12 well polystyrene plate

SYTO9 dye (Invitrogen, Life Technologies) was used to stain nucleic acid of the cells in biofilms and visualized using confocal laser scanning microscopy (CLSM). CLSM was used to confirm the impact of ciprofloxacin (4 and 16 µg/mL), SMJ-5 (32 µg/mL), and a combination of ciprofloxacin (1 µg/mL), and SMJ-5 (32 µg/mL) was used at MBEC₅₀ to visualize the effect. S. aureus SA-1199B biofilm was grown on sterile 18-mm glass coverslips coated with poly-L-lysine placed in a 12-well polystyrene plate (Falcon) (58 (link)). Ciprofloxacin was utilized at 4 and 16 µg/mL; SMJ-5 was used at 32 µg/mL; and a combination of ciprofloxacin at 1 µg/mL and SMJ-5 at 32 µg/mL was used at MBEC₅₀ to visualize the effect. The plates were incubated at 37°C for 24 h. The plates were washed using normal saline to remove planktonic cells. SYTO9 was diluted 1000 times in PBS, poured into the biofilm wells, and left at room temperature for 20–30 min. After incubation, tfhree more saline washes were given, and on an oil immersion lens (×40), images were visualized using Nikon A1 plus Ti confocal microscope with a Nikon A1R scan head. Images were captured using NIS elements software.

SYTO9 dye (Invitrogen, Life Technologies) was used to stain nucleic acid of the cells in biofilms and visualized using confocal laser scanning microscopy (CLSM). CLSM was used to confirm the impact of ciprofloxacin (4 and 16 µg/mL), SMJ-5 (32 µg/mL), and a combination of ciprofloxacin (1 µg/mL), and SMJ-5 (32 µg/mL) was used at MBEC₅₀ to visualize the effect. S. aureus SA-1199B biofilm was grown on sterile 18-mm glass coverslips coated with poly-L-lysine placed in a 12-well polystyrene plate (Falcon) (58 (link)). Ciprofloxacin was utilized at 4 and 16 µg/mL; SMJ-5 was used at 32 µg/mL; and a combination of ciprofloxacin at 1 µg/mL and SMJ-5 at 32 µg/mL was used at MBEC₅₀ to visualize the effect. The plates were incubated at 37°C for 24 h. The plates were washed using normal saline to remove planktonic cells. SYTO9 was diluted 1000 times in PBS, poured into the biofilm wells, and left at room temperature for 20–30 min. After incubation, tfhree more saline washes were given, and on an oil immersion lens (×40), images were visualized using Nikon A1 plus Ti confocal microscope with a Nikon A1R scan head. Images were captured using NIS elements software.