4.4. Quantitative Fluorescence-Based Biofilm Measurement (Resazurin Assay)
This protocol is extracted from research article:
Inhibition of Biofilm Formation by the Synergistic Action of EGCG-S and Antibiotics
Antibiotics (Basel), Jan 21, 2021; DOI: 10.3390/antibiotics10020102

Fresh log-phase bacterial cultures were incubated at 37 °C for 4 days in 96-well plates. After the media was aspirated, the biofilm was rinsed with 1 × PBS and stained with a 200 μM Resazurin solution. After overnight incubation in the dark at 4 °C, fluorescence was measured at excitation and an emission wavelength of 560 nm and 590 nm respectively using a microplate reader (Infinite 200 PRO, Tecan, Männedorf, Switzerland) adjusted to the fluorescent mode [47]. The Relative Light Units (RLU) were measured, and the percentage (%) of inhibition was calculated for each treatment according to the following formula.

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