Daniel:Notebook/HiResChrPaint/2013-9-9
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Fibroblast Adherence Timecourse
The goal of this experiment is to determine the optimal conditions to adhere our fibroblasts on to my poly-L Lysine coated slides for FISH. From previous results it has been suspected that conditions are not right for fibroblast growth. This experiment will be performed on the poly-L Lysine slides as well as glass-bottom culture dishes, which have been ordered from Mattek and are on the way.
Slides
Slide Poly-l-lysine 72/cs Fisher (VWR) #47100
Protocol
1. Cultured cells over the weekend after recovery (PGP1F, Passage 12) 2. Aspirate media and rinse with 5 mL PBS 3. Aspirate PBS and add 1 mL trypsin solution; incubate for several minutes 4. Add 9 mL DMEM to cell culture, mix by gently pipetting 5. Add 100 uL cell suspension to center of glass slides (2x slides) 6. Added 2 mL of cell suspension to 5 different petri dishes containing 8 mL DMEM 7. Took time course photos of slides at 1,2,3,4, and 5 hours 8. Continued 09/10/2013