I look at my schedule and groan. First thing - 6 am. End of the last appointment - 11 pm. Every day this week, every week this month, every month this year, and last year, and the year before that, it looks like that, row after row after row of experiments and meetings and conferences and more experiments. Hybe, it says. Incubation. Fix. Refix. Wash. Prehybe. Make electrodes. Wash. Make new probe. Alcohol dehydration. Wash. Make buffer solution. Find a better buffer. Wash. Wash. Wash.
I sigh, close my laptop, pull on gloves. New page in the lab notebook, just like the last hundred-and-seven pages and the last three notebooks, flipping past the taped-in microarray results and quantification data, the photos of electrophoresis gels blank and full, the sequence strings, the charts of restriction sites, the scrawled notations of failure and bewilderment and (all too rare) triumph. 2011.10.28 - Whole mount ISH, H. vulgaris, 24-hr budding, PaxB.
One more time, I think.
My phone buzzes, the text message flashing “Drinks tonight?”
“Not tonight,” I reply. “Late night in the lab.”
I pick up the pipetter and my wrist cracks. I roll it slowly, feeling the tendons twitch and pull, tendonitis I can't let rest. The sun sets outside the windows. The computer in the corner reads off the time as hours flicker by.
One more time, I think. One last run. One more batch. And so it goes - fix, refix, wash, dehydrate, rehydrate, make more electrodes, make more probe, wash yet again, over and over and over.
One more time.
Three little words for three big letters : P. H. D.
One more time.
Author's Note: Some explanation of what I'm doing here. Feel free to read this or not - it is an attempt to explain mostly what is going on in the entry - the science-speak, but also why I wrote it the way I did.
My PhD project involves the evolutionary roots of vision in a so-called 'primitive' system, the hydrozoan Hydra vulgaris. (That link is to a picture I took of one of mine.) If you think of a sea anemone, then scale it down much smaller and a little bit simpler inside, that's a hydra. They have no eyes or eyespots, but they can not only sense light and darkness, but differentiate between colours. My project is to figure out the how and why.
It's a combination project, part recording neural responses - sticking very tiny electrodes into nerve cells - and part determining where and when certain genes thought to be involved in building visual structures are turned on and in use. That latter part is what I describe above. It's a technique called in-situ hybridisation (ISH) that uses a dye-tagged probe that binds to the mRNA, the intermediate stage between a gene (the instruction for building a thing) and the protein (the thing that gets built.) This lets us see blue stain when and where the gene is on. It's a laborious and many-stepped procedure; normally, I stay away from all the scientific jargon, but I was hoping it would be helpful here to see the complexity.
Let me know what you think!