Improving Laboratory Technique

[Katatonic]

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I've recently gotten the chance to help work on a project in the lab I work at. It's a very interesting and exciting topic, but the actual lab techniques involved are giving me trouble. I'm dealing with living cells and, like so many other lab techniques I'm sure, everything has to be done QUICKLY. The person training me is great, but she's always telling me to work faster work faster work faster. I'm shown something once and then expected to do it myself afterwards which is fine. However, how can I practice these techniques while I'm not at work so that I'm not so slow when time actually counts? The last thing I want to do is do something too slow and kill so many cells that 2 weeks worth of work is useless. All I've been trying is going through the procedures in my head at home, but it's so different than actually working under a tissue culture hood. Thanks for any tips.

 

[GliaGirl]

If your problem is not being able to suck off the old media and feed the cells with new media fast enough, just get an empty plate with no cells on it, and practice sucking water in and out of it under the hood. I'm hard-pressed to think of a way to practice this as home.

Good luck!

 

[Katatonic]

Feeding isn't really my problem, but splitting the cells into a larger number of plates had me a bit stressed haha. Probably only because it was my first time doing something with so many steps involved that wasn't in a lab for a class.

 

[Ariodant]

What's the splitting ratio? I never did more than 1:24, and for those I just mix the trypsinized cells with 260ml of medium (assuming you are using 100mm dishes), and pipet 10ml to each while swirling the bottle constantly.

 

[Lukkie]

first, do some practice problems on paper about splitting cells, ratios, how much media you would need to add to get a 1:8 split or how much trypsin for a 1:3, etc. this is stuff you should be able to do on-the-fly while you're in the hood.

second, if your trainer is trying to have you go so fast during tissue culture, that is a problem. there are so many risks involved, including contaminating the cultures with bacteria or fungi, cross contaminating (mixing up) cell lines, etc. i don't see why they'd start you with timed experiments so quickly when it takes considerable amount of time for anyone to truly learn sterile technique correctly.

no i don't think doing something 'too slow' would kill a lot of cells.

 

[Katatonic]

Thanks for all of the advice, and I'm glad to report back that today, after my second time splitting the cells (by myself w/no supervision) I was much more comfortable, I was going faster but didn't feel rushed, and was much more confident with my work when I put them in the incubator. Each step made much more sense logically to me as well. I suppose practice makes perfect, I was just a bit...frazzled by the new experience a few days ago.

Thanks for the advice about practicing different splitting ratios on paper, I will definitely do that. Luckily, my split today was an easy 1:10. This stuff is so much more interesting that just making gels like usual haha.