Staining Cells on Electrodes using Dapi or NanoOrange

Purpose: To visualize cells on an electrode surface.

Materials needed:

  • 50 mM PIPES buffer
  • DAPI
  • Cells on an electrode

DAPI Procedure

Authors: Heather Jensen & Behzad Rad

  1. Dissolve DAPI to a final concentration of 1 ug/mL in 10mL 50 mM PIPES buffer.
  2. Wash the electrode by dipping it in ~45mL 50 mM PIPES buffer in a 50 mL beaker. Wait for 30 s.
  3. Repeat twice with fresh PIPES buffer.
  4. Immerse the electrode in the 10mL solution of DAPI and incubate for 30s.
  5. Wash the electrode by dipping it in 50 mM PIPES buffer.
  6. Place the electrode on a #1.5 22×60 mm coverslip.
  7. Image with the Zeiss 710.

NanoOrange Procedure

Authors: Sahand Pirbadian & Moh El-Naggar, USC

  1. Deposit 10uL of the culture (OD~0.5) on a glass slide.
  2. Add 0.2 uL of NanoOrange main component.
  3. Mixed the NanoOrange and cells on the glass slide using a pipette.
  4. Cover the slide with a cover-slip and image right away.
  5. You can wait for 10-15 minutes, but in our samples cells were well stained even right after adding the NanoOrange, so there was no need to wait.
  6. Note from Sahand: I hope this is helpful, let me know if you have any questions or if you need the procedure for imaging live cells in a chamber, because we found that to be a little different.

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