Controlling my nerves

I don't think I've ever been this nervous in my life. And I'm not defending until tomorrow. My heart has been racing and my hands are shaking. I know I shouldn't be this nervous. But I can't seem to control it.

Showing my boss my presentation

Kiwi was out of the country all last week at a conference. Today was his first day back and we met to go over my presentation for my defense Thursday. The reality of the whole thing started to get to me today. I was actually sick to my stomach this morning. No matter what everyone around here tells me, I know it isn't a "formality" with my committee. They have been so unpredictable thus far I'm not sure what's going to happen. I feel like I'm starting to accept the fact that I might not pass.

When we sat down today, the first thing I asked him was if he looked at my dissertation at all. Nope, of course not - but to be honest I wasn't that surprised. He gave his usual list of excuses, but this time he actually said he wanted to finish working on LB's manuscript to have it submitted by the end of the week. That really pissed me off. What about my manuscript! I'm the one with no publications, I'm the one with the dissertation defense coming up with a committee that's demanding a publication, I'm the one who's no longer working in the lab. He's such an ass. It just shows how low I am on his list of job priorities. Well, he's going to have to explain to my committee why it hasn't been submitted yet. He's had it for a couple months now and we told the committee it would be submitted by the end of April or early May. It's out of my hands.

Anyway, I digress. I went through the presentation. He actually had some useful comments that I'll incorporate into the presentation. He felt I needed more background information. I already have 47 slides and now I'll be adding several more. After this advice, he proceeds to tell me my presentation will have a lot of background but is weak/light on data so I should add some things that aren't in the dissertation write-up. Then he left the meeting say how good the presentation was looking and it should be good. Talk about mixed messages.

So will he get a chance to read the dissertation before my defense? Probably not. He told me he probably already knows most of what's in it. (This after he saw data in the presentation, was surprised, and said how good that would be to add to the manuscript that he's never looked at.) I reminded him I made changes based on early suggestions, and he's never actually looked at an entire chapter. So I asked him to look over that one, short chapter and the overall discussion and let me know if there is anything I shouldn't have said, or misstated, or anything that might come up that I should be prepared for my committee to target. He said OK. That means no.

I can't wait to get this done. Whether I pass or not, it will be over and I can move on with my life.

Those several other things

In the last post I mentioned there were several other things I have to do along with preparing my defense presentation. They have to do with the other job I took. (in addition for looking for a permanent job after that!). I currently have a new partial GSR in a different Center. It is a place I'm extremely interested in for a future job. I met with the Director to show him my work and what kind of things I was interested in, and to talk about a postdoc position they might have in the fall. He's also a person in a position to hear about jobs in the field I'm interested in, which I probably wouldn't find out about on my own. Well, in the course of the meeting he made the comment that it was too bad I was on another GSR because he could really use some help on an Institute he was working on. I told him that actually, my funding had ended in my PI's lab at the end of May so I actually was available. He hired me on the spot. And I love it there!! The people are wonderful and he's great. I actually was describing the position to my Mom and became choked up. I know the situation I've been in for the past several years has been horrible, but on the other hand I kind of got used to it. Now that I'm being treated like an actual human being it really shines the light on how horrible the past several years have been. I can't even describe how this new job makes me feel.

But along with it came more work, hence the other things I have to work on. It kind of increased in magnitude over the past couple weeks. First I was helping with the curriculum. Then I was helping with the lectures, then he wanted me to teach a lecture. Now he has decided that the entire Institute class will be web cast and recorded. So I have several things I need to finish for next week before class starts and my lecture on Wednesday. Teaching for him and working for him is a big deal and I don't want to mess it up. He knows a lot of people and is an amazing person to have on my resume as a recommendation, especially in the field I want to go into.

I just need to balance my time. I'm probably going to focus completely on the Institute until after Wednesday then shift over to my dissertation. It's too bad I'm only working for him until the end of July. I really love it there. It makes me want to not settle for just anything now that I'm going to be done, but get something that I enjoy, even though right now all I need is a job, any job to bring in money.

Turned in at last

I just turned in my written dissertation to all my committee members. Boy, does it feel good. And not that I'm complaining, but Kiwi never even looked at it. I gave him another copy Monday after I received a snarky comment from him about "no feedback for two weeks." Excuse me?! I do believe you were supposed to be giving me feedback! He asked me if I thought it was okay. Of course I think it is okay, well, at least good enough. He actually asked me if I had any questions for him or if I needed anything from him. Gee, it's a little late for that considering I had to give it to my committee members within the next 2 hours. I told him if he could at least read the entire thing before my defense and let me know if there is anything he thinks I should be prepared to defend or even modify that I would appreciate it.

At least it's turned it!! Now I can focus on the presentation (among several other things).

I know I've said this before

But really this time, back to blogging. I lost the spirit for a while. So many frustrations that I just didn't want to talk about it. But I feel that it's time again. I miss my blogging friends!

The changes a couple months can make. First some updates. Kiwi decided if I needed more time that I could (should) turn my dissertation write-up to my committee one week before the defense, even though they were required to have two weeks. Needless to say, it didn't happen and due to a committee member complaining he didn't have two weeks Kiwi gave in. Really, there wasn't much he could say. The rest of the committee members didn't care. They could care in the least. Kiwi wasn't happy but oh well, he isn't paying me.

So how does it stand right now? Essentially, my dissertation write-up is finished, and has been for at least a month. I gave it to Kiwi a few weeks ago but he hasn't had time to look at it OR my manuscript. In fact, he doesn't have time for my defense until the end of July. He can't manage to fit 2 hrs into his oh-so-busy schedule. He sucks. It was just a bunch of stupid excuses. Everything on his schedule is waayyy more important than my defense.

However, all is not bad. I managed to convince the appropriate people from my department that since my boss doesn't have time for me until the end of July, which is not my fault, that they need to cover my health insurance until then. They agreed. The good thing about this is that in order to qualify for health insurance they need to give me 1/4 GSR. So I get a small paycheck. Very small, but it's something. In addition, in a position which I'm going to dedicate an entire post, I have a new GSR in a different department. So I'm getting health insurance and 1/4 paycheck from my degree department and the rest of a GSR paycheck from another. Since Kiwi and the Center won't pay me, I just found someone who would. And it's something I enjoy.

So right now, everything is okay, not great, but okay. I'm just looking for job, and preparing myself for my defense, on July 31st!! Oh, and I haven't seen the Kiwi for weeks, just email correspondence. And that's not for lack of trying, he just ignores every single email in which I ask for a meeting.

I called my boss irrational

After much deliberation (which including talking to mom, Geeka, and fiance) I finally faced the reality that there was no way in heck I was going to have my dissertation written by Monday. I felt much better. However, I still had to talk to my boss. The first thing I did was call the person in the department in charge of the students. He was completely supportive, said it was no big deal. It happens more often than not. Don't rush it if I need more time to write. If anyone gives me problems, have them talk to him. I shot an email out to Kiwi telling him I had to meet with him Friday and we decided 8:45 before his day started. He had a meeting at 9:30. I spent all evening and morning thinking about what I was going to say and how I was going to say it. After interacting with his craziness for 6 years I thought I knew how he was going to react. He'd be pissed, complain and whine, insult me, but ultimately say it was my decision and it's between me and my committee, he wouldn't want a part in it.

I thought wrong. He is much crazier than I thought he was - and that's saying a lot.

This was the rational that I would use:

1 - I have been working non-stop but the reality is that 1 1/2 months is not enough time to write a dissertation from scratch. Without a manuscript to start from.
2 - I understand I will not get a stipend after May. He won't pay me. I'll deal with it because it's my decision.
3 - The committee won't care, all they want is a manuscript which had to at least be submitted by the time I defend, which it isn't. It's been on Kiwi's desk for weeks.
4 - It serves no one to turn in something that isn't done; it is a waste of every one's time.
5 - It will not affect him at all since he won't be paying me. I just have to write and schedule a meeting.

It's hard to even describe how the meeting went. He asked how it was going. I said not that great. And told him I finally faced the reality that it wouldn't be done by Monday. We argued for the next 20 minutes. Through the course of the argument I reiterated several times what I mentioned above. Here are some of his comments.

"I will not even entertain the idea."

"I recommend you don't pursue it."

"As far as I'm concerned your dissertation is on the 19th."

And the best one "I'll be there on the 19th for your defense whether you are there or not."

He had the look of a petulant child throwing a tantrum, like he's going to do it whether I like it or not.

Then he proceeded to compare me to a student who had rescheduled several times and was there 9-10 years. I just interrupted him, told him that isn't even a fair comparison. The difference is my work is done, all I'm doing is writing, and I'm pushing to leave. She wasn't.

I must say it was wonderful to call him irrational to his face, and ridiculous. I'm just glad I've developed self-control over the past few years with him. There was A LOT worse running through my head.

Finally, he said turn it in next Friday instead of Monday - which I should be able to do. But I never actually verbally agreed to it. Then I asked him why in the last email he sent me he said the manuscript had to be completely reworked. We went through it, he's only scanned it so far. I asked him if he looked at the part of the dissertation discussion I sent him a couple days ago. He hadn't. I made him open it and go over it with me. I made him 1/2 hour late for his meeting. I don't care. If he was going to be such an ass, he was going to give me CONSTRUCTIVE feedback on what I sent him.

Grrrrr.... I went and vented to Geeka. We calm each other and prevent the other from harming people.

Of all the scenarios I ran through my head that was not one of them. And at the end of the meeting he had to sign the GSR letter stating I will get paid the month of May. He looked at it - "What's this," "What does it say?" and proceeded to read it. It is the same letter he has signed for every student since before I was there - every semester. Like I was hiding something in it!

He's gone next Monday, Tuesday, and Wednesday so I scheduled a meeting with him Thursday morning.

I can't wait to get the hell out of here. Oh - I told him that too. If he thought I was making the decision lightly about rescheduling I wasn't, I want to be out of here and get this done more than anyone. I want to move on with my life.

Dissertation RBOC

I am officially at the I-don't-think-I-can-do-this stage.

I still have 50 pages to write minimum, and finalize this by the end of the weekend.

My boss is being an ass, decided he wants to look at the dissertation today, but I'm not done. So let's see if I can write those 50 pages by tomorrow morning.

The manuscript won't be done by the time I defend, all of a sudden the entire thing needs to be reworked and rewritten. And Kiwi doesn't have time to do it before then. We told my committee it would be submitted. I just think he doesn't want to take time to look at it now.

I still haven't looked for a job.

I find myself thinking of sleep more and more.

Realized today that the student before me got away with using very large font and double spacing the reference section.

This whole process isn't worth it, there's more to life than this. Feel like I wasted 6 years of my life.

Recipe: Mix a pinch of sleep, some anxiety, and a lot stress. Bake in front of a committee for 2 hours. Get one serving of fried me.

It finally feels real

After 5 meetings with individual committee members, countless emails and phone calls, and lots of paperwork . . . . . . ready?? . . . . . tah-dah!! . . . a defense date!! May 19th, 3pm.

I can finally move on with my life. Woooo-hooooo!!!!
It comes with excitement and a wonderful sense of peace.

Best way to format my dissertation ??

I thought I knew how I was going to put my dissertation together. Not anymore. I had a meeting with Kiwi on Friday and he seems to have changed his mind, but he hasn't convinced me yet that this is the best plan. So over the past 6 years my project has been rather all-over-the-board. Every committee meeting my third aim changed. That wouldn't be so bad but I've spent months and months (years) going in various directions, only to have either hit dead ends, or was stopped by Kiwi before I could finish a series of experiments. Thus, I have a lot of experiments (i.e., series of experiments) that were never finished.

I also have nopublication. Kiwi decided a long time ago he wanted my entire dissertation in one manuscript. I tried to change his mind several times but never succeeded. Now he has put me a hard situation. We are writing this manuscript and are leaving several pieces out as 'data not shown." Here's the tricky part, I want to show the DNS in my dissertation. I also have to divide my dissertation into chapters. The standard is to use publications as chapters, which kind of follow your aims. Well, I will have one manuscript which contains ALL of my aims. I can't just have one chapter. So we decided that we were going to break up the manuscript into three chapters, following my three aims and include everything we left out. Sounded good to me! But he's had a change of heart. Now he wants me to focus on my manuscript because of the committee, insert it as Chapter 1, which will encompass all my aims, add the DNS in the appendix, and "make-up" a fourth aim. Yes, you heard me right. That is why I'm here on a Sunday, instead of working comfortably from my apartment. I'm pouring through 5-6 years worth of lab work, a lot of which was never finished, trying to come up with a 4th aim. I was venting to FD and we both agree this is a colossal waste of my time. Kiwi said he hears me say how I've done a lot of work so why not include it in a fourth aim, which yeah, I have, but he's stopped most of it. Now my 'homework' for the weekend is to see if I can make my experiments that were 'not publishable' and didn't work as 'we wanted' fit coherently into another aim. And he said I'll have to figure out some way to explain 'around the data' during my defense as to why this should be included in the thesis since it isn't publishable.

Grrr.

So, I have three days total to 'make-up' a 4th aim, finish my manuscript figures, write figure legends, and finish my methods and results. My series of meetings with my committee members starts Tuesday afternoon and since my committee doesn't care about "aims" I'm only going to talk about my manuscript. But I need the fourth ready in case they want to know if there is 'anything else' besides the manuscript.

And I still need to figure out what the last component of the manuscript means. The data didn't turn as we expected. Which isn't a bad thing. Except now I have to explain it which I'm having trouble doing, and Kiwi doesn't know either. And since it's the last major part, I just know that's what the committee is going to question. When my dissertation became a 'cell biology' project it started being out of my element. I've never even had a cell biology class!!

Just too much

I haven't written in a long time. Really, the reason is that I'm just a bit too overloaded. When I think about writing my brain practically explodes from the things that are running through my mind. Just sooo much stuff to talk and think about. I'll just have to take it in small pieces. After a short crying spell last night I've calmed down and now I'm trying to take control of a situation that has fallen under the vices of others, namely my committee members.

I came back from Christmas break with optimism about what I had left to do. I had everything planned out, I even processed blood two times in preparation. Then everything changed after an email that Kiwi sent me out of the blue, it came after an email requesting a meeting. Here is the main excerpt:

"I cannot buy any more supplies for your studies, which are now not funded,
and the Center needs to stop funding your salary as they are overextended."

After that, all work essentially stopped. I panicked at first thinking that meant next month (Feb) but I found that "as soon as possible" meant April. But now after the past week, I'll be here through May, defending mid-May. That's it. No more. No options. I'm extremely
upset that I won't be done at the end of April because that means I'm just going to miss the graduation deadline.

There's a lot more to this story, which I probably talk about as I have to take breaks or vent in the next few months. But right now I'm a bit overwhelmed because I have to write an entire dissertation and manuscript in two months because of the demands of my committee.

I have so much running through my brain right now that I'm probably rambling, which is why I haven't posted. But maybe if I start typing again those thoughts will become a bit more ordered.

My laptop has a new soul

It's been so long since I've posted or read any one's blog I feel like I'm in withdrawal. It's funny, even though I don't really know anyone out in my blog world (at least, most of you) I still missed everyone!

My computer has been down for a while. And I'm not comfortable using the lab desktop to blog so I haven't done it lately. And as I mentioned before, I'm not good at writing once I get home. I'm not sure how to qualify what happened to it, it was in a coma then awoke for a few days, until I killed it. Now it has a new soul. Let me explain.

1. Coma (data still there, just not accessible) - motherboard needed to be replaced.
2. Revived, awoke from coma - motherboard replaced so it's a happy computer, for a few days.
3. Dead, I killed it (no longer registered hard drive) - It had the unfortunately luck of being on my lap when I got really pissed at someone. While it was running I slammed it shut, physically damaging the hard drive. I lost everything.
4. Receives a new soul - new hard drive installed, so it has the same body but all new insides.

It took a while for me to get everything reinstalled, and I still don't have everything I had before. At least I can start writing when I want to now. And I have so much to read! I have over 300 on my Reader since I've been gone a month. Ugh. Please be patient.

My laptop went into a coma

The past few weeks my laptop has been doing strange things. At random, it wouldn't detect the network or wireless card, it detected unknown new hardware, the USB drive worked only with a few things, and sometimes the power button had to be pushed two or three times before it would come on. All this happened randomly, sometimes together, sometimes alone, but it was always something.

The IT person here at the Center is fabulous. He actually is very knowledgeable and helpful, unlike most IT people I have experienced. I was hoping he could fix it but when we realized he couldn't (which was a couple weeks ago) we decided it was time for me to call Dell technical support because my laptop is under a full warranty. I could even drop it and they will replace it.

I have never had a problem with Dell support, I always got what I needed handled by the appropriate person. I've been procrastinating because I wasn't looking forward to going through all the steps in the support persons "script" because they "have to." (The quotes are from a support person I spoke to a few years ago.) I needed a block of time to deal with it at work (I wasn't wasting time at home, I'm usually too tired anyway) and that time was today. I checked my email this morning, walked away to set-up an experiment, and when I came back my computer was off. And wouldn't turn back on. Looking on the bright side, I thought "at least I won't be on with tech support for long, there is only so much to try when the computer won't even turn on." I should have known better. I really should have.

Here is an abbreviated version of what occurred.

Tech#1: Hello, this is tech support, may I have your service tag?"
Me: Don't you want the express service code?
Tech#1: No, I need your service tag.
Me: 12A34
Tech#1: Okay, you have a model A, correct?
Me: No, I have a model B.
Tech#1: Do you still own this computer?
Me: Yes, it's sitting right in front of me, that's why I'm calling. (Duh.)
Tech#1: Has it always been yours?
Me: Yes, why?
Tech#1: Well, according to our records, you have a model A, the ownership has been transferred to someone else, and the warranty is expired.
Me: No, it isn't.
Tech#1: What is your service tag again?

I repeat it, he still gets the same thing. We argue for a while, with me becoming progressively pissed, and him getting more rude. Finally . . .

Me: Well, your information is wrong.
Tech#1: Let me talk to my manager.

After several minutes . . . .

Tech#1: You need to talk to customer service.
Me: You're kidding me.
Tech#1: (in a snotty voice) No, you need to talk to customer service and verify that the computer belongs to you.
Me: (fuming) Can you transfer me?

Rude Tech#1 proceeds to transfer me and disconnects me in the process. I then go online using the work computer and get the customer service number. I call customer service which has an automated voice telling me there is a wait due to high call volume. In the meantime, I have to get work done so I put the call on speaker phone, I take my cells out of the fridge, then someone picks up. I flag down Ding to put my stuff back in the fridge for me. I immediately start loudly complaining to customer service (CS) about what Tech#1 told me then apologized for being rude to him, I know it wasn't his fault, but I'm pissed.

CS: Can I have your service tag please.?
Me: 12A34
CS: Can you tell me the model please?
Me: Model B.
CS: Ah, are you sure?
Me: Yes, I'm sure. (Of course I'm sure! It's sitting right here in front of me. If I get one more stupid question I'm going to scream!)
CS: That's not what I'm seeing in your account.

I then go into a tirade about how I'm looking at my Dell account online, on the Dell website, and it's telling me that it's the correct computer, it's mine, and the warranty is good until 2008. How can he possible be looking at something different! He then repeats the service tag, which is wrong. He types it in properly and all of my information comes up correctly. He is extremely amused that all the confusion was due to a phonetic misunderstand. Aaaaaaahhhhhhhhh! Rude, and now Stupid, Tech#1 got the service tag wrong three times. What a colossal waste of my time. CS now transfer me to tech support. Again, I am on hold, so I try to do work, take my cells out of the fridge, only to have someone answer the phone. This time I flag down Apple to put my cells back.

Tech#2: Can I have your express service code?
Me: Everyone else wanted the service tag.
Tech#2: No, I need your express service code please.

I give him the number and all of my account information appears correctly. The express service code is all numbers, unlike the service tag (where a letter caused all the confusion), but that shouldn't have mattered because I speak clearly.

Verdict - I need to get a new motherboard. Since the hard drive doesn't need to be replaced, just the motherboard, my computer is in a temporary coma. Not usable, but all the information in it is not lost, so it's not dead.

I complimented Kiwi

It happened before I knew what was coming out of my mouth. I couldn't stop it. I was getting off the elevator and he was getting on. He had black slacks, a red shirt, and a black and grey jacket. I was so stunned that he was wearing normal clothes I complimented him before I could stop myself. He looked almost as shocked as I was.

When things go flying, you have to laugh

I have decided to nickname the rotation student 'Apple' because I have recently learned she loves caramel apples. She made me and Fiance two. They were really good.

Last Friday I agreed to help Apple process blood. Even though we processed side-by-side two weeks earlier, she wanted to go through the procedure again. Since she is eager to learn, pays attention, and takes initiative (unlike PD) I agreed. Even though we had to start at 7am, meaning I had to get up at 5am, and I'm not a morning person. We had to be done before the samples from the necropsy were to be processed.

I wasn't going to do anything. She was going to do the entire procedure, I would only interrupt if I saw she needed correcting or advice, or if she asked a question. Most of the procedure went very well.

The last part of the procedure involves separating out cells that have incubated with beads using a column and a strong magnet. It is a very strong magnet. Well, when she went to remove the column from the magnet she wasn't prepared for the release. It popped right out of her fingers and flew across the hood. I couldn't help myself, I burst out laughing. It was hilarious. She was mortified and felt horrible because she probably just contaminated all of my cells but I couldn't stop laughing. Probably because I was so tired. I just put my arms around her shoulders and told her it was okay. That she should be glad I like her because if PD had done that I would be pissed. She said what I was thinking - thank God it didn't land in the bleach! I wiped the column with ethanol, since the cells were in the column there was a chance that the cells were still sterile, and had her push the cells out with the plunger. She needs to build her arm muscles because she didn't push hard enough to get all the cells out. While I was counting the cells I made her practice putting the column in and taking it out of the magnet. I told her to be prepared because this week she will have a pop quiz. I'm going to get out a column and make her do it again.

The cells looked a bit haggard today, but I don't think they are contaminated.

The new postdoc

Over the past couple weeks I have been subjected to the new Postdoc (PD). After very little deliberation I have decided that if someone like that can have a Ph.D., give me mine right now. Seriously, I've lost my patience with him and I usually enjoy helping and teaching people. The only two people I've ever had a problem with is LB and Kiwi because they bring out the worst in me. I'm trying to be nice, I really am.

And something completely random - PD has this grin on his face every time he talks. No matter what he says. It annoys me. Don't smile when you're telling me you're going to be late. Don't smile when you're telling me you think something didn't work. It's kinda creepy.

Anyway, PD asked me if I would go through the blood processing with him again. He watched me once, but he wanted me to do what I did with the rotation student, a hands-on go through. We decided to work side-by-side in the 6ft hood so we could work together for awhile, then when the procedure called for the joining of our samples, I would watch him. We planned to start at 8:30 in the morning so I would have time to work in the afternoon and meet with Kiwi at 1:30pm. The day before the scheduled work he looks at me and says he's going out the animal facility with Kiwi and won't be back until sometime after 10am (all with a smile on his face). Great. There goes my entire day. I told Kiwi we needed to reschedule our meeting because PD and I would never be done in time. I made sure Kiwi knew he was messing up my schedule.

The next day they finally showed up around 10:30am. Kiwi came to me and said they were back, PD had to do one thing then he could work with me. It was obvious I was impatient. PD couldn't seem to grasp that as soon as he showed up I wanted to start right away. He was late, which was Kiwi's fault, but I had to track him down twice to tell him it was time to get started. I'm taking an entire day to show him something, he needs to be courteous. He could have waited until we had the procedure started to start his computer and check his email.

I have optimized my procedure for speed because I have other stuff to do. I realize sometimes it's hard for me to slow down because I want to get it done so I told him to let me know if I go too fast. Well, I went extremely slow and he still managed to screw up. Here is a summary of the day:

• I made him throw away four pipettes before I was confident that he was removing them from the wrapper without contaminating the ends.
• It took him 1/2 hour to make a buffer that should take, at the most, 10 minutes to make. He was so dirty that FD tossed the reagents PD used because we weren't confident he didn't contaminate them.
• When he tried to collect the white blood cells from the ficoll gradient with the pipette, he was only sucking up 1 ml at a time. I finally realized he had been trying to pipette with the pipetter turn off.
• Once it was turned on, he proceeded to suck plasma up and through the filter. He didn't clean it properly when I told him exactly what he needed to do. FD graciously cleaned it while PD used my pipetter (okay, actually LB's pipetter; we 'borrowed' it from his bench, it's not like he uses it anyway.)
• He consistently dripped whatever he was pipetting (buffer, plasma, white blood cells, PBS)
• Every time I had to pick up his conical there was fluid on the outside. I couldn't figure out when he touched it so I had no idea if the cells were contaminated.

I understand that it can make people nervous when someone is watching their every move. So I didn't get upset with most of those things, not too upset, and I was patient. However, he has done very similar procedures to this before. He worked with the same cell type we do, just from a different species. Things are very similar. But if I someone asked my opinion I'd say he never worked in a TC hood before. When I started to lose my patience was when I would ask him what the next step was and he couldn't tell me. He says a few things and it would be wrong. He couldn't even tell me what step we were on in the protocol. I had given him a step-by-step written protocol that was right in front of him. He's had it for a few weeks. I was thinking - Don't waste my time, which I'm using to teach you how to process blood, when you can't even follow the protocol.

That's when I lost my patience. He's a postdoc for goodness sakes!! He should know how to follow a protocol which is called "Human Blood Processing for Dummies" (no kidding, I wrote it several years ago.) He should be able to use a pipette

That's is the only and last time I will ever waste a day teaching him anything. I understand there is always an adjustment period when starting in a new place but he should at least have a clue as to what he's doing. This is the person that Kiwi hired when he stopped my funding and said he didn't have enough money to pay for me.

The following Monday morning he looked at me and asked what markers we stain for on the cell type we work with. Here was the conversation:

PD: What markers do you stain for on your cells?

Me: Huh? What do you mean?

PD: I'm comparing immature cells vs mature cells. What do you look at?

Me: (looking a bit incredulous) I'm not telling you.

PD: (smiling, always smiling, looking confused)

Me: We don't give the rotation students answers so I'm not going to give you the answer. Look up what you think you should stain for and why, then I'll tell you if you're right.

PD: (still smiling, still looking confused)

Me: Okay, what do you think you should stain for?

PD: (He lists markers)

Me: Right, that's fine.

Do you know how easy that information is to look up? It's even in a previous paper that came out of our lab. It's basic biology. If he doesn't know, look it up. Show some initiative, he has PhD and should be able to answer his own questions like that. Look things up. I'm not giving him the answer. I have my own stuff to work on, he should look it up, if he can't find it, then come see me. I am not the first stop.

I'm starting to wonder if I'm expecting too much of people. I just want people to do the work they've been hired to do, show some initiative, and don't waste my time if you aren't going to listen to what I tell you when you ask.

Time flies

I've been neglectfull of my blog. I've been a bad blogger. I've actually been working all day with no breaks, i.e., no blogging at work, but I haven't been bringing my laptop home so even though I thought about writing it never actually happened. I didn't realize how long it had been until my dad mentioned it last night. I can't beleive the last entry was last month. It's also been that long since I've read any blogs. I have alot to catch up on!! Starting now.

BSL2+ training

We had mandatory training yesterday for BSL2+ work. Forget the fact that we have been working in BSL2+ facilities for over a year. Apparently now we need training. The most interesting thing about the training - the full time biosafety officer teaching the class passed out candy for those who would answer questions or make comments. We were in the BSL2+ tissue culture room. In a safety class, we were encouraged to eat in the same room we were just told that we weren't allowed to eat in.

Ridiculous.

And during the same meeting we learned that the two table top centrifuges are room temp only. They aren't refrigerated. In a common lab space only an idiot would order centrifuges that don't refrigerate. Two of the three labs here use refrigerated centrifuges, however the lab in which the Center lab manager belongs to doesn't. She obviously doesn't think of every one's needs. Kiwi is working with her and the other lab boss to figure out what they are going to do. Absolutely everything our lab does requires equipment to keep it all cold. Kiwi was eyeing the centrifuge next to my bench. He wanted to take the centrifuge from our lab benches but I said no, bad idea. That's the only refrigerated one by the benches. I told him he can't have it. They will need to fix the problem without stealing the equipment the rest of us use. Especially the equipment I use!

Dante's Update

I want to thank everyone for their well wishes. I really appreciate them!

I would have posted yesterday but wasn't ready to talk about it yet. Don't worry, Dante is home and running around like a crazy cat. But he still has his bad tooth. The vet didn't want to do the surgery.

It was a combination of a couple things. Dante wouldn't calm down in the cage, he was panting and breathing hard all day. They didn't want to anesthetize a cat breathing like that. But that wasn't the main reason. He had an EKG before the surgery to check his heart, and it wasn't good. In fact, it was the worst that it's ever looked. The vet described to me what he thought Dante's heart was doing, simply, it was trying to expel blood from the heart when new blood hadn't yet been pumped into it. He showed me the EKG and the irregularities. He said he hasn't seen this kind of abnormality in a cat before and that it was more than just a murmur. No one there (there is more than one vet) felt comfortable putting Dante under with his heart like that. A tooth wasn't worth him dying on the table. So even though I feel that he would have been fine, they didn't want to take the risk since he was so worked up. In fact, they were so concerned about the EKG they didn't even really care about the tooth anymore.

So what are my options now? Well, not much. Here are my choices.

1) Do nothing.
2) Tell them to go ahead with the surgery no matter what.
3) They can bring in a vet specializing in tooth removal so it can done as fast as possible
4) Go back to see the internist that Dante sees about his heart once a year and get him checked out again.
5) Send him to a feline cardiac specialist to determine what specific heart disease he has because there may be a treatment that could help. And the specialist could determine the best way to handle putting him under.

Of course the best option is number 5. And the vet would recommend number 5, because as he put it, he would hate to have Dante develop problems in the future, only to learn later that they could have been prevented, or at least delayed. But of course, the list is in order of least cost to highest cost. A visit to the cardiologist would start between $500 and $1000 and that just isn't something I can afford right now. So as he put it, I'm stuck between a rock and a hard place. He is going to get an opinion from the cardiologist for me for free. The cardiologist visits his office about once a week and he'll show her the EKG to see what she thinks. Then he'll call me.

When I think about it, cats never used to have teeth removed when there was a problem, and they did just fine. It would just come out when it rotted out. I know that sounds horrible. And the tooth eventually will become painful to him, but it will come out on its own. I don't think getting the tooth pulled is worth losing him. But I also know that I can't avoid anesthetizing him forever. Sooner or later, he will have to be put under.

My last cat died of feline leukemia virus. He lived a long life for a cat with that disease, over 10 years, but at the end he wasn't good at all. It was hard knowing that he could have passed away at anytime, with no warning.

And now I feel that I'm facing a similar situation again. The difference is, Dante's might be treatable. Who knows when his heart will become a serious problem for him. I try not to think about it and enjoy all the time I have with him, but every time I go to the vet for his shots and check-up I'm reminded of his problem. Veterinary medicine has come a long way but sometimes I think it is better not to know. We had barn cats at home all the while I was growing up. They never went to the vet, we never worried about them. And they lived happy lives, some very long lives. Veterinary medicine can help keep a pet healthy and happy for a long time. But there is nothing worse knowing that there is medicine and technology out there that can help - and I can't afford it.

I can't help but think that if I had a real job I'd be able to help him now, and not later when it might be too late.

Wish me and my cat luck

Tomorrow my cat Dante has surgery to remove a bad tooth. For most cats this is a minor procedure, no big deal. But not for him. He has a heart condition labeled "feline intermediate cardiomyopathy," in other words, he has atrial enlargement to the heart resulting in a heart murmur. He is on a pill-a-day which has helped immensely, at his last EKG and blood work he had a barely detectable murmur, the enlargement was down to normal size, and the blood work was normal. However, there is an increased risk of complications from anesthesia because of his heart condition. His normal vet didn't want to do the surgery initially until he talked to Dante's heart vet who said it should be okay, as long as they didn't use certain drugs.

So in all likelihood, he'll be okay. He runs around like a crazy cat so I know he feels fine. But I'm still worried. My mom says he'll be fine and I'm sure he will be but I can't help it. There is always that possibility that something will go wrong. My parents are coming out tonight so they can drive us to the vet tomorrow morning to drop him off, then Fiance and I will pick him up in the evening.

So I know I'm rambling but I'm worried. I just can't imagine my life without him. So wish him (and me!) luck, put us in your prayers and thoughts. I'm sure he'll be fine, but I can't relax until it's over with and he's home tomorrow evening.

This is him laying in his favorite position!

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.

An hour I will never get back

I just spent an excruciating hour listening, well, sort of listening, to LB's data club. Which of course, contained no real data of value. He spent the entire hour showing other peoples' work, with some his own, and explaining how his vaccination study didn't work.

Since when is it appropriate to give a presentation of your data, while sitting cross-legged on a table, calling your boss the fearless leader? Data club is supposed to be casual but people at least try to act professional about what they are presenting.

He took credit for a lot of work he didn't do. In fact, he talked about how this construct and virus he made was what he was most proud of in his project. He didn't have anything to do with it! He couldn't clone if his life depended on it. Someone in another lab and Ding made all of his stuff. Data club is a place were ideas are tossed around, suggestions made, stuff like that. Several people were trying to figure out why he got the results he did and what it all meant. When it can be interpreted because you can't conclude anything from a study where the administered vaccine wasn't titered and the drug therapy was miscalculated. Geeka and her boss kept asking him questions trying to expose how much of an idiot he is. It was amusing at least. He couldn't even answer the question about what kind of cell types had the receptor for the cytokine he was using.

It's amazing how my anxiety physically increases when I hear him talk. I mean, I can feel my pulse go up. And an hour was almost intolerable. I wouldn't have been there except I had to escort the new postdoc and I wasn't going to leave early because Geeka came to keep me company. However, it would have been amusing if both her and I and her boss all left at the same time.

FD has some good suggestions for LB to do for his project. The problem is, it actually involves work and LB doesn't do work. In the past 3-4 weeks he's only been lab 3 times, and not to actually do work.

Kiwi wasn't even there, he had a biosafety meeting to attend. But apparently that pissed of some faculty who were there. It looks really bad when the PI isn't present for his own student's presentation.

Geeka and I said in our next data club we should show the mounds of data we have of stuff that hasn't worked, and title it "Can I get a Ph.D. with this?" A professor I talked to with a great sense of humor said "At last, an honest presentaion title."

Desktop

I've never done the desktop meme and I saw it on Dr. Brazen Hussy's site.

I'm constantly changing my desktop - at least once a week. I read somewhere that frequently changing your desktop picture keeps your mind more alert because you are always looking at something different. I don't know, that might be a bunch of crap, but I love changing the desktop with my moods.

This is a picture of some beautiful bright flowers and a bee. I wanted something bright and summery, because summer is gone and I know fall and winter are around the corner. It reminds of warmth, sun, nature and life. Even though I'm not really missing the warmth right now because it is unusually warm for this time of year, above 80F!

This picture is from the photo of the day section of nationalgeographic.com

Whoever wants - it's your turn.

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.

The week was worth it

This week may have been hard but for me, yesterday made all that work worth it. I met Ding in front of her apartment and we left for work to pick up PostDoc then headed out for the retreat. Registration started at 7:30 and talks started at 8:30 but we decided to leave around 9am. Kiwi was inoculating animals in the morning so he wouldn't be there until maybe 11am. We saw no need to get there early. It was in a building located near downtown so we didn't have far to go.

The retreat consisted of various faculty members giving 1/2 hour presentations with 10 more minutes allotted for questions. Unfortunately, the moderator didn't moderate. All he did was bring up the next person's presentation when the current person was finished. As a result, almost everyone talked too long. Some people talked twice as long as they were supposed to but no one said anything. By lunch time we were over an hour behind schedule.

Lunch was at a famous, fabulous Italian restaurant. The entire retreat was worth it because I got to eat there. I always wanted to but never actually got around to it. I'll have to come back with Fiance sometime. But they only scheduled lunch for 1 1/2 hours. Lunch was a few block away from the building where the presentations were located. So over 50 people had to walk to the restaurant, get seated, and be served a multi-course meal. An Italian meal. I've rarely been to a good Italian restaurant that serves food like that and been completely done in that amount of time. So we were running even later after lunch. However, there was a guest speaker who was supposed to speak after lunch and he never showed up. Since he wasn't there talks were moved up but at the end of the day we still ended over an hour late.

But dinner wasn't the reason yesterday was so good. In the morning Scope Man gave his presentation and he mentioned working with Kiwi and me, doing good stuff and that people would get to hear about it in the afternoon. He is very well known and regarded so it is really nice to have him mention not just my boss, but me! Then Kiwi gave his presentation on my work in the afternoon. He did pretty good. Every now and then he would look at me when he wasn't sure about something and I would nod my head yes. He got a couple things wrong but they were minor and no one knew but me. There were several good questions afterwards and some good suggestions. In the acknowledgements he put my picture up and said that all the work was done by me. And since everyone knew what I looked like I had several people come up to me afterwards during the coffee break, then during the reception to tell me how nice the data looked. Kiwi thanked me several times for putting his presentation together. He made a comment that it was short compared to the others, but I reminded him that we made his presentation to fit the time allotted. He didn't run over like everyone else. The reception was nice, it was on the river with drinks and appetizers.

The only not so great part of the day was that I had to go to work after it was all over. I had to finish processing my PBMCs.

Than the day ended with the bus driver leaving me off in front of my apartment. The stop is actually several blocks away but since I was the last person on the bus he dropped me off on my doorstep!

What a week.

There is a departmental retreat tomorrow in a department that Kiwi holds a secondary appointment. He is giving a 30 minute presentation about all of my dissertation work. As a result I didn't do any bench work this week. What Kiwi doesn't realize is that it takes a considerable amount of time to analyze and prepare my images and movies. I make live-cell microscopy movies. If I do lab experiments everyday there is no time to prepare data. The ideal would be to do an experiment then spend the next day going over the results. But as we all know experiments must be done everyday until I'm done. So I had all of this backed up imaging to prepare and it took me all week. This week was just grueling.

Tuesday night I stayed until 12:30am then had to get up early the next day to proctor an exam for Kiwi since he wasn't going to be around. I'm still not sure how I got talked into that! And that day (yesterday) was just crazy. I was all ready to give the exam, I even got there 1/2 early, then the fire alarm went off. Luckily we were let back in early enough that they were able to finish the exam. Then I went back to my building (the exam was in a different building) to find the power was out. Emergency power was on but there are only so many outlets to plug major equipment into. So everyone was running around looking for power strips and extension cords. The lab next to us, FD and I all had samples that had to be processing so we were hooking up hoods and centrifuges and working in the dark. And for some reason, the other buildings had their power restored about 3 - 4 hours before we did. Another odd thing, the air conditioning was on full blast. We were all freezing. It's like all the power was diverted to the air conditioning. Who cares about freezers warming up, cells dying, virus escaping hoods - as long as the air is on! One good thing about the fact that most of the Center is still empty was there were emergency outlets places where there was no large equipment, so I relocated myself to the aisle, plugged my computer into an emergency outlet, and work on Kiwi's presentation. I made the presentation for him because I have all my data and know what I do. Even though it took me what felt like forever I still could do it faster than him. So the day became very long because of the exam then the power outage. I was running so late that I didn't get my scope time in the evening and had to come in early this morning to do it.

And today the building still wasn't functioning properly. Every now and then we hear a big "whoomp" and that either means the power is going out or something is wrong with the air. And today we had the opposite problem - we had power but no air. Some rooms reached over 85F. This crazy building. It's brand new and full of problems.

Not only did I put Kiwi's presentation together but I had to make him a 'cheat sheet' that told him how I did my experiments and what various reagents were used for. And what cells he was looking at. This tells you how little he is involved in my project. I had to make a cheat sheet.

Because of the retreat tomorrow I'm doing part of the monocyte isolation today and finishing it tomorrow evening after the retreat. I tried to talk FD into processing some of it for me tomorrow but he said he was too busy. Oh, well. I tried.

I'd be home by now but I had to spend time with Kiwi putting the movies into the presentation because he doesn't know how. With all this work I put into it he better do a good job!!!

I can't wait to to just veg tomorrow, listen to presentations, eat a good lunch and not be in lab. I talked Ding into coming with me so I'd know someone there. And the new post doc is coming with us. So I guess we'll get to talk to him a bit.

What benedryl does to me . .

This is how I was last night after benedryl:

• Unwrapped two pipette boxes in the same time it took FD to unwrap about 10.
• Put autoclave tape on three boxes, which FD had to go back over on the second side.
• Weaved while walking

Here's the big one:

• Stood in front of my apartment door, pointed my key chain at it, hit the unlock door key. Didn't realize why the house door wasn't opening until I heard my car beep behind me.

In a haze

I finally have a little bit of time to write. It's been hard to find time lately. However, I'm going to wait until tomorrow. I was forced to take benedryl because I was having a reaction to something. My eyes and lips were itchy and my face was hot and all red. It has taken me way too long to type this much. It took me five tries before I even spelled the title correctly.

Worth reading

For those of you who don't frequent her site, I think Jenny F. Scientist's post "Dear First-Year Grad Students" is worth reading. I want to print it out and give it to all the rotation students visiting our lab. But I will resist the urge, I hate to be the one to dull their enthusiasm.

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!

"America's Most Smartest Model"

This is the title of a new reality series on VH1. This annoys me. It was probably done purposely, to poke fun at people, but come on. This title makes me want to not watch it. And it belittles anyone who is actual on it. The winner should be the first person to correctly explain what is wrong with the title of the show.

Rugby, a new sport for me to watch?

I have been exposed to more comments on rugby in the past week than I have my entire life. I love football. I love the NFL network where there are football things 24/7. I love how physical it is, the pounding of bodies, the athletic feats, the passion of the players and fans. But I've never watched rugby.

StyleyGeek recently had a couple posts talking about rugby. And cluelesslee has talked about rugby and enlightened me to the fact that the US is in the Rugby World Cup. Wow. I had no idea. Rugby is never in the news here. I'd like to watch it on television but I'm not even sure it would be on a channel that I would get. Is it even broadcast in the U.S.? It has to be, I'll have to look because I haven't yet.

I know nothing about the rules of rugby, it looks similar to football, but just without padding, but is it really? For those of you who know rugby you'll have to excuse my ignorance but I'm out to learn more now.

I also find it interesting that sports which are very popular in the rest of the world, like rugby and soccer, get very little notice here in the U.S. - just an observation.

Lab photos

My boss gave a seminar earlier this week which included data from research that FD and Mo were and are working on. Okay, no big deal. And you know how presenters acknowledge people who contributed or helped out or whatever at the end. Also no big deal. One thing I have to give me boss, when he presents data that a student or technician produced he gives them credit and usually mentions that person as the data is being presented, not just at the end of the presentation. (Of course, unless it's LB's work, when he gives him credit for stuff he didn't do. But I digress.)

At the end of the seminar Kiwi popped up a photo of the entire lab. Yes, everyone who works in the lab. Admittedly we are a small lab but still, he went through everyone in the photo and said a sentence about what each of us worked on. But it was an old photo that had our lab manager in it who he let go. He fumbled when he got to her and said something like "This is Lab Manager, she's not with us anymore. Actually, I'm not really sure what she's up to nowadays." He went through us and forgot the acknowledgement page of the people who actually contributed until after the applause when he was flipping to the last slide.

He also redid his faculty web page and put all of our photos on it. Close-up individual photos. I kind of wondered why he insisted on taking a close-up of me by my poster at the departmental retreat, like less than 3 feet away close-up. I couldn't get him to back up even when I tried. He said he wanted me by my poster but the picture was way too close to fit the poster in.

A couple days ago I walk by his office, which I usually avoid, and he had those photos hung up on the outside of his door. He always hangs stuff on his door (the outside of his door) - seminar announcements, meetings, pictures his kids made - but this was a first. It struck me because it comes across as if he's proud of who is in his lab and wants everyone to know. None of the other PIs have pictures on their doors.

It also struck me as a bit odd. Anyone ever notice photos of lab members that their PI takes anywhere? Like on the outside of their doors?

UPDATE:
Fiance said those are mug shots. So if anyone finds us wandering around we are returned to him.

What's up with my boss?

Today I had to process blood in the hood room of another lab in the Center. It was a bit of a hassle because of the crazy lock mechanism on the doors (it takes two hands to open). I had to lug all of the reagents and stuff I needed from one hood room to another all afternoon. And after I started processing I realized that the other lab's centrifuge didn't refrigerate. It only worked at room temperature. I thought I was going to have to spend the afternoon running between three hood rooms because ours was full, I was working in one, and there was a refrigerated one in another hood room. I couldn't use their hoods but I could use their centrifuge. Kiwi was there when I was frantically running from hood room to hood room to hood room. He decided to be useful and told me that the lab used to process blood at room temperature all the time. So, that's what I did. The cells looked okay, just a little haggard at the end of 4 hours but they were alive. I'll see what they look like tomorrow.

While I was starting to process blood, immediately after the centrifuge temperature conversation we had this conversation:

Setting: Me preparing to start blood in the hood, all gowned and gloved and ready to go. Kiwi standing on my side of the BSL2 room door with no protective clothing. Trying to talk to me. I use this as an opportunity to mention the other hood room.

Me: So, did you talk to Incompetent Lab Manager about getting our other hood recertified?

Kiwi: Yeah, last week I sent her an email about it.

(Silence . . . me staring at him saying nothing.)

Kiwi: (hesitantly) Well, I could go ask her again about it. See if we could get things moving along.

Me: (in a tone that says, yeah, you better.) That would be nice.

Kiwi: Okay, I'll get onto that.

Me: Thanks.

(Uncomfortable silence.)

Me: When I have some free time next week . . . well, not really free time, don't have any of that. . . I guess when I have spins and incubations I'll get the other hood room set-up so when the hood is recertified the room is ready to go.

Kiwi: (enthusiastically) Okay, that's sounds good. Let me know when you're going to do it and I'll help you.

Me: (slightly stunned, no words come to mind immediately so . . ) Okay, thanks.

Kiwi: (smile on his face) No problem!

(He leaves the room and I get to work, a bit baffled)

Now he's really starting to scare me. I mean, someone mentioned to me the other day that maybe he's medicated. And in all honesty, I think he might be.

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.

Back to blogging

I've been absent for awhile. Not for lack of having things to talk about. In fact, things happened at our departmental retreat that I feel compelled to talk about because they're just too strange and disturbing, but I'll get to that at another time.

I've been very busy at lab lately. I've been at work early, around my normal time, but I've been working later than usual. I've been getting home after dark and I either eat and go to bed, or just go to bed. Therein lies the problem, even though it really isn't a problem.

I realized that when I look at my favorite blogs or write posts on my blog at work I tend to get sucked in and don't accomplish what I should in lab. I have papers I could be reading, data I could be analyzing, planning experiments, working on my dissertation, looking for jobs - the list goes on and on. And I realize that even if I didn't get lost in the blogs the list would still never really disappear because new things always develop.

Anyway, I decided that I wouldn't do blogging at work. During incubation times I would try to be productive. Yes, productive. But as a result, I didn't blog at all. By the time I got home I was too tired and there was no way I was going to turn on a computer and do anything.

So, to heck with that idea. I enjoy blogging. It's a way for me to release stress and share the craziness that occurs around here. It's a way for my mind to take a break because I can't work all day long without stopping. I have a tendency to go and go and go and I often don't even fit lunch in until after 3pm or 4pm, if at all.

What I decided: I'm going to blog whenever I darn well feel like it. I just need set time limits on my browsing and not get sucked in.

So the mystery picture is. . . .

I'm very impressed with mrswhatsit's guess. The mystery picture is actually a chandelier made of glow sticks in a tent. Since it was pouring when we were camping she decided to break open the glow stick and make a chandelier. And we were in different tents so she sent the picture to my phone. A memento to remember the evening by.

Espresso machine

I recently remembered that I had an espresso machine. It's been sitting on my kitchen counter collecting dust. I haven't used it because I couldn't find the instruction manual and I didn't want to blow it up.

Last week I went online and after some searching found a manual for an espresso machine similar to mine. That's all I've been drinking since then. I haven't made regular coffee in awhile. I'm assuming that the number 4 on the glass container means 4 shots? If so, I've been having 4 shots of espresso every morning. It's been a good jump start since I've been so tired lately. But nooot necessarily good for me. Has anyone ever heard of people developing tolerance to caffeine?

I'm being treated like a human being.

An interesting thing happened today. It's been happening more and more lately. The Kiwi is talking to me like I'm a human being. Not only that, but a human being that knows what it's doing. He actually wants me to do something not related to directly to my project. This is the first time all the years I've been here. Yes, it will benefit him (it's something for a new grant he's writing), but it's still amazing. And he's been talking to people about my work. Good stuff. He's even presenting all of my data at the Departmental Retreat of the other department he's a faculty member in. I just found that out today while he was bragging to the new rotation student about what I do. He told her I'm "dangerously close" to finishing up.

I'm not really sure how to act when he's like this. The best thing to do when he's like this is just to go along with it, because it can change from day to day. It's happened before. It's a calm before the storm. FD seems to think, he says he has a feeling, that Kiwi is trying to be a better boss. I don't know. I think it's a phase. This has happened too many times before. He's only in a good mood because he likes what my project is doing right now. And he's using my cells and my assay to get data for his grant.

But it does make me feel good that he likes what I'm doing right now. That doesn't happen often.

Mystery Picture

My sister took this picture on her cell phone this weekend. Can anyone guess what it is?

I had a dream

So, this first part isn't a dream. I'll get to that.

This weekend Fiance and I went camping with my sister and her boyfriend. I wasn't sure how it would go because we don't always get along. But it was one of the best times I think I've ever had. It was better than I could have hoped. It was a peaceful, private area next to a creek not far from where my parents live. I was so relaxed. Work only popped in my head a few times but didn't stay long. It poured all night, there were two waves of thunderstorms. The storms were so bad that the thunder shook the ground. The water level went up almost a foot. But despite the weather I had a wonderful time. I don't remember the last time I was that relaxed.

And I must say, that with weekends like that I can handle the work week a lot better. I'm not nearly as frazzled as I normally am, even though I have several deadlines that I'm not going to make right on time.

So to the dream. I dreamt that Fiance and I were camping in another country. I don't know which one but it felt like New Zealand because my boss, the Kiwi, eventually was there. Wherever we were, a rhinoceros appeared and it sprayed our tent because it though our tent was another male rhinoceros. It was going to attack so we had to run out the back of the tent. Then all of a sudden we were in another tent in another area. And my boss was having a meeting somewhere near because I saw him, but he didn't see me. While Fiance and I were in the tent Scope Man, the member of my committee who I do microscope with, was trying to look through, not in, but through our tent with a new portable microscope he had. I saw him and yelled at him. I demanded to know what he was doing and he said if I would let him publish his data then he would tell me. Then I woke up.

I know. I'm pretty sure I need some help. I'm not even gonna try and figure out where that one came from. But my dreams about work are getting weirder and weirder. Anyone else have really odd dreams about work?

Another meme to distract me!

I've been tagged by Dr. Brazen Hussy. She tagged me awhile ago but I've finally gotten to it. Sorry it took so long!

The Four Things Meme

Four Jobs I Have Had In My Life:
Roller skating rink guard and concession stand person (that counts as two)
Student office assistant
Wild animal keeper

Four Places I Have Lived:
Really really small town, Central Pennsylvania
Close-to-bad-part-of-city apartment building, Pittsburgh
Closer-to-bad-part-of-city townhouse, Pittsburgh
Relatively peaceful almost suburb apartment, Pittsburgh


Four of my favorite foods:
Pasta and sauce (any pasta with some kind of marinara sauce)
Sushi (eel especially)
Shrimp (cocktail, scampi, steamed, grilled, etc.)
Waffles and gravy (my grandma's version with chicken)

Four Places I’d rather be right now:
Private campsite near my hometown
Bed
Wildlife Refuge in the Ozark Mountains, Arkansas
My parent's house

Four movies I can watch over and over:
The Muppet's Christmas Carol
Speed
The Prince of Egypt
Dante's Peak

Four TV shows I like to watch:
CSI (Las Vegas, the original)
Bones
The Closer
Saving Grace

Four websites I visit daily:
Weight Watchers
Bloglines
Twitter
iGoogle

Four early musical influences:
Bon Jovi
Madonna
Def Leppard
Chicago

Four Computers I’ve Owned:
Compaq laptop (can't remember what kind)
Dell Desktop (ditto)
Dell Inspiron 600m



I tag:

Geeka507
Kelly
Jenny F. Scientist
mrswhatsit

(See Geeka, I resisted the urge to tag you. So you don't feel obligated to do it!)

To Steamed Puddings

Kelly,

I think I'm doing something wrong on your site. When I leave comments I do it under the anonymous setting since I don't have an account with any of the ones where you can log in. I don't think you've been getting them. Let me know what to do to leave comments. I replied to that wonderful post that mentions all the different websites you found.

Thanks!

Perverse satisfaction

After yesterday's episode, FD informed me that LB's flow staining that he did yesterday didn't work. He used several antibodies and all he got was autoflorescent background tails.

So sad. I can be spiteful, I know it's wrong, but sometimes he gets me to the point where I just don't care.

I'm going to bop him in the head

He pushes my buttons, I admit it. He just plain pisses me off when he speaks. So it is just compounded when he's speaking to me and arguing about my methods.

Our poor lab tech Ding is trying to learn too many things at one time. She getting the procedures confused and mixed up and she knows it. But the Kiwi wants her to learn everything as fast as possible all at the same time. LB is teaching her a couple things, one of which FD and I are going to have to reteach her, namely flow. But she's trying to do flow and elispots at the same time. She offered to work late tonight to get it all done so the work she's doing fits into his schedule. He's not the one with a 2 month old baby to take care. He's not the one with 10,000 things to learn in two weeks. So I told her if she wants to do the flow staining tomorrow she can. Just fix the cells, put them in the fridge, and stain them tomorrow. It will be fine, she's only doing CD14 staining.

LB stood there and told her it was a bad idea. I told him it's perfectly fine. He told her it isn't. Notice he's talking about what I said, disagreeing, to her not me. Which pissed me off even more. Then he said it's "bad practice." I almost decked him and called him a f#$% a$%^hole right there. I said I do it all the time, my stuff is fine. He told her it's bad practice. I finally looked at Ding and said both procedures are perfectly fine, it just depends on what you're doing and what antigen you are looking at. Neither way is wrong and neither way is bad practice. Then I walked out.

I am soooo pissed I'm shaking. I know he shouldn't upset me so much but he's an arrogant bastard. Someone who leaves antibodies on for 2 hours to overnight so his staining looks better, who counts background as events, who doesn't even have the appropriate controls needs to keep his mouth shut and learn from someone who does it all the time. FD called me a flow guru, it's true, I need to calm down.

But I really want to hit him. He better not speak to me again. That's the first time since the email episode awhile back (part 1, part 2)and all he does is act like an a#$%hole.

I tried to stay dry

Yesterday I had a lot of computer work to do. The weather was crappy so I figured I'd stay home and work until the rain let up a bit. I didn't want to stand at the bus stop in the pouring rain. So I waited, it slowed, I went to work fairly dry.

Then I went home. FD and I caught the bus at the same time (we live near each other) and the sky was clear - it was cloudy but wasn't raining.

The closer we got to home the weather rapidly changed. The sky got darker, the wind picked up, rain started to fall, then rain started to POUR. I mean a torrential downpour. It wasn't lightening though, until I commented that at least it wasn't lightening. FD wanted to hit me, he's a strong believer in Murphy's Laws.

I had an umbrella but it didn't do much good, it kept my bag fairly dry. And thank goodness Fiance bought me a raincoat a couple weeks ago. There really was no point in running because I had to go several blocks, so I would be just as wet walking. Running actually splashed the water more. I had three streets to cross and the water was just rushing down them. On both sides of the road water ran about 3 feet wide and several inches deep, more than enough to go over my shoes. There was no avoiding it. And water pooled on the sidewalks.

So, point of the post - even though I stayed dry in the morning I made up for it in the afternoon. My only pair of sneakers are still wet today, it was like I dunked them in a bucket of water. Here is what I looked like when I got home. The water soaked the whole way up to my shirt. Admittedly, I'm only 5'3'', but that's still pretty high.

A computer-free weekend

I actually managed to relax and stay away from the computer all weekend. I thought about email, thought about blogging, but managed to not log on, not even turn on a computer until right before I'm climbing in bed tonight.

Fiance and I decided to take on a new hobby/recreation. As you can see from my profile and am a country girl temporarily stuck in the city, but slowly getting out. I moved out of the near center of the city to a more suburban area. I just need to keep moving out, even though I do like the neighborhood I'm in now it's not the same as having no neighbors and no traffic sounds.

Back to the hobby - it's camping. Camping is something I've never done but always wanted to. Don't ask me how a country girl made it all her life without camping but I did. I just wasn't something my family did on the farm. We lived in the countryside, so why did we need to go out into it at night. Also a major contributing factor - my mom hates it. Fiance and I thought that it would be a relatively cheap thing to do on the weekends to get away from things. Yes, there are initial expenses that we splurged on for the gear, but at this time of year we got almost everything for over 50% off. And after the initial purchases the most common expense is food and batteries.

In a week I'm going camping with my sister and her boyfriend. In the past year it is something my sister has taken up and she is actually excited about camping with us. This means a lot to me because her and I are two years apart, she's younger. And we never really got along. However, we get a long much better that we live in separate houses. We may have actually found something that we both can do together.

This weekend Fiance and I decided to test the new equipment in a more 'controlled' environment. I wanted to make sure everything worked before we went out into the middle of the woods. I envisioned the tent collapsing or something. So we camped in my parents' backyard, that way if anything happened we could go inside. I know what you're thinking, it's not really camping if you're in a backyard, but you don't know our backyard. There is a small woods lining the property and acres of farmland, and no other houses are visible from the back. And all sorts of wildlife traverses through it. We commonly have raccoons, deer, skunks, sometimes opossums. Bears have even been spotted, not just wandering through the yard but tearing down bird feeders attached to the house. So even though it's in a yard it's still an experience. And what's nice about it is that we get to visit with my parents and still have time to ourselves outside.

So after watching the first half of the Steeler game we went outside. Fiance had a nice fire going with wood that my dad got from my uncle. We must have spent an hour just sitting there, listening to the sounds of the forest, staring at the fire. Looking into the flames is very hypnotizing. Though I must admit that a few times lab popped into my head and I had to force it out. It was so nice in the tent all night because it was like being in the center of a sound spa. I have a sound spa by my bed that I listen to every night to try and distract me from the noises outside, and also to drown out ringing in my ears. It was just fabulous to have it in stereo!

Everything worked fairly well except it rained pretty good during the night. Which actually turned out to be a good thing. The tent leaked. It wasn't a bad leak, it's not like we were swimming in water when we woke up, but there were small drops of water that had collected in tiny puddles. Water had leaked in through the seam above the leak guard on the door. I'm going to see if I can take it back since it's the first time we've used it, if not, it's a Coleman guaranteed for 5 years and they will either fix it or replace it.

What matters is we had a good time, relaxed, and got out of the city. I can't wait to go again!