My team is doing DNA extraction from the cells of T. molitor, and for a part of the extraction I need to get the concentration of the cells to 20-30 cells/uL. We’re trying to calculate the concentration by using the OD600 of the cells but we’ve been unable to find any information on that. Is the OD600 of insect cells comparable to mammalian or bacterial cells? If there isn’t an OD600 for insect cells, then is there another way to determine concentration?
Do you have a preparation of separated single cells? If so, simply use a hemocytometer.
Peter
You do not mention what your experiment here is and therefore why you need a measurement and how accurate that measurement must be.
When I have worked with insect cells it is for transfection in reporter assay studies and therefore I need a good level of accuracy to ensure I seed my plates correctly such that they reach the desired level of confluency at the appropriate time. As Peter recommends to you, I use a haemocytometer for this and dilute my suspension appropriately prior to seeding.
If you are forced into doing your work with a spec my suspension is that it will be a low-resolution measurement that will require calibration and hence you will need a haemocytometer for the calibration (preparation of a standard curve). I do feel that even post-calibration it will be low resolution and you would be better served with a haemocytometer.
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