Daniel:Notebook/HiResChrPaint/2013-9-12

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Fibroblast Adherence Timecourse (Started 09/09/2013)[edit]

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Timecourse Experiment 09/11/2013[edit]

8. Took another picutre at 24 hours

Results[edit]

Pictures

File:Timecourse 2 091113.png

  • Note: The last time point is 24 hours, not 21 hours

The results don't show it well, but dish 2 did in fact look good, even after 24 hours. The fibroblasts were confluent at that point. The other three looked good for 6 hours, but shriveled between 6 and 24 hours.

Timecourse 3[edit]

Since the dish 2 (Dish, 1:40 dilution trypsin, 1E6 cells/mL, 100 uL DMEM after 3 hours) worked, I'll try that again to see that my results are good. Also preparing cells for passage and freezing using the petri dishes from earlier.

1. Aspirate media
2. Rinse with 5 mL PBS
3. Add 500 uL (dish) or 1 mL (cultures) trypsin LE solution; incubate 5 min
4. Add 9.5 mL (dish) or 9 mL (cultures) DMEM
5. Add 100 uL dish culture solution to center of dish
6. Add 1 mL of culture solution to 9 mL of DMEM (5x) in petri dish
7. Incubate
8. Add 100 uL DMEM to dish after 1 hour of incubation