Preparing the CytoView-Z Plate
- Pre-coat the entire well surface of the CytoView-Z plate using an extracellular matrix molecule (ECMM), such as fibronectin.
- Incubate the surface-coated plate in a cell culture incubator at 37°C and 5% CO2 for at least 1 hour.
- Aspirate the surface coating solution from the plate.
- Add 100 µl of complete medium to the plate, and add 8 mL of sterile water to the onplate reservoirs to increase humidity.
- Dock the plate in the Maestro Z and measure the media only (MO) baseline. Transfer the plate to a biosafety cabinet when the baseline is complete.
Culturing Adherent Target Cells and Transferring to CytoView-Z Plate
- Thaw and culture the target cells of interest in accordance with supplier recommendations, passaging as needed.
- Remove flasks of cultured cells from the incubator, aspirate the media, and rinse with warmed phosphate-buffered saline (PBS). Using trypsin, or another cell dissociating agent, detach and collect the cells from the flasks as per reagent recommendations.
- Remove a sample of the cell suspension and count the cells using the Exact FL or a hemocytometer to determine both the cell viability and total number of viable cells.
- Transfer the cell suspension to a 15 ml conical tube and centrifuge the cell suspension to pellet.
- Aspirate the supernatant, being careful not to disturb the cell pellet.
- Dilute the cell suspension in complete medium to the desired working concentration of cells per 100 µl.
Plating Adherent Target Cells onto the CytoView-Z Plate
- Transfer the cell suspension to a reservoir for easy access by a multichannel pipette. Add 100 µl of cell suspension to the 100 µl of media already present in each well.
- Leave the plate seeded with target cells to rest in the biosafety cabinet for 1 hour at room temperature.
- Dock the plate and impedance measurements will begin automatically upon plate engagement.
Dosing Target Cancer Cells with Effector Cells in the CytoView-Z Plate
- Thaw and culture the cells of interest in accordance with supplier recommendations.
- At 24 hrs following target cell seeding, take the flasks of cultured effector cells from the 37°C incubator and mix the suspension gently but thoroughly with a serological pipettor.
- Remove a sample of the cell suspension and count the cells using the Exact FL or a hemocytometer to determine both the viability and total number of viable cells.
- Transfer the cell suspension to a 15 ml conical tube and centrifuge the cell suspension to pellet.
- Aspirate the supernatant, being careful not to disturb the cell pellet.
- Dilute the cell suspension in complete effector cell medium to the desired working concentration of cells per 22.2 µl and add 22.2 µl of effector cell suspension to each well.
- Leave the plate dosed with effector cells to rest in the biosafety cabinet for 30 minutes at room temperature.
- Dock the plate and impedance measurements will automatically resume upon plate engagement.
Required Materials
Consumables
Equipment