Primo Printing 96 well plate - PEG SVA

Tuesday: 19.02.19.

StockDiluentWorking concentration
5µl 10mM/mL PLL95µl water500µg/ml
5 mg PEG-SVA100µl HEPES (pH 8-8.5)
Printing Protein94ml
  • Add 50µl of Poly-L-Lysine per well
  • Add 300 µl water to peripheral wells
  • Incubate for 30 minutes
  • Rinse 5x with HEPES (100µl per rinse)
  • During rinse step 4/5, dissolve 5 mg PEG-SVA in 50µl HEPES per well), mix with pipette and allow to stand for 1-2 minutes.
  • Add 50µl of PEG-SVA solution to each well
  • Incubate for 1 hour (RT)
  • Rinse 5x with PBS
  • Add 45µl PLPP per well and print

Try to avoid touching the pipette tip off the surface of the glass

Be careful to never allow the surface to dry out

Printing

  • Turn on Tweety, Primo, the Zyla camera and finally the PC.
  • Open Micromanager, then load the primo cfg, and go to Plugins>Leanardo to launch Leanardo.
  • Set the PFS with a fluorescent marker-tagged dish/coverslip with the same thickness as the sample dish.
  • Open the 96 well template, place your pattern and run.
  • After the run is complete, rinse 5x with PBS. Add the protein of interest and incubate at RT for 5'.
  • Rinse 5x with PBS.