Today was a bad day at the lab for me ('typical', according to Danielle). Experiments didn't work, I freaked out (again) over my results, got depressed about my chances of publishing a paper and I didn't see my PhD ending this year.
My experiments stopped working about a month ago. I've had it before; it seems to be an up and down motion of productive weeks and unproductive weeks. I think in-vivo patching is a precarious interplay between lots of factors. Get one wrong and your experiment will fail. Get them all right and you have a chance at results, provided you work hard.
I aim at patching in the cerebellar nuclei, but recently I often overshoot them and get vestibular nuclei instead (which seem to be very easy to patch for some reason). Also, the patches I get in the cerebellar nuclei are of bad quality. I get to 100-200 MOhms of seal and then they drop off. Or they just don't open nicely and I have to dump the recording because nothing can be learned from it. It's probably something to do with slight differences between mice and a slow drift of the stereotaxic location of the nuclei between generations of mice. Why the patching is so hard, I don't know. Tomorrow I will have freshly polished and flamed electrodes. I will throw out my internal solution and make some new. Hopefully this will solve some of the problems.
I freaked out over my results because I feared I might have patched a lot of vestibular nuclei cells. By inspecting my data closely, this turned out not to be the case. Thank god, I might have been forced to throw out months of work... This of course caused my slight panic attack over my chances of publishing a paper and finishing my PhD. When the attack was gone I decided to take matters into my own hands and have another try at an experiment (for the result, see above).
Fortunately I have very considerate and wise colleagues. One particular in-vitro patch clamp colleague (who always plays creationism vs evolution or religion vs atheism debates for the lab, which we all thank him for ;-)) had very helpfull insights: A PhD defence shouldn't be too much a landmark event. It doesn't define you as a scientist, it's merely a thing that you need to do to make life easier. It's something you need to pass to get to the next level, but it doesn't define your expertise or you being a scientist. It's just 'a thing'. The most important issue here is to publish thorough papers that you can always defend. So, my PhD book will be more of a 'booklet' with one or two papers and unpublished chapters. Which might end up as published work someday. Don't get me wrong, I would be very happy to get the definite proof on some of the things I've been working on, but it's just not going to stand in the way of me moving on.
So, tomorrow I'm going to talk to 'my boss' to talk about a PhD defence date for this year (2012, remind me if I missed the mark ;-)). I will continue work on my cerebellar nuclei and cerebellar cortex stuff. If it all works out, it will be submitted or published when I'm finished. If not, so be it.