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I observed the basal activity rate daily on MEA. one day, more than three electrodes in the well were spiking regularly in synch but the signal was much weaker a day later. How should I interpret it?
We recommend measuring electrical activity 2~3 hours after changing the media to a fresh one. Nevertheless, various factors can change the signal. When replacing media, cells may be damaged, or the distribution may change as the neurons form clusters.
There is a lot of inter-well variability in the electrical signal. Burst activity can be observed in some wells but not in others. Where does this variability come from, and how can one reduce it?
The cell density is the greatest variable. Plating less cells tends to lead to less variability at the cost of signal amplitude. We suggest plating the cells as evenly as possible.
The Neurosight-S culture is stable over more than 100 days, as long as laminin is supplemented appropriately as detailed in the User Guide. But the period may vary depending on the experimental environment and culture conditions.
When going over the recommended culture period, there may be cell detachment. To prevent this, one method is to add the coating material as a supplement to the cell culture media. For more information, see the user guide.
The electrical activity is very low. Is it possible that culturing the cells longer, over 2 or 3 months, will make the signal increase to acceptable levels? Or should the cells be discarded? After how
We recommend that the cells should be discarded if there is no electrical activity even in 3 weeks. Should this happen, please go over the protocol and reach out to our technical support team at email@example.com.
At what point is it possible to predict whether the electrical activity will increase appropriately over long-term culture?
Normally, you should start to see activity throughout the well 2 weeks after cell plating. If only a few electrodes are active at low spike rates 2 weeks after cell plating, first check whether the cells were plated evenly. The electric acitivty should match what you see visually which means it was a plating issue.
What is the difference between the MEA media(NMS-003, NMS-004) and Neurosight®-S Media (NMS-001, NMS-002)?
The MEA media is optimized to increase the electrical activity of the Neurosight-S within 3 weeks and to the culture conditions(higher density) we recommend on MEA plates. The original media is more adapted to culture at lower densities such as those we recommend in our User Guide for non-electrophysiological assays.
Yes, this is due to the higher plating density required for measuring electrical signals. However, this will not be problematic for signal acquisition(unless the cell cluster moves out of the electrode zone) and is to be expected.
The cell population is stable within one week but electrical signals will take longer to mature (at least 3 weeks).
If there was no problem with cell count or vialbility during cell thawing, there is a high probability that the coating was poor. Please refer to the user guide for coating conditions and methods. Please check with firstname.lastname@example.org for more information.