Biofilm formation by these isolates was quantitated by measuring OD570 of crystal violet stained wells as described previously (Mohamed, et al., 2004 (link)). Briefly, bacteria that had been grown overnight were diluted 1:100 in tryptic soy broth (TSB) with 0.25% glucose and 200 µl of cells was added onto the wells of polystyrene microtiter plates (Falcon, Franklin Lakes, N.J.). After 24 h of incubation at 37°C, plates were gently washed three-times with phosphate buffered saline (PBS, pH 7.3) and bacteria were fixed with 200 µl of Bouin's fixative for 30 min, washed once with PBS, and stained with 200 µl of crystal violet (1%) for 30 min. After washing and air drying the plates, the biofilm-associated dye was solubilized in 200 µl of ethanol-acetone (80:20, vol/vol) and optical density at 570 nm (OD570) was determined. Each strain was assessed on at least two occasions with 8 replicate wells each time.
Polystyrene microtiter plates
Manufactured by BD
Polystyrene microtiter plates are a type of laboratory equipment used for various applications in scientific research and clinical settings. These plates consist of a grid of small wells made of polystyrene, a common plastic material. The plates are designed to hold small volumes of liquids, such as samples or reagents, and are often used in high-throughput screening, enzyme-linked immunosorbent assays (ELISAs), and other microplate-based assays.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using polystyrene microtiter plates
1
Quantifying Bacterial Biofilm Formation
2
Bacterial Adherence Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Adherence experiments were performed as previously described (84 (link)). Briefly, strains were streaked onto 2YT agar plates and grown overnight at 37°C. Bacteria were scraped from the plates and resuspended in 2YT medium to an OD600 of 0.5, and 100 μl was added into polystyrene microtiter plates (Falcon) and incubated at room temperature for 1 h. Each well then was washed and adhered cells stained with crystal violet. Afterwards, plates were washed and crystal violet dissolved in 70% ethanol at room temperature, and absorbance was measured at 595 nm with a microplate reader (Synergy H1; BioTek). Data analysis was performed to calculate the mean and standard deviation after removal of outliers that were more than one standard deviation from the mean. Data were further normalized to the wild type. Experiments were performed at least three times with up to six technical replicates.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!