After checking whether those cells I’d plated approximately four hours previous were landing on their intended target area, I left work in the gloaming, but after a few hundred yards, I stopped. And thought. Not about a mosquito-like irritant (because one mosquito is not important), but about the small volume of medium nourishing those meniscus-constrained entities. Small volume; lot of cells. Can I really risk leaving them overnight? Rather, if I’m going to attend my colleague’s forenoon-scheduled workshop, could I really leave them until tomorrow lunchtime? Do I really want to come in, following today’s lengthy, careful preparation, to find them in the midst of acidic death?
This is not the usual experiment I’d set up. However, the opportunity to not spend several hours of each of the next few days – including the weekend – with my forearms in the cell culture hood has been presented me by another refreshingly communicative colleague’s offer of an idea, and materials in the form of coverslips coated with stuff I haven’t considered before (neither of which I wanted to neglectfully waste by risking having the cells peg it). As one of the limiting factors in the problem I repeatedly (unimaginatively) address is that of ‘surface’, it seemed too good to pass up: minimal attention for a few days, then have a look-test. Not much of a hypothesis, but we’ll worry about that later.
So, I about-turned, walked back, and set back to work: added factors to my medium, (commenced scratching this whilst waiting for it to warm), before very gently displacing the surface tension. It won’t work, of course. (Always be negative sceptical) – but you have to be able to say you tried. (Thought: next time, if a repeat is worthwhile… dust off some cloning cylinders.) And it means I’ll have more time at the weekend, perhaps to culture instead my recently burgeoning social life. Spring is here. And I’m still young enough to feel my fanciful thoughts lightly turning…
I have been in that predicament. Happily, I can report that HEK-293 cells (that I work with) are really a lot tougher than what we give them credit for. The same goes for B-cell hybridoma cell lines also. I have stretched their potentials to the limit, and they surprised me. Unfortunately, this is not stuff that gets reported in research papers – so that the evidence remains only anecdotal!
Did I understand that you made more medium and added it into the wells, where you had spotted some cells in a little mound constrained by the size of the droplet? Had they settled and adhered by that point? If so, you won’t have too many problems. And you would have done if you had left them overnight in fifteen microliters… but I can confirm what Kausik says about certain cell lines capacity for rugged survival.
If I’m way off the mark, sorry I misunderstood. But I hope your hope was not misplaced, in any case.
Cheers both. I’m talking primary cells, and not some tough-SoaB (cancer) cell line. But you understood me perfectly, considering I didn’t mention that they had adhered quite nicely.
However, today, they’re not looking quite so healthy, which is affecting the ‘plan’ somewhat.
May be if you sing to them…
Back to seriousness, primary cell cultures are really tough. I have worked with mouse tracheal epithelial cells, as well as lavaged alveolar macrophages, and both of them are a pain to work with. So I can imagine your situation. All the best!
Hey, I wish you two could come to my next lab meeting; I could do with some ‘back-up’.
Sing to them? It’s strange how superstition can kick in, isn’t it? Like if I believe something won’t work, then it might; or if I look too soon, then it hasn’t. Very un-scientific; very Orphean.
Nope, not superstition. Nothing will happen to the cells if you sing to them, but it will make you feel loads better. :)