Priming BioFlux plates

Procedures for 48-well low shear, 48-well high shear, 24-well, and 6-well plates

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This procedure aims to minimize the formation of air bubble in the channels of the BioFlux plates during priming. All materials and reagents should be equilibrated to the same temperature before using (i.e. i running at 37°C, plate, media, and reagents should be at 37°C; perform priming also at 37°C)

You may apply this prime protocol as a coating protocol. Replace PBS with the coating
solution. No need to prime and then coat the plate.

Prime low-shear (0-20dyn) bcxx 48-well plate

  1. Load at least 40uL of PBS to the outlet well.
  2. Place the interface cover on the plate and ensure a snug/tight seal
  3. Perfuse from outlet to inlet (if using autorun, set as reverse) at 20dyn/cm2 for 60 seconds. 
  4. Direction of flow

Prime high-shear (0-200dyn) caxx 48-well plate

  1. Load at least 40uL of PBS to the inlet well.
  2. Place the interface cover on the plate and ensure a snug/tight seal
  3. Perfuse from inlet to outlet (if using autorun, set as forward) at 200dyn/cm2 for 4 minutes.
  4. Direction of flow

Prime low-shear (0-20dyn) aaxx 24-well plate

  1. Load at least 100uL of PBS to the outlet well.
  2. Place the interface cover on the plate and ensure a snug/tight seal
  3. Perfuse from outlet to inlet (if using autorun, set as reverse) at 20dyn/cm2 for 60 seconds.
  4. Direction of flow

Prime (0-60dyn) abxx 6-well plate

  1. Load at least 200uL of PBS to the outlet well.
  2. Place the interface cover on the plate and ensure a snug/tight seal
  3. Perfuse from outlet to inlet (if using autorun, set as reverse) at 60dyn/cm2 for 60 seconds.
  4. Direction of flow