Neural Organoids on SpheroGuide MEA

Preparing the MEA Plate

1. Place a 5-8 μl drop of 0.1% PEI solution over the recording electrode area of each well in the MEA plate. See Figure 1 on page 2 for appropriate drop placement.

   Tip: Recommended to add 6-8 mL of sterile water to the on-plate reservoirs to increase humidity.

2. Incubate the PEI-coated SpheroGuide plate in a cell culture incubator at 37°C, 5% CO₂ for at least 60 minutes.
3. Rinse PEI from the culture surface with 100 µl of sterile DI water 4 times, then allow the MEA plate to air dry in a biosafety cabinet overnight at room temperature.

Culturing and Plating Neural Organoids

1. Add 200 µl of neural organoid media to each well of a SpheroGuide MEA Plate.
2. Centrifuge at 300 x g for 5 minutes to remove any bubbles.

3. Using a 1mL wide-bore pipette tip, add one neural organoid from culture to each well of the SpheroGuide MEA plate in 100 µl of media. The total well volume will now be 300 µl.

   Tip: Any volume can be used to aspirate the organoid and pipette it into the well. However, it is recommended to not exceed 300 µl per well. It is also recommended to pipette one organoid at a time and to keep the total volume of each well consistent.

4. Check positioning of the organoids by carefully transferring the MEA plate to a microscope. Positioning of the organoid and its contact with the electrodes can also be confirmed via the MEA Viability Module on the Maestro Pro.

   Tip: Some organoid/media compositions may require centrifugation to attach to the bottom of the plate. We recommend starting at 50 x g for 30 seconds and optimizing speed and time as needed.

5. Once the organoids are in place, incubate the SpheroGuide MEA plate in a cell culture incubator at 37°C, 5% CO₂
6. Record activity on the Maestro Pro as needed.
7. For optimal cell health, replace half of the media every 2-3 days. Half-volume (150 µl) media changes are recommended to avoid dislodging the organoid.

Required Materials

Consumables

Equipment