Jacobs:Confocal Imaging of Flow

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[[Media:Confocal_imaging_of_cilia_under_fluid_flow.docx‎]]
[[Media:Confocal_imaging_of_cilia_under_fluid_flow.docx‎]]
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==Procedure==
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'''Materials for IMCD Culture'''
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* Growth Media: DMEM/F12, 10% FBS, 1% PS, 200ug/ml Geneticin for selection
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*Serum Starve Media: change to 0.1% FBS
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'''IMCD Culture'''
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# Coat the glass bottom of a 35mm glass bottom dish with fibronectin (1:100 in PBS) for 1 hour
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# 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
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Cells will reach ~80% confluency
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b. After the cells have adhered (1 hour after seeding) you may add another 1ml of growth media
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3. Wash with warm PBS and add 1.5ml of serum starve media and culture for 3 days

Revision as of 10:33, 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

  1. Coat the glass bottom of a 35mm glass bottom dish with fibronectin (1:100 in PBS) for 1 hour
  2. 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

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