Daniel:Notebook/HiResChrPaint/2013-9-9

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

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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

Results

Slide 1

File:Slide 1 Timecourse 090913.png

Slide 2

File:Slide 2 Timecourse 090913.png

Neither slide showed any real growth during the experiment. I'm leaving them overnight to keep checking.