I want to do plaque assay for which I need host cells in a 6-well culture plate. The protocol requires to plate 293 cells at a density of 70%. How to calculate this density percentage ??
If you would like to be very accurate, you could look what is the surface area of one well of a 6-well plate, and also calculate an approximate area of one cell from photos. Then calculate how many cells you need to seed to make the desired calculate area. But I would not suggest to do it, as after plating you need to wait while the cells attach the plate, they divide and the area could change anyway. Also, is it very important to have exactly 70% of confluency? Usually, from our experience, eg. for cell staining, we try to seed several different concentrations of cell suspension in wells, and then check how do they look like on the next day. If we do not have the desired confluency, we use them one day later. In such way we find the right number of to be seeded empirically, and use it for our next experiments.
In our experiments, 5x105 Vero cells seeded in 6-well plate grow to 70-80% confluency overnight. I suggest to start at concentration of 105 293-cells per well, and then you adjust the concentration till you get the correct density.
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