I performed invasion assay for PC9 cell and I found many way of expressing invaded cells through the matrigel in published papers: 1) simply the number of invaded cells 2) % of invaded cells and 3) invasion index.
Please suggest me which method is the best to explain invaded cells.
Hi,
I think, this mainly depends on your treatment. If your perturbed cells attach like your unperturbed control and also seem to be quite healthy in general, I would just go for the number of invaded cells. This can be controlled by seeding a plate for cell couting or viability assay in parallel to the actual invasion Assay using a dilution of the same cell suspension. In case the results for your perturbed cells are similar to the control, you can go for the number of cells. Otherwise I would normalize to the results from this control plate. You don't want to see differences arising from other factors than invasiveness.
Best,
Cindy
Hi,
I have always counted 5 fields per filter, average them and then average the results of three filters. The data is then presented as number of cells/ field. I think is better than percentage of cells since when a high number of cells are loaded some cells may pile up and not have the same conditions for invasion. In addition, you would have to count all cells in each filter and ,as probably you have experienced, the borders of filters may not reflect the same as the rest of the filter probably due to uneven coating. I think is best to count fields and stay away from filter's edges. If you use a fluorescent system then just express everything as relative invasion, or you can use percentages adjusted to your controls. I hope this helps you,
best,
edgardo
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