Today at Applied Biophysics I worked with last week's data I from my co-culturing experiment. Sadly, the data did not turn out as expected. The experiment started out looking promising, however, the cells in every well died out around 15 hours. Our hypothesis as to why this happened? Using an array just after it has been etched is tricky. Arrays that have been recently etched tend to allow more liquid to wick up the sides of the wells. This means that the amount of medium in each well is thinly distributed, and the cells are unable to grow and divide in an environment with insufficient nutrients. Also, this causes the liquid to evaporate faster, thus leaving the remaining medium highly concentrated with saline. Cells cannot survive in such a hypertonic environment- they will shrivel and die due to osmosis.
I treated each of the wells with cysteine, which usually helps with the problem of liquid wicking up the sides of wells, but my array might have been too fresh (last week, I took my array straight from the plasma etcher to the hood). So, this week I repeated our experiment. All conditions remained the same, except I used an array that had been etched a few weeks ago. That should have fixed our problem! Hopefully next week's data goes smoothly!
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