Science Is A Bitch
Right when you think you've mastered something and will get results, reality mockingly sinks in, only to taunt you with the notion that you're an ignorant fool who has a long way to go.
No wonder some of the most brilliant scientists I've met are also some of the most humble.
After my first attempt at transfection early in my stay here, Dr. Yu suggested I keep practicing and try a second time to get better results. After repeatedly passaging my 293FT cells and keeping them alive and confluent, I was very proud. Finally, when it came to passaging for the transfection, bam. I did everything more precisely and was extra careful and knew what I was doing, but it didn't matter because my cells were still too few to proceed.
Zhuo assured me not to worry and that the same thing happened to him when he began working with this cell line. Nevertheless I was still forlorn, as I had understood everything and thought my technique was correct. (Plus everytime I mess up I feel like an idiot.) I'm beginning to realize the comprehension of principle is only a minor aspect, and a large part is the refinement and practice of technique. If that's the case, I surely have a long way to go.
To bypass the problem, Zhuo decided to give me a new cell line with which to do tranfection. Instead of the 293FT cells, I used 3T3 fibrolytic stem cells, which are supposedly much less sensitive and adhere to the plate much tighter, making them easier to culture. Today they came into full confluence, so I was culturing and maintaining and seeding like a madwoman. Tomorrow I'm going to transfect, as well as passage the 3T3 GFP cells, which will be compared with the 3T3 control.
Lab stuff seems like a hospital in terms of workload; it really is feast or famine. After all the prep, you just have to wait around for the reactions to take place, and they will take as long as they darn well please.
No wonder some of the most brilliant scientists I've met are also some of the most humble.
After my first attempt at transfection early in my stay here, Dr. Yu suggested I keep practicing and try a second time to get better results. After repeatedly passaging my 293FT cells and keeping them alive and confluent, I was very proud. Finally, when it came to passaging for the transfection, bam. I did everything more precisely and was extra careful and knew what I was doing, but it didn't matter because my cells were still too few to proceed.
Zhuo assured me not to worry and that the same thing happened to him when he began working with this cell line. Nevertheless I was still forlorn, as I had understood everything and thought my technique was correct. (Plus everytime I mess up I feel like an idiot.) I'm beginning to realize the comprehension of principle is only a minor aspect, and a large part is the refinement and practice of technique. If that's the case, I surely have a long way to go.
To bypass the problem, Zhuo decided to give me a new cell line with which to do tranfection. Instead of the 293FT cells, I used 3T3 fibrolytic stem cells, which are supposedly much less sensitive and adhere to the plate much tighter, making them easier to culture. Today they came into full confluence, so I was culturing and maintaining and seeding like a madwoman. Tomorrow I'm going to transfect, as well as passage the 3T3 GFP cells, which will be compared with the 3T3 control.
Lab stuff seems like a hospital in terms of workload; it really is feast or famine. After all the prep, you just have to wait around for the reactions to take place, and they will take as long as they darn well please.
posted by Shannon @ 9:36 PM
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