Daniel:Notebook/HiResChrPaint/2013-9-10

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Revision as of 00:39, 13 September 2013 by >Djacobse (→‎Results)
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Fibroblast Adherence Timecourse

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Slides (From 09/09/2013)

9. Took photo at time point 21 hours

Even after 22 hours, the slides appeared to have few cells, and the cells present were not well attached. It is my theory that 1 of two things are happening: (1) The poly-L lysine slides are not providing strong support for the cells and they are not adhering; or (2) the cells are not getting enough nutrients. The second hypothesis is easier to test, so I am going to try that. If I can recover these cells with media, that will, I think, prove my point. Otherwise, I'll have to repeat the experiment. If I do that, I'd recommend:

  • Trypsin (1 mL)
  • Add DMEM to get to ~1 000 000 cells/mL, then add 100 uL to center of the slide
  • Wait 1-3 hours for basic adherence, then add 3 mL DMEM to promote more growth.


10. Added 3 mL DMEM to each culture dish (22 hours)

Results

File:Timecourse FinalComparison 090913.png

There was no improvement in the appearance of the slides at 3 hours post-media introduction, nor 6 hours post-media introduction. I didn't even bother to take pictures, the cells just looked dead. I think that they grew too long with only 100 uL media.