with it, they leave the hood room into an intermediate room, remove their PPE, then leave that room into the outside environment. This way the risk of virus escaping the room is minimal.

Of course, that's ideal. And the brand new facility isn't exactly ready to work with viruses like this. There is no double room. The instructions are that when they are working on this virus they must wear their PPE, including and N95 respirator and post a sign on the outside of the door. When finished, they take off their respirators, leave the room, and set a timer on the door for 30 minutes. It takes 30 minutes for the virus to dissipate from the air and it will be safe to enter the room without a respirator. That is the exact order.

Think about this - remove the respirator then leave the room. What point is there in that. And not permit anyone in the room for 30 min. It's unsafe for 30 minutes, yet they can immediately open the door when they are done to the outside environment. Technically, they should have to stay in that room for 30 min with their respirator on, then take their respirator off, then leave the room.

Oh, the respirators are also supposed to be stored outside the room in the hallway. Because the lab manager said that there should be no infectious virus in the respirator. Wait a minute - am I missing something? Isn't the respirator supposed to block virus from being inhaled? Meaning there is a good likelihood there is virus on the filter?

So, in this 10 minute meeting - we learned the "proper and safe" way to utilize the new room when infectious virus is in use. Boy, I feel safe.

Random updates

As I'm sitting here waiting for my timer to go off, I realize that during the time I was MIA on my blog a few things of note have happened.

• Ding, our lab tech (our ONLY lab tech) had her baby early. Only a couple weeks early so the baby was fine. Our boss was going to visit her but never got around to it. She lives near me so I'm going to deliver her a care package. I wanted to give her some time to settle in with the baby first before I came barging in.


• My boss got a puppy. Just what he needs. Now he leaves early to take the puppy out. He really should be a stay-at-home parent. He doesn't balance family very well with work. Not well at all. His wife wears the pants in the family so he has to work around her schedule. Right now she's gone to a conference or something back in his home country - and left him behind. Hee-hee!


• He hired a post-doc. Our mission to thwart him has failed. Mainly because we never met this one, neither has he. Kiwi doesn't have enough money to bring the post-doc for a visit (he's in another country) so he hired him sight-unseen. Great, just great. He's not coming until the fall so I hope I'm gone by the time he gets here.

I know there's more but beep-beep-beep, I have to go.

Another reason to get done

Geeka will appreciate this. So will Geeka507.

I decided today that I need to get done before I permanently injure myself or break something. Again. I wore a dark purple shirt today. When I got to work only then did I realize that it matched my finger. About the only thing I can do with my finger that doesn't cause pain is type, because the end of it is kind of numb.

I have cut myself with a broken coverslip, cracked my head off an incubator door, gotten an unidentifiable rash on my arm, gotten blisters on my toes, and smashed my finger. And this is just in the past few weeks. You don't want to know what's happened in the past few years. It seems the longer I am a grad student the more that is happening to me.

It is extremely difficult to remove pipettes from rappers, uncap reagents and flasks, and so forth when I can't use my pointer finger. And I have to be careful because I'm working in the hood with blood and everything has to remain sterile.

So now that I'm done with blood I quit. No more today. I have microscopy data to analyze and more experiments to plan but I'm not going to do it. I'm going home. Before something else happens because I can't work properly.

I'm starting to wonder if I'm cursed. Or if there is a grad student curse. The longer a student seems to remain a student I notice the more physical ailments they have. And it's not just me!

Typing with one hand.

So this is the end of a completely unproductive work day. I got up early this morning, as mentioned in the previous post so I could get work done and go home early. Now I get to go home early with no work done.

Earlier today I went to sign up for the flow cytometer. A technician was performing a tune-up with the machine wide-open. I able to see the sign-up sheet but not reach it. I couldn't get on until 6pm. I was going to leave a note on the computer to sign me up when the tech was done but didn't have time because the fire alarm went off and evacuation of the building was ordered.

I went back later to write my name in so the last person wouldn't turn the machine off. The rep was still working on the cytometer. I had baaad feelings. So I went to track down the people who are in charge of the machine to see what was going on. When I went in their office the large, heavy, I repeat heavy, metal door smashed closed on my finger. The end of my pointer finger is now red and purple and almost twice the size it should be. I'm keeping it in ice to try a stop the swelling. It doesn't bend properly. I can't use it very well.

So not only is my finger smashed, my experiment is ruined. All my cells and time have gone to waste. I'm more pissed about the time. I have to run the cells the same day as I run the experiment or they aren't any good. So I've just completely wasted a day.

Maybe I should just forget about my good intentions of getting up early to get work done and get home early. It just backfired.

I'm going home.

An explanation

I feel like I need to explain why I haven't posted in such a long time. It's not because interesting, ridiculous things haven't been happening. Many times I have thought "Damn, this would be a great post" or "I can't wait to talk about this!" But alas, experiments have gotten the best of me lately.

I feel like I have been getting up, going to lab, working, going home, sleeping, then repeating the same cycle over and over again. With some eating here and there. Actually, it didn't just feel that way, it was that way. On the weekends when I finally had some time to relax and spend with family (both cats & people) I couldn't bring myself to turn my computer on. One reason was I can't seem to turn it on without checking my email. I didn't want to check my email because I didn't want to hear from work. The only weekend I actually got on my computer was when I went home to my parents house for my dad's birthday/father's day. I needed to show him somethings on my computer - and dial-up is so slow at home that I didn't bother logging on.

Here is an illustration of how slow the Internet connection is. And it isn't because DSL has spoiled me. They live so far off the main line it is verrrry slow. A program that took me 7 minutes to download at my apartment took over 4 hours. It was sooo frustrating because we got my dad an iPod nano and I was trying to download iTunes. He couldn't use it until I got the software. Then we had to format it for a Windows computer, which it wouldn't even do because it took so long the program would timeout. I had to bring it back with me and it literally took less than a minute to do at work. Now I have to go back this coming weekend to return his iPod and teach him how to use it.

So today I renew my commitment to write. It also helps that I could have actually been home by 6:30pm this evening. But nooooo. I couldn't get on the cytometer until after 6pm. I'll be lucky if I'm home by 8pm.

Reasons why I hate today

• I stabbed my finger with a broken coverslip. FD extracted it but it hurts to do anything. I can't wear a band aid because I can't get the glove over it. It is still bleeding because I'm using it. And throbbing.
• It is now pouring outside. A downpour. Mo jinxed me, she said it was no longer supposed to rain today. Now I have to walk down the street to pick up my blood to process. I told her I should make her do it.
• I put the wrong antibody on my slides. Last night I blocked my slides with a Fab fragment before adding the second primary antibody which is the same species as the first primary antibody. I put on the first secondary again. So I just washed it and added the second primary without blocking again. It better work.
• I don't have time to analyze my data before my lab meeting tomorrow. Because I messed up the slide procedure it took twice as long as it should have.
• It's freezing in here. We now have to wear lab coats all the time but I'm wearing a sweater instead, the coat is too thin.
• I'm starving. I don't know if it's because of the long hours I've been working but I've been ravenous these past couple days.
• The air conditioner is confirmed broken at home. And since we have to close the windows because of the rain it is going to be very uncomfortable. Once I finally make it home.
• I have a sore spot on my head from where it collided with the incubator door yesterday, rather hard I might add.
• I'm tired.

Update:

• This is the punishment I get for multi-tasking. I had three flow assays for today. One completely stained on Tuesday. One fixed yesterday and stained today. One fixed and stained today. While I was transferring the cells from the 96-well plates to the FACS tubes I put one of my positives controls in the tube of another experiment's controls. I don't know if I'll be able to salvage them. At least I didn't screw up the really big experiment from today. Yet.
• At this rate maybe I'll be home by 1am only to be back here at 7:30am to finish processing blood before the necropsy samples arrive. Maybe I can talk my boss into moving our 3pm meeting to Monday so I can go home and sleep.
• I'm still tired.

Might be an experiment free week

So I'm still pissed at LB, he sent me an email saying several things, among which I am now "discourteous" and ended the email with 'have a pleasant weekend.' Not to mince words, I wanted to rip his passive aggressive, arrogant, stupid face off. I was once accused of being passive aggressive, so he's also a hypocrite. And as anyone who knows me can tell you, I can be aggressive (or I like to say assertive) and definitely I'm never passive.

Anyway, there is a necrospy tomorrow. Well, I guess today because his monkey is too sick to last until Friday. It's his own fault his monkey's are dying so fast. I was tempted to run 5 minutes late again, this time on purpose just to piss him off. Instead I decided why should I get up early to get done before the samples get there. I no longer do experiments on Fridays that have necropsy's. Why should Mondays be any different? Should I feel guilty using the necropsy as an excuse not to work? I don't think so. And to be perfectly honest, I didn't do the proper prep work for Monday so I'd have to get there waaay too early to be done by 11am.

On the downside, my entire week is pretty much shot before it gets started.

Monday: no experiments, analyze data, using necropsy as an excuse.

Tuesday: no experiments, have to attend a class on how to write an electronic dissertation. And set up for Wednesday.

Wednesday: process blood, maybe do experiment I set up the day before. Haven't decided if I should go to the newly scheduled department meeting. If I do, my day is shot.

Thursday: probably no experiments this day either, promised a master's student I would attend her exam practice because she didn't pass it the first time. Normally I skip these things but Kdog suggested she talk to me and she actually asked me personally instead of just sending me an email. I feel obligated to help.

Friday: blood in morning, and yet another necropsy, designated data analysis day, if I have any. Maybe lit review, which I haven't done in a while.

And just a short statement to clarify why I can't do experiments when anything else is going on - all my experiments have 1 hour timepoints. That is the longest incubation time in an experiment that can last anywhere from 10 to 14 hours. So if I attend anything, a seminar, data club, journal club, anything, I can't do any assays on that day. Kind of sucks.

In all actuallity, I do have a lot of data to analyze right now as well as a lab notebook to update. I have several live-cell microscopy experiments to analyze and they take hours upon hours. Each experiment has about 2o stage positions, and the scope takes a picture of every stage position every 4 minutes for 5 to 10 hours. It takes a loooong time analyze and I've been putting it off. Guess this would be a good week to get it done.

Other good thing, I have to analyze the data in another lab so I won't see the Kiwi.

Money Issues

It's kind of obvious that our boss is not well liked. He doesn't pay for two of his students, the remaining two (Mo & I), he says he doesn't have enough money to keep. Yet he is actively interviewing postdocs and will eventually hire one. So we know he's lying for the most part. As a result, we (FD, Mo, and I) don't care at all how much we spend. We buy what we need and sometimes we buy things we don't need (usually me). Our motto is "Spend the f*rs money. Who cares."

Well, yesterday, our boss has finally taken notice to what we've been doing. As a result of moving to the new facility, most of our purchases have to be signed by him. We used to use P-card, now we can't. And he never paid attention to finances. He came to each of us and asked that we watch what we spend and try to be frugal. Apparently, we all managed to spend 23,000 dollars last month! Most of his money has been wasted on LB's study so none of us really care. And honestly, most of what we bought last month were legit purchases. Mo and I are growing A LOT of dendritic cells, which require IL-4, GMCSF, and a lot of media with expensive FBS. That's probably where the bulk of the money is going. Oh well, if he wants our experiments to work, to even be done at all, we need the stuff.

I'd be interested to see how much the spending goes down after Mo and I leave.

Lab crap

Someone needs to shake me. I'm trying to resist the urge to complete reorganize (or organize) this lab by myself. I can't take it. There is no reason why anything is located where it is. When we lost our storage room last week everything was put above our benches, nowhere near the hood rooms. LB was pissed because he was the one who had to take everything out of the storage room. So he put stuff in no conceivable order. Pipettes are everywhere, conicals are everywhere. Every time I need something it's like a scavenger hunt. The hood rooms are just as bad. He randomly put stuff in those rooms, instead of putting useful stuff in there. I've been fighting the urge for days and I don't know how long I can hold it off.

The area where our lab is located is about 75% empty, but it is slowly filling up. And in order to entice certain scientist to join the place, the director wants it to look as open and empty as possible (shows room for expansion). However, that means that every time someone moves in, we are forced to compress stuff, like losing the storage room. We all had to move sinks down. It's ridiculous because as soon as an area is set up we have to move it. Anyway, our benches were at the end, now there other people are at the end. We all keep walking down the wrong row. At first I thought he was kidding, but he mentioned it again today to LB, who of course thought it was a great idea. He suggested putting nationality flags at the end of the benches to help people walk down the right ones. He's serious. For one thing, the outside aisle barely fits two people, a flag will get in the way, and we will probably hit our heads on it. He can pay workman's comp for someone getting stabbed in the eye. Plus, we just need to pay attention. If we walk down the wrong aisle, it's our own fault. Flags are stupid.

Friday frustration

I was actually going to be able to go home at a normal hour today, i.e. home by 6pm, but alas, the lab gnomes have foiled all my plans. The Central Blood Back screwed up again today. They didn't draw my buffy coat. They said if I really wanted one they could draw one this afternoon. I said it's not that I really want it, I really need it. I have recently discovered that the main reason my stuff hasn't been working for quite a while is that I need fresh dendritic cells. My frozen DCs just don't cut it. This is a wonderful discovery because now stuff is working. The downside is that now I have to process blood twice a week, which wouldn't be so bad except I have to do large experiments at the same time. I can't lose 2 days per week of experiments.

That being said, since I've been sick this week my energy has been consumed quickly so I was only going to process blood today and go home. Now the blood won't show up until around 4pm if I'm lucky. Which means I will be here until 9pm, at the earliest. I could have slept in today. But now I'm stuck here because it is pointless to take the bus home because I'd have to be back by 4pm.

To top it off, my boss apparently isn't here today. I could have left early with no hassle. One bright light, sort of, is that I don't have to come in this weekend. (Except to feed my cells but that doesn't count.) The laser is out on the microscope I use and won't be fixed until sometime next week. Downside - can't analyze my slides. Upside - don't have to come in!

Later in the day:

More frustration, so when I get back to lab from the CBI, I find out that a monkey is "being sacked as we speak" using my boss's words. It was supposed to go down next Friday. This is the second time this has happened and it's getting pretty old. I called my boss because he wanted to talk to FD or LB, neither of whom was in the lab. He was taken off guard because he didn't want to talk to me, but besides Mo I was the only other one here at the time. My boss told me that samples were coming from the facility and FD needed to talk to the vet tech to arrange a pickup here. He said if I see FD or LB to tell them whatever. I said flat out, "Well, LB doesn't come in when he isn't signed up for the hood. He's not signed up today so he probably won't be here. He hardly ever comes in on Fridays." My boss just kind of stuttered on the other end and changed the subject.

Then we went from frantically trying to get stuff done before the samples arrived to slowing down. They were putting the monkey down but snap freezing the samples so we weren't getting anything. Apparently, FD talked to the boss and told him he was tired and already had plans and wasn't going to process any tissue. And LB couldn't be found, once again he wasn't answering his phone. Surprise, surprise. The boss wanted FD to call LB and figure out what monkey is going to be put down for next week (it needs to be decided by Monday), and FD told the boss if he left a message on LB's phone then there was no reason for FD to do it. I guess the boss was pretty irritated. Good for him. It's not our problem that his golden child is never here and won't answer his phone. Why should we do his work when he's off probably smoking dope and being a bum?

Waiting . . .

I'm currently waiting for a member of my committee to set-up an experiment. He's a 1/2 hour late which is a problem because I have to set my part up before he does his part. This means that the longer he takes, the worse it will look because the cells have been in the chamber too long before starting the experiment.

I am at his mercy. It is a several hundred thousand dollar scope that is his livelihood. No one can use it but him. So I'm sitting here waiting, while my cells get unhappy.

Missing the weekend

I had a wonderful weekend at home. I didn't realize how much I missed everyone and everything until I was there. I've decided I need to go home at least once a month to "depressurize." It did me good because I'm even handling the craziness at work better, and things have not gone well over all this week. And it's only Tuesday!

I live in the countryside, a wonderful rural area off the main highway down a dirt road. It's so peaceful there. It reminded me how much I miss living in the country, and that I'm not a city girl. The sun was shining, the birds were chirping, I could hear crickets outside my window at night. It was warm enough I could leave it open all night long. My drive there takes me through Amish country, right where I live. The Amish were plowing their fields with the horse hitches. It was a wonderful site. The drive home was even nice, I was driving directly into the setting sun and the sky was just ablaze with hues of red, yellow, and orange. It made me wish I had a camera to take a picture. It was so perfect.

We had a surprise birthday party for my sister over the weekend so I got to see most of my family on my mom's side. I miss them all.

However, now I'm back at work. Yesterday I talked LB into helping me with washing and autoclaving, which he then proceeded to start then NOT finish. I had a 20 min break to eat and while I stepped out, he left. He left the stuff in the autoclave room, not autoclaved. That wouldn't have been so bad but he didn't even bother to leave me a note. I didn't know he left and thought everything was getting done. I ended up doing it all this morning - and some of it twice! Some of the packets that were stacked didn't dry and were sopping wet. I had to do it over. I confronted him about leaving yesterday and he said he had to go home and write. He's working on a paper for someone in another lab and when he writes he like to do it at home. Bullshit. He doesn't write, never has and probably never will. Other people do it for him. I hate it when people lie. It's so juvenile and pointless.

I also got up early this morning so I could have blood processed before the faculty candidate seminar at noon. I overlayed the blood over histopaque then proceeded to knock all the 50 ml conicals on the floor, leaving myself with a nice homogeneous mixture of histopaque and blood. The extra setup and spin added an hour to the process. With everything I had planned for this afternoon I am still trying to finish it. I'm in another lab right now waiting to set up another experiment with one of my committee members.

One good thing though - last night I had an experiment finally work. Not much has worked in a long time. The bad part is that now kiwi has a lot of ideas based on what worked yesterday. In the span of a few hours he looked up literature, called another lab, and set-up a meeting with someone to collaborate with, without talking to me first. I need to make sure what he wants to do will even work. The right epitope may not be present on my cells. Not to mention we never optimized the T cell assay, I worked on it for over a year with another cell line and never got it to work, now he wants to try it again. I'm giving him until Friday and at our meeting I have to remind him that he says he can't fund me anymore, that the department will not pay for me indefinitely, and adding all these experiments isn't possible. Because what I did worked so well he wants me to use it as my model instead of the other cell I have - this means redoing every single experiment I have done up to this point. He's insane. He acts before he thinks. And he doesn't think very well.