Sunday, February 12, 2006

Laboratory Blunders

It’s refreshing to know that, regardless of how much experience or training we undergo, we remain human and still make stupid mistakes. I’m not just talking about fighting with laboratory equipment which hinders experiments or doing too many things at once to the point where you forget what is going on, but the type of mistake that is so rudimentary that you can’t help but slap your forehead and roll your eyes. At points in my scientific training career, I will admit to being the poster-girl for stupid laboratory mistakes…you know, things like forgetting to, err, add DNA to your transformation, and then standing around confused as to why the agarose gel does not look as expected (me? do something like that? never!). This time, however, the error fell in the hands of my PI and Assistant PI and, like I said, it’s nice to know that even my superiors are still error-prone at times (particularly since I am sure that in my next way-too-many years, I will make similar easily-avoidable mistakes, and I know that my PI can just sit back and smirk but know that he has been in similar situations).

To make a long story short, the assay detects hydrolysis by looking at the amount of free phosphate. My PI had the idea that the protein may be more reactive if dialyzed into a different buffer solution, and discussed this with the Assistant PI, who is apparently the protein-master of the lab. Ultimately, they decided to do the overnight dialysis into PBS… otherwise known as phosphate buffered saline. That’s right, phosphate. This didn’t dawn on my PI until the colorimetric detection agent was added, and instead of a barely-detectible-by-human-eye color change, the solution in the well-plate turned instantly dark green. After a few moments of looking confused, suddenly it hit – in his case, with a response of “Oh, DUH! What were we thinking?!” Yes, the laboratory lesson of the week was, in the event of attempting to detect hydrolysis of ATP by measuring the release of phosphate, it just might not be a good idea to use a phosphate buffer! Luckily, this problem was easy to fix within 24 hours by dialyzing other fractions in a non-phosphate containing buffer, but it still provided a bit of humor throughout the lab for the rest of the week.

In other news, it has been a weekend of studying, as the biomedical graduate students are facing our first exam of the semester tomorrow. I’ve been remaining so hopeful for a snow day tomorrow which would push the exam back to Wednesday, giving me two whole extra days to study and therefore somewhat of a chance to pass. We did get about 8” of snow and knowing how inept D.C. is at dealing with snow, I thought for sure we would at least get one day off, although things seem to be clearing up well, particularly for a city which goes into panic-mode with just a few flurries. The majority of school districts in the area have already closed for tomorrow, or at least declared a two hour delay, so I’m still keeping my fingers crossed for two hours… that’s all I need! Just a two hour delay – I’m willing to go into lab and class for the rest of the day… is it really too much to ask for just two hours off? Or even a declaration of “any exams cruelly scheduled for 7:30am despite the fact that the course meets at 9, in order to allow enough time for completion of the exam, are hereby postponed until Wednesday.” In the meantime, it’s back to studying the nine bazillion different types of collagen… and checking the snow cancellations every fifteen minutes.

Monday, February 06, 2006

SUPERBOWL XL CHAMPIONS!

Pittsburgh Steelers -- Super Bowl XL Champions! And my beloved Hines Ward as MVP! Seriously, I don't think I've stopped smiling once today. What an amazing post-season journey it has been, considering that after the Week 13 loss at Cincinnati, I didn't even think they would make the playoffs. And then to beat Cincinnati in the wild-card game, Indianapolis in the divisional round, Denver to actually win the AFC Championship game, and then despite an absolutely horribly played first half... to win the Super Bowl. HERE WE GO, STEELERS, HERE WE GO!


Saturday, February 04, 2006

The Value of Time

When it comes to my first laboratory experiences, I'll admit that I was pretty spoiled. My first research experience outside of undergraduate course-related bench work was at The Institute for Genomic Research, one of J. Craig Ventner's well-funded research institutions. During my ten months as a research intern, I was taught that time was considered to be more valuable than cost, in the sense that if one method was pricier but took less time, it was a worthwhile investment. While there was a limit to the amount of funding available, when it came to everyday things in the lab, it was no problem. Instead of running agarose gels which can take some time depending on the size of fragments, we used a BioAnalyzer chip. When preparing samples, I was taught to always prepare more than actually necessary -- after all, if one of your Eppendorf tubes somehow is knocked over, you'll have a spare, instead of having to go back and repeat everything the next day. In addition to funding, we were spoiled in terms of general lab maintenance. If glassware was used, it was just placed in a plastic bin, and the next day it was back on the shelf, sparking clean and autoclaved. In fact, it wasn't until I started my current laboratory rotation that I even actually saw an autoclave with my own eyes (don't worry, I am completely despoiled now, and am a pro at autoclaving my own glassware and all the trash from the lab). My time at TIGR was, to date, the best laboratory experience I have ever had, not just because of the funding, but also because I truly enjoyed what I was doing, and the Howard Hughes fellow under whom I was working took the time to actually explain things to me and let me do some thinking on my own. My senior year as an undergrad, I did research at the National Cancer Institute. While not funded to the extent of TIGR, the lab still did not find itself pinching pennies at every corner. Once again, the general attitude seemed to be that time was considered to be more costly than supplies -- after all, if you can get more experiments done and generate more data, then there is a better chance of securing a grant.
Then I entered graduate school. My first rotation was in the best-funded research laboratory of the entire university, so the experiences were similar to my previous ones. The expensive equipment was available for my use, even though it would be much cheaper to do the same experiments by hand. My second rotation (the lab in which I will be staying for my thesis), however, is slightly different. It's not that the lab has no money, but just that it is much more cost-conscious than any other in which I have worked. Over the past two weeks, I have been busy attempting to make sense of a GST-CBP plasmid sent from another lab, which is the positive-control in an RHA pulldown experiment that I am doing. However, the lab did not send a map (and actually apparently couldn't 'find' the plasmid either, so they just sent crude cell lysate) and so, after a few literature searches, was able to identify the vector used for the plasmid and the amino acid sequence of the fragment of CBP. After generating what I thought was a correct plasmid map, I went to work doing a few restriction enzyme digests. Using my training from previous labs, I made a larger reaction mixture than necessary. Although a mini-gel will only hold 10ul, I used 40ul for my total volume reaction, just out of habit since this was what I was originally taught. The fourth-year student in the lab, who has truly been a savior in terms of helping me with questions and not laughing or calling me stupid when I do things incorrectly, pointed out that if I was only going to run 10ul on a gel, and the only purpose was to check the size of the fragments and I wouldn't need the digested pieces for anything else, it was wasting both the DNA and the expensive restriction enzymes to make more than necessary. Following his advice, I started using a 10ul total reaction volume. Naturally, I pour my a gel for the thousandth time, let it solidify, load it up, turn on the power source, set it to 90V, watch for the bubbles, and walk away. Twenty minutes later I return, and the gel is floating up off the bottom of the gel box, and all my DNA has somehow spilled off. Since, for the first time in my life, I only made the exact amount necessary, I was stuck repeating the digestion. Thankfully the whole process only took about another three hours, but somewhere, the biomedical research gods are looking down on me and laughing.

Also, for those of you who need a little help getting energized before the big game, might I suggest (complete with sound, so don't forget to turn on the speakers!):
The Original Version
The Playoff Version

Wednesday, February 01, 2006

Student Health Stereotypes

This may be sharing a bit too much personal information, but honestly, today was just too ridiculous to not discuss. In a bit of background leading up to the story (and the part where I may share more personal information than you wish to know), I no longer produce enough progesterone, so without taking progesterone, I don't menstruate. Frankly, as long as it's not causing any other problems, I can't say that I mind at all. However, in order to ensure that there are no other problems as a result, at the end of December I decided to put my university-provided health insurance to use and head down to Student Health. After drawing about a liter of blood for testing (not kidding) and running a bazillion tests, I was referred to radiology for two types of ultrasound to rule out adrenal or ovarian tumors.

This afternoon, I went back to Student Health for a follow-up appointment and to discuss the results of my ultrasound. This is where the story begins.

My Afternoon at Student Health, or Why Every Stereotype You Hear About Student Health is True. A real-life experience in Two Acts.
Location: Examining Room.
Enter: Nurse.
  • Nurse: "What brings you here today?"
  • Julie: "I was told to make a follow-up appointment to discuss the results of an ultrasound."
  • Nurse: "And what were the results?"
  • Julie: "I don't know, that's why I'm here."
  • Nurse: "But what did the ultrasound say?"
  • Julie: "I don't know. The radiologists didn't tell me anything at the time of the testing."
  • Nurse: "So then why are you here?"
  • Julie: "To find out what the results said."
  • Nurse: "So are you in any pain?"
  • Julie: "No."
  • Nurse: "Then what is the nature of your visit?"
  • Julie: "A follow-up appointment. I was told to schedule an appointment."
  • Nurse: "For what purpose?"
  • Julie: "To go over the results of my ultrasound."
  • Nurse: "And what did the ultrasound indicate?"
  • Julie contemplates banging head against wall until she has a severe headache, thereby allowing her to state that she is, indeed, in pain, which will appease the nurse.
Exit: Nurse
Enter: Doctor (but not the one with whom the appointment was made)
  • Dr.: after flipping through chart briefly, "So, the problem appears to be that you do not ovulate, is this correct?"
  • Julie: "Yes."
  • Dr.: "Are you pregnant?"
  • Julie: "Is it possible for me to even get pregnant considering I don't ovulate?"
  • Dr.: "I understand that you are a biomedical PhD student. Therefore, you clearly are not in comprehension of basic human physiology. When you are pregnant, you do not ovulate."
  • Julie: thinks to self, 'no really? When did 'PhD' become synonymous to 'moron'?'
  • Dr.: "Are you pregnant?"
  • Julie: "No, I am not."
  • Dr.: "Are you sure?"
  • Julie: "Yes, I am positive. Last month when I was here, I had a pregnancy test and it came back negative."
  • Dr.: "Could you have become pregnant in the past month?"
  • Julie: "No. Especially since I don't ovulate, which I do believe is a requirement for pregnancy."
  • Dr.: "Why are you here again?"
  • Julie: "To find out the results of my ultrasound. And considering it was a renal, ovarian, and uterine ultrasound, I feel like the radiologist may have noticed if I was pregnant."
  • Dr.: ponders this for a moment, then replies with "Before we discuss this any further, you need to take a pregnancy test. You will find urine sample containers in the restroom."

Like I said, today just proved that every stereotype about every university student health center is, indeed, true. Oh, and in case you were wondering - the pregnancy test came back negative. Quite the relief - clearly, I was reeeeeeally worried about that.