If running a scientific project is like cooking, then my usual modus operandi in the lab is to prepare a lavish, many-course meal. The meal, in this analogy, is the overarching goal of what I want to understand, and each course is one independent angle on that question. Such a multitasking approach is useful: you get to try out different competing theories, which can answer your question faster, and if something crashes – say you burn the casserole – you can still keep the project afloat with more promising routes and extra helpings of dessert.
My multitasking tendencies got more extreme during my experience as a team leader in a Dutch biotech company – when you have a group of people working for you, it’s so easy to expand your menu. My challenge in returning to science as a re-entry fellow was to adapt to being a lone-wolf chef once again. Although it’s been hard at times, I’ve managed to run my project in the manner to which I’ve grown accustomed, while at the same time keeping it manageable for my one pair of hands.
In the past month, however, I’ve had to change my approach completely. My big screen paper came back from an excellent cell biology journal with three positive reviews, but after a bit of back-and-forthing with the editor, I was charged with performing just one final experiment to flesh out the mechanism a bit more. And I was given three months in which to do it.
Never has my career depended so much on the outcome of one single experiment. Applications for independent fellowships are coming up this autumn, and though I have a good shot at them, I have a feeling that this paper will be what makes or breaks the acceptability of my track record, which will be an important component. So a successful experiment will land the paper into a safe and well-regarded home in time for the deadlines, which will facilitate my applications; in turn, success at winning a fellowship will allow me to stay in science as an independent researcher. Failure, though: I almost don’t want to think about the reverse. No timely paper, no fellowship, no position. In that event, it’s time to do yet another post-doc to try to consolidate my chances – or to finally surrender gracefully to the inevitable and leave research altogether.
So my laboratory life has narrowed down to an incredibly focused point, and I have to say I’m enjoying myself immensely. One key experiment: what could bring more lucidity to my life? It’s like trying to boil the perfect egg. Everything else is on hold as I grapple with the best way to deliver. The editor referred to the experiment as “straightforward”; well, yes and no. There are two ways to do it, neither of which are up and running in our lab: a classic biochemical approach, and a new-fangled strategy involving Förster Resonance Energy Transfer microscopy (FRET). Naturally I’ve called in all my connections and received advice – the old-fashioned biochemists, horrified, tell me to avoid FRET like the plague, while my imaging-savvy colleagues would rather put out their own eyes with a Gilson P1000 than spend all day in a cold room pulling down elusive proteins so fragile, urban myth goes, that if you so much as hold a tube at its base instead of by its lid for a few seconds, the game is up.
Typical me, I’ve decided to pursue both in parallel. Each technique is challenging and interesting, and equally useful to have under my belt for the future. And clearly, doing both will maximize my chances of success.
One month into the three-month period, and I’ve had some modest success with the FRET, though I’m not sure it’s sensitive enough to reveal my phenomenon clearly enough. And it’s gruelling work: preparing the cells and capturing the images is easy, but analyzing the data is painstaking and slow, one cell at a time with dozens of analysis steps in the pipeline. For example, I managed to look at four today, and I’ll require many dozens; rainbow-hot images of phtotobleached cells are seared into my retina and haunt my sleep. Meanwhile the reagents for the biochemical assay have arrived from America, and I’m going to set up a few pilots next week. It’s more sensitive, and I’ve always had biochemical green thumbs, but I’m worried that my process will be too localized in the cell to show up in a mass population of lysed cellular proteins.
So the game is still all to play for, and the stakes are incredibly high. I should be worried and stressed, but instead, I’m upbeat, ruthlessly efficient and buzzing on adrenalin.
It’s going to be the best damned egg you’ve ever eaten.
Good luck with that egg, Jenny. I’d be blasting it with that FRET laser right about now 😉
How long should I photobleach an egg to get a nice runny yellow but firm white?
That would be the (in)famous 65C egg then I guess 😉
Good luck with the biochem assays as well the FRET!
So, after all the to-ing and fro-ing around the base camp and occasional sashays up the South Col, this is the final assault on the summit.
By God, Squiffy, I Wish I Was Going With You.
Since it’s often considered bad luck to wish someone good luck, I shall instead say, in theatrical style:
“Break a leg”…
…or should that be “egg”?
i think that joke has gone oeuf.
Not sure if I poached it from someone else, but perhaps it did get scrambled in the telling.
That’s a yolk to stick in the albumen.
Jenny – what will you do if both experiments work? Write them both up?
Or worse, they both work but give conflicting results…
If they both work I’ll use them both. If they give conflicting results, we’ll have to sit down and decide What Is Truth. I think it very likely that the biochemistry could be honestly negative because the effects I’m looking at are very local. So if I get no differences in the lysed cells but local differences in the FRET, it’s not actually inconsistent or unsurprising. The other way around would be weird, but I really doubt that will happen.
Since I wrote this blog I’ve come up with yet two more ways to go after the problem, so we’ll add that to the mix and see what happens!