alterations and another mystery

For Labor Day weekend I set out to finish revisions for a paper I submitted months ago, cut Joelle's hair all off, do all of the dishes, and unpack boxes that have been sitting in my bedroom since I got here in April. I also intended to get a fairly good chunk of work done in the lab.

At the end of the weekend, what I had in fact done was...play the Sims 2 for a good 12 hours, chill with friends on Saturday and Sunday nights and all afternoon Monday, have ice cream and coffee with coworkers, wash half of the dishes, cut about a quarter of Joelle's hair so that she looks rather insane, write about one sentence for the paper and take out about ten sentences, and rearrange all of the furniture in my apartment.

Hm.

The good news is that somewhere in the midst of the half-hearted haircut and the major change in layout, Joelle seems to be making it into her litter box much better. It is also possible that my presence in the apartment (as opposed to the lab) for much of the weekend contributed to her success. I have also been slipping in some raisins and outstanding grapes when she gets treats for good behavior. In any case, something seems to be working.

Back at work, another mystery exists. In the history of my lab's working with my bacterium, there has only been one or two instances where there has been successful growing of the bacteria on what we call agar plates. They're sort of like Jell-o in a Petri dish. My bacteria only grow when there is no oxygen, and they like very specific growth conditions (a.k.a. what's in the Jell-o). Apparently I am not providing them with what they want, because they are not growing. And that is not good.

The question is WHY. There are several possibilities. One is that there is oxygen in the place where I'm trying to grow the bacteria, and that would make the bacteria die. Another is that the agar plates have not been prepared correctly, which could mean that the ingredients are slightly wrong, they got burned in the process of being made, THEY have oxygen in them, or something else. And it is also possible that there is both a problem with oxygen in the growth chamber and issues with the agar plates.

Slowly but surely clues are coming in. First, the chamber that I thought had no oxygen in it seems to in fact have oxygen. It depends on whether I look at the electronic oxygen sensor (which says there isn't any) or at the low-tech oxygen sensing paper strip (which says there IS). I tend to believe the paper strip since when I put excess non-oxygen gas into the growth chamber and overpressure it so that the rubber gloves that are part of the contraption blow up like balloons and stick out, the gloves deflate back to normal pressure within a few hours. This indicates a leak. Not Joe, being a mechanical engineer and rather handy, stopped by the lab and helped me in this investigation and agrees with me that there's a leak. He's funny: he wanted to figure out in which part of the multi-part chamber the leak existed, and he asked if I had a stethoscope so he could listen for leaks. Why in the world would I just randomly have a stethoscope!? Apparently a screwdriver works just as well, because he used that instead and achieved his diagnosis.

Another clue is that agar plates stored in a separate non-oxygen chamber have a high pH. This is bad since the bacteria don't like high pH and will die if exposed to it. What is kind of annoying is that the chamber in which the pH is wrong actually has no oxygen, which is what I want. But since it has nitrogen gas and not the carbon dioxide that would make the agar plates be at a normal pH, it's useless.

I also realized that the previous work done with the bacteria in our lab was performed in a chamber that detected oxygen using the same oxygen detector that seems to be giving erroneous readings in the leaky chamber. That old chamber also had some odd glove pressurization issues, so it's quite possible that it, too, was leaking and had oxygen inside and was killing the bacteria.

So, at least things are starting to make some sense. The long and short of it all is that there are probably multiple issues going on. Now all I need is one simple way to fix all of those issues. ...Getting that simple way is probably not going to be very easy, though!

Thank goodness I took a class in forensic science in college, eh? I'll get to the bottom of this, and hopefully within a couple weeks I'll have bacteria growing AND all of Joelle's bathroom habits contained in one location. Hopefully I will not need to rearrange all of the furniture in the lab in order to do so...!