One of my fellow Rats commented that she liked doing cell culture, because what with the laminar flow hood and everything it felt like she was doing Real Science. I know what she means, so here's another photograph of lab furniture; the cell culture, or laminar flow, hood.
There is an arrangement of fans and filters with the general idea that any nasty stuff inside the hood stays there and does not infect the operator, and everything on the outside stays there and does not infect whatever it is that the operator is working on. Obviously this theory breaks down a little bit because you have to put your hands and flasks of cells, solutions, pipettes and whatnot inside in order to actually do anything. We get around the problems this causes by liberally spraying 70% ethanol over the surface and anything that gets put inside (including hands, which are usually also washed with Hibiscrub first). There is also a reasonably high intensity ultraviolet lamp inside that gets turned on when the hood is not in use, to make life unpleasant for any nasties that do manage to find a way in.
And yes, you do get to feel like a real scientist doing real science, and most of us doing cell culture form an irrational emotional attachment to the cell line(s) we happen to be working with at the moment. We care for, feed and nuture our cells and it can be quite distressing (not just because of the time wasted) when, not if, you go to the incubator one morning and find your precious cultures swarming with bacteria, or fungi, or strange beings from the Planet Claire.
You can imagine, then, that this morning I was someone put out when for the first time in my life I managed to aspirate my trypsinized cell pellet. The cell biologists reading this are cringing now, the rest of you are probably going 'huh?'. Trust me, this is a Bad Thing. No matter, I'll do a Dummies' Guide to Cell Culture one of these days.
I must confess I swore, softly. I went to the incubator, took the second flask that I always keep in case something goes wrong with the first ('real men don't keep backups - they just cry a lot'), did the 70% ethanol routine and started again. I had got as far as adding the trypsin to the cells (in brief, we do this to make the cells come away from the culture plastic surface so that they can be diluted — passaged or 'split' — into new flasks and kept growing. It is also how you get them out of the flask to do an experiment, which was the whole point of me being there this morning), letting the cells sit for a couple of minutes so that they were jumping off the plate into suspension, when the fire alarm rang.
I cursed a little louder this time. Bear in mind that muffing this one would set me back two weeks: I had to think quickly; did I leave them in trypsin (an enzyme that eats protein) and hope I could get back quickly, quench them with media as normal and throw them in the incubator to keep warm, or quench and start the centrifuge and hope they would not chill down too much? I decided to quench and leave them in the incubator, when the alarm stopped.
And started again.
And stopped.
As I still could not smell smoke I decided to push on — Damn the torpedoes! — and hope that I could get the cells safe and warm before it went again.
I need not have worried; a synthetic voice came over the tannoy system stating that it was a false alarm and should be ignored. Then a human voice cut in, saying it was a 'test' (and I swear I heard laughing in the background) and we should 'get back to the bench'. I kid you not.
Shaken, but not stirred, I set up the passage and the experiment, and went on to spray G-418 all over the hood when a syringe filter gave way.
A distinctly sub-optimal start to the day.
Comments
Oh, I just bloody HATED these laminar air flow thingys. First of all, consider that I was doing animal cell culture. To be specific, these were primary cell cultures of murine peritoneal macrophages and mast cells. To the non scientists here, this involved inflating dead mice to hilarious sizes using a saline-like solution (HBSS), jiggling them around and then gently sucking the mixture out with a syringe. They don't require a hell of a lot of care and infections are not a huge problem as we threw the cells away the next day anyhow.
The tubes of HBSS and cells were then spun down in the lab, pellets triturated and so-on until we had clean cells for culture in ... er ... Dulbecco's modified, I think. All that stuff was done in the said evil cabinet.
For me, the pain in the ass factors were three-fold. First up there were four of us doing animal cultures in that lab and only seating space and elbow room for two. Secondly, there was one, single, tree-hugging safety-crazed nutter who was doing human cell culture and needed grade 2 barrier precautions and who also derided anyone using animal cells as they "are not relevant". This coming from someone using U937 human monocytes that have probably has more genetic passages than the Ming Dynasty. Again, for the uninitiated, the human work involves a lab coat that makes you look like a mutant midget chef, shed loads of highly irritant haemosol (that stuff makes great screenwash if you mix it with chloroform, methanol and SDS), and a general air of superior arsiness. The third hassle was the persistence that the cabinet had of delivering Lord-knows-how-many volts of static electricity into my hand whenever I went anywhere near it.
My day-to-day working solution was to eschew my lab coat in favour of a t-shirt, never use anything that wasn't disposable and try my hardest to ignore or counter-irritate Mr U937. On one occasion, that counter-irritation did actually involve the highly-inadvisable method of transferring a tube of multi-drug-resistant malaria to another worker's station by throwing it. I can guarantee that you will never see someone's jaw drop to the floor in such marked horror as we did on that day.
Still, nothing compared to what happened to that dog …
To pass the time between pellet spins, my compatriots and I would devise ever more effective ways of making bombs out of citric acid, bicarbonate, a scintillation vial and a polycarbonate test tube. By the time we had reached what I think was the developmental zenith of this technology, we had a system where a tissue paper wrap of bicarbonate was stuffed into the polycarbonate tube and some concentrated (by that I mean saturated) citric acid was put in the LCS vial. The two components were forced together in such a way that the polycarbonate actually drilled itself into the inner surface of the LCS vial so tightly that you could not actually pull them apart with your hands. If you were careful to keep the assembly upright to keep the reactants apart, you could have an umprimed bomb for all sorts of neat japes.
In terms of power, these things were pretty unbelievable. If one went off in the lab and you happened to be in the same room, then your ears would be left ringing for 5 minutes. If you put one inside an enclosed bin (like a wheelie bin) the effect was closer to a thunderflash going off.
One day, a neighbouring lab, obviously enchanted by the fact that our successes in the field of cell biology was being surpassed by our success in the field of chemical explosives, asked us to show them how it was done. We packed them off with the necessary equipment and waited for the bangs. None came. On asking what had happened, an increasingly amusing story emerged. They had indeed made their bomb and hammered the test tube into the LSC vial with the bottom of a 500ml Gibco bottle. Unfortunately, the boss walked in, so they threw it out the window. The bomb, now primed from its tumble through the air, landed on the grass outside the department where a dog picked it up in its mouth. This was the same guide dog that was walked every day by its blind owner on the grass at the back of the building … I’ll leave it there.
Posted by: Nige | September 20, 2006 10:26 PM
I have to add to my previous comment, that it took certain people in the lab a long time to realise that Dulbecco's modified Eagle medium is not a medium that Mr Dulbecco manufactured from mashed up eagles.
Posted by: Nige | September 21, 2006 12:46 AM
And that, ladies and gentlemen, is why Nigel is now in Marketing.
Posted by: BK | September 21, 2006 09:21 AM
In marketing, you get *paid* for the bangs!
Posted by: Nix | September 22, 2006 06:33 AM