Jacobs:Confocal Imaging of Flow: Difference between revisions
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[[Media:Confocal_imaging_of_cilia_under_fluid_flow.docx]] | [[Media:Confocal_imaging_of_cilia_under_fluid_flow.docx]] | ||
==Procedure== | |||
'''Materials for IMCD Culture''' | |||
* Growth Media: DMEM/F12, 10% FBS, 1% PS, 200ug/ml Geneticin for selection | |||
*Serum Starve Media: change to 0.1% FBS | |||
'''IMCD Culture''' | |||
# Coat the glass bottom of a 35mm glass bottom dish with fibronectin (1:100 in PBS) for 1 hour | |||
# Seed 5k cells in up to 500ul of growth media onto the glass part of a small glass-bottom dish and maintain in culture in for 2 days | |||
Cells will reach ~80% confluency | |||
b. After the cells have adhered (1 hour after seeding) you may add another 1ml of growth media | |||
3. Wash with warm PBS and add 1.5ml of serum starve media and culture for 3 days | |||
Revision as of 07:33, 27 January 2012
Media:Confocal_imaging_of_cilia_under_fluid_flow.docx
Procedure
Materials for IMCD Culture
- Growth Media: DMEM/F12, 10% FBS, 1% PS, 200ug/ml Geneticin for selection
- Serum Starve Media: change to 0.1% FBS
IMCD Culture
- Coat the glass bottom of a 35mm glass bottom dish with fibronectin (1:100 in PBS) for 1 hour
- Seed 5k cells in up to 500ul of growth media onto the glass part of a small glass-bottom dish and maintain in culture in for 2 days
Cells will reach ~80% confluency b. After the cells have adhered (1 hour after seeding) you may add another 1ml of growth media 3. Wash with warm PBS and add 1.5ml of serum starve media and culture for 3 days
