In my experiments using E.coli K-12, the planktonic and biofilm absorbance readings taken at OD600 during the biofilm formation assay are all below 0.1. I have some suggestions of possible sources of error:
1. Too vigorous blotting during the experiment.
2. When immersing plates in water, there may have been residual crystal violet in the water (possible that not all the crystal violet was washed off as a result)
Can anyone help? I'm confused why this has happened.
Mike
Dear Mike, sorry to see that your interesting question has not yet received an expert answer. As an inorganic chemist I'm absolutely not a proven expert in this field. My suggestion would be to check both the "Questions" and "Publications" sections of RG for potentially useful answers and references. For example, I just came across a closely related question which has been posted earlier on RG:
How to conduct a crystal violet assay for quantifying biofilm?
https://www.researchgate.net/post/How_to_conduct_a_crystal_violet_assay_for_quantifying_biofilm
(7 answers)
Yet another closely related and potentially helpful question is the following:
Problem with absorbance in crystal violet assay biofilm?
https://www.researchgate.net/post/Problem_with_absorbance_in_crystal_violet_assay_biofilm
(3 answers)
Here I suggest that you particularly check the literature references provided by Aref Wazwaz. You can also search the "Publications" section of RG e.g. for the term "biofilm crystal violet assay":
https://www.researchgate.net/search.Search.html?type=publication&query=biofilm%20crystal%20violet%20assay
I hope this helps. Good luck with your research and best wishes, Frank Edelmann
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