It started out simply enough. I wanted to know if the treatment I’m applying to cells was increasing cell death. No problem. That’s easy enough. I’ll just use a live/dead stain.
Disaster thought #1: “Ah yes, this stain of unknown age/origin that I found encrusted in ice in the back of the freezer on lab cleanup day should do the trick!”
Yup, I’ve got this totally under control. Now, it says here that I need to quantify using FACS.
Disaster thought #2: “I did some rudimentary FACS analyses a few years ago during my PhD using a template that someone else made and that I barely grasped, I’m sure I can just hop down to the FACS facility here that I’m completely unfamiliar with and, relying on my superior memory of boring experiments from 36 months ago, will be able to figure it out.”
Luckily it did occur to me that I will probably need to sign up to use the machine, so I check online and find…oh dear. What’s this? A registration form that needs to be signed by my PI who is currently away for 3 weeks? And even after getting the form signed I then need to attend first an orientation session followed by a training session?
Disaster thought #3: “Pffft.”
So next I do what any reasonable postdoc rushing to do things before her PI gets back and asks what the hell she’s been doing for the past 3 weeks would do. I ask around the lab to find out if anyone has been trained to use the FACS facility that can sign up for me.
Disaster thought #4: “HA! Fuck you system! I am so smart, S-M-R-T!”
To my delight I discover that, Antoine*, a senior grad student in the lab, received FACS training about 2 years ago and may have even used the FACS once or twice around then with the help of someone who knew what they were doing.
Disaster thought #5: “I’m sure it will come back to us when we see it.”
Like a good little scientist I then realized I’d better get planning so as to not waste this opportunity. After all, wouldn’t it be better to accomplish everything in one giant experiment so that I don’t further inconvenience Antoine*? I mean, then when my PI comes back I could show her this fantastic complete data set.
Disaster thought #6: “Fuck yeah! I’m an experiment planning machine! I’m gonna do this shit in triplicate and everything! Now where did I put that really expensive reagent??”
Well. Awesomeness of this magnitude takes planning. So I thought of every possible thing that I could ever want to know surrounding this experiment and incorporated it into my protocol (except, of course, the one control I really needed) and I planned it all out, but left myself some calculations to do in the morning since I didn’t have the numbers I needed while I was planning.
Disaster thought #7: “I can do math first thing in the morning, unsupervised.”
I also found that my cells were not quite at the confluency that I really needed them to be, which brings a disaster thought I’m sure we all share.
Disaster thought #8: “It’s still good, it’s still good!”
As the experiment got underway I was pretty sure it was going to be the greatest first attempt ever. I was also pretty sure that though I had never done this before, it wouldn’t take that long.
Famous last words: “I can totally get this done in an hour.”
Accompanying disaster thought #9: “I’m sure the times given in the protocol provided by the company are just suggestions.”
When the big day came to actually do the FACS analysis, Antoine* and I could barely contain our excitement! We ran eagerly to the facility and encountered a bit of a surprise when we looked at the time sheet. Unbeknownst to me, there was a disaster thought #5.5: “We are totally capable of figuring out sign up sheets/times”. It was also right around this time, as I looked around the facility full of people bustling operating machines that I could not identify, that I had my first inkling that any of my thoughts had been disasters.
Disaster thought #10: “I’m sure it’s fine.”
Not wanting to admit that we were imposters who had bucked the system and had no idea what we were doing, we instead just tried to figure it out.
Disaster thought #11: “If you pretend you know what you’re doing, you’ll figure it out.”
Luckily, the woman who runs the FACS facility is insanely competent, patient and forgiving and ending up squeezing us onto another machine (since the one we *thought* we had booked was being used in 10 minutes) and running my experiment for me.
Lesson learned #1: If you’re incompetent, competent people will do your work for you. This is not a good thing.
Well, long story short – for this experiment I now have written in my lab book under the heading “Conclusions”:
Hind sight is 20/20.
*Names have been changed to protect the innocent/equally guilty.