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Media:Confocal_imaging_of_cilia_under_fluid_flow.docx‎


Procedure

Materials for IMCD Culture


IMCD Culture

  1. Coat the glass bottom of a 35mm glass bottom dish with fibronectin (1:100 in PBS) for 1 hour
  2. Seed 3k 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
  1. Wash with warm PBS and add 1.5ml of serum starve media and culture for 3 days

Imaging

  1. Microscope MP off; activate resonance scanner
  2. Use the 100x oil objective
  3. Argon: set to 25 to 30%
  4. Image eGFP: excitation 488/emission 509
  1. Set Frequency: 512 x 512
  1. Use x, y, and z controls to focus on cells and find cilia.

Notes