Hi everyone,
I am going to analyse the possible bio-film formation of bacteria fungi exposed to plastic films for about 6 months. What is the procedure to determine bio-film quantity by crystal violet.
The protocol I read was as below:
Take out plastic films from media and rinse it with water to remove unattached microbial strains. Then immerse it in 2 % crystal violet for 15 minute following with rinsing with water and then ethanol or isopropyl.
My question is: after rinsing the plastics it in a clean plate with water and then immersing them in crystal violet, how should I quantify the biofilm? Should I discard the aqueous obtained from washing and immersing the plastic step or I should discard the plastic and focus on the obtained aqueous.
The protocol was not clear and there was not any other protocol for biofilm determination on plastic films, that is why I need your guidance and expertise.
Dear Navid
I have done some work in biofilm formation of bacteria on activated carbon. Following techniques use for detection and quantification of bacteria in biofilms
ESEM (Environmental scanning electomicroscope)
Spread plate technique:
DAPHI Staining
You may use UV-vis spectroscopy to quantify the amount of biofilm. I did some similar work that was published here: Article Antibacterial and Biofilm-Disrupting Coatings from Resin Aci...
You need to discard the water portion to remove free stain. Also, be careful not to remove biofilm from the plastic bag surface when rising with water. Use the ethanol dissolved CV stain for the UV-vis quantification. Good luck!
Dear Mitra,
Many thanks for your answer. What about quantification of fungal biofilm on the plastic? Bacteria would be stainde with cv and de-stained with ethanol so I can check with UV. What if I want it for fungi as well?
Have you done like this before or is there any way?
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