Hi I am using crystal violet assay in microtitre plate for quantifying total biofilm masses formed under different condition and comparing the result. If my OD values are above 3 which is the limit of the Elisa plate reader, i dilute the set of sample and multiply the OD result I get with dilution factor. For example, if i get 2 and dilution factor is 2 i interprete the result as 4. Is it okay?
Even above a figure of OD=2.0 you should be wary of results from the microplate reader. Make sure you adjust the machine to the appropriate volume in your test well.
I routinely diluted my samples so that the OD is below 2.0. However, test samples are compared to control biofilms which are also on the same plate and also to wells with no organisms (negative controls) . Since all the wells are diluted equally, there is no need to adjust OD readings. Biofilm biomass readings should be finally expressed as a proportion of a normal biofilm , with a value of zero being that of the negative control.
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