A Solid Phase Adherence Assay to Assess Biofilm Formation
Published: March 29, 2024
Abstract
Source: Nickerson, K. P. et al., Bile Salt-induced Biofilm Formation in Enteric Pathogens: Techniques for Identification and Quantification. J. Vis. Exp. (2018)
The video demonstrates a solid-phase adherence assay for assessing biofilm formation. In a multi-well plate, bacteria in bile salt-supplemented media adhere to solid surfaces, forming biofilms, which is confirmed by colorimetry through crystal violet staining and ethanol treatment.
Protocol
1. Preparation of Reagents Bile salts medium: To prepare tryptic soy broth (TSB) containing 0.4% bile salts (weight/volume), resuspend 200 mg of bile salts in 50 mL autoclaved TSB. Filter sterilizes using a 0.22 µm filter. Make fresh medium weekly. NOTES: The bile salts routinely used is a 1:1 mixture of sodium cholate and sodium deoxycholate isolated from ovine and bovine gallbladders. As demonstrated previously, the presence of gluc…
Declarações
The authors have nothing to disclose.
Materials
| Tryptic Soy Broth | Sigma-Aldrich | 22092-500G | |
| Crystal Violet | Sigma | C6158-50 | |
| Bile Salts | Sigma | B8756-100G | |
| LB Agar | Sigma | L7533-1KG | |
| 14 mL culture tubes, 17 x 100 mm, plastic, sterile | Fisher | 14-959-11B | |
| Flat-bottomed 96-well plates (clear) | TPP | 92696 | |
| 96-well plate reader | Spectramax | ||
| 37°C Shaking Incubator | New Brunswick Scientific Excella E25 | ||
| 37°C Plate Incubator | Thermolyne Series 5000 |
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A Solid Phase Adherence Assay to Assess Biofilm Formation. J. Vis. Exp. (Pending Publication), e22064, doi: (2024).