Troubleshooting

This is likely due to cell culture conditions, which can have a significant impact on micronuclei and EMA-positive values.

Possible Cause:
The actual doubling time of your cell culture is different than expected.

Suggestion:
Establish a growth curve to confirm the doubling time of your culture both after thaw and during routine cell passage. Wait at least one week after thaw before treatment to allow the cells to equilibrate.
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Use your calculated doubling time to determine the correct time to harvest cells (see next question).

Possible Cause:
The media, serum, antibiotics, or other growth factors are not appropriate for your cell line.

Suggestion:
Ensure that you are using cell culture media and supplements that are appropriate for your cell line.

Possible Cause:
The growth environment is not optimal.

Suggestion:
Maintain an incubation temperature of 37 °C, CO₂ levels at 5 %, and humidity near 88 %.

Possible Cause:
The cells are overgrown, which can increase micronuclei and EMA-positive values.

Suggestion:
Ensure that cell cultures do not exceed the following:

Suspension cells: 1.0 x 10⁶ cells/ml
Attachment cells: 80 % confluence

You can calculate % RS using the following formula:

% RS = (Treatment Nuclei:Bead Ratio X 100) / Negative Control Nuclei:Bead Ratio
 

Feel free to send us new questions or example plots to examine. We are always here to help!