An Assay to Study Bile-Salt Induced Biofilm Formation through EPS Matrix Detection

For semi-quantitative detection of EPS, use a multichannel pipette to transfer the culture medium to a clear 96-well plate. Then, use 200 microliters of fixing reagent to fix the black plate for 15 minutes at room temperature. Now, record the OD600 reading by setting the control well as the blank for standardization. Then, record the reading of the culture medium while the adherent population is fixing.

Once the fixing is complete, remove the reagent and discard in the hazardous waste. Then, use 200 microliters of sterile PBS to gently wash the wells twice. After washing, aspirate the PBS. Next, add 150 microliters of 25 micrograms per milliliter of ConA-FITC. Incubate the plate at room temperature for 15 minutes. Post-incubation, gently wash the wells with 200 microliters of PBS. Then, add 150 microliters of PBS to each well, and record the fluorescence at 488 nanometers.