A few weeks ago our latest paper was rejected from the Journal of Biological Chemistry (‘JBC’). It had been sent to two reviewers who each felt that the paper was unsuitable for publication in its current form without further experiments. This is quite normal and is part of the peer review process. You are then given about 3 months to revise the manuscript with the new data and respond (argue politely) to the reviewers comments. If your revised manuscript is still not deemed to be acceptable by the reviewers then it is rejected from the journal and you are then able to submit it to another journal, usually lower down the pecking order.
Yesterday afternoon I made a throw-away comment: “So just two more experiments – one more week – and we’ll be all done.” I shouldn’t have said it – immediately afterwards I tried to take it back, to apologise, but it was clearly too late. Never say “Just one more experiment” or “Just one more week.”
In my current experiments I use semi-automated microscopy to analyse the dynamics of signalling pathways downstream of the Gonadotropin releasing hormone (GnRH) receptor. I use assays that have been used in the lab my me for more than a year, and by my predecessors for many years before me. These assays work.
Today’s experiment was set to be beautiful. I used all new stocks of reagents wherever possible. My mind was clear when I carried out the complicated stimulation time course. Carrying out experiments for me is a bit like drawing: if I’m tense, stressed or pre-occupied all I achieve is a little knot of pencil lines, some grainy, dead cells.
As the InCell Analyzer chugged through the imaging, one field at a time, I hovered around waiting to see the results as soon as they were ready, filling tip boxes ready to autoclave (the downside of having an entire lab to oneself is having to do all the lab chores). I couldn’t believe my eyes when the images were completely blank – well not completely blank – there was ‘background’ staining (non-specific, low-level staining) but the cells had clearly not been stimulated by GnRH.
How could this be? This has never happened before! In those last-ditch experiments one goes all out to get the perfect results, by scaling up to a larger experiment than before, by changing all the reagents in favour of nice new ones? Those fateful words stating the end is in sight.
Sadly, as I have experienced all too many times, the end is rarely as close as you think it is. My suspicion is that the new batch of hormone I made up from powder today was to blame. Perhaps it was a really old vial (and thus degraded), or there wasn’t the right amount of powder in the vial. Hopefully I will be able to make a guess at the answer next week when I run some experiments to decide which reagents I trust and which I should bin, so pushing the completion date for the revised manuscript back even further.
These experiences make us superstitious – the old stock of hormone might have lost some of its activity (it might have been thawed one too many times by the Summer lab students) but at least it works! From here it’s only a short step to using a favourite pipette for every experiment, or indeed, wearing lucky socks.