Detection of Biofilm Disassembly Through a Dispersion Assay
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 showcases a biofilm dispersion assay using a multi-well plate. Buffer and glucose treatment disassemble the biofilm, releasing bacteria, while bile salts induce stress, retaining bacteria in the protective extracellular polymeric substance. Higher dispersion with buffer and glucose confirms successful biofilm disassembly, estimated by colony forming units measurement.
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 sterilize 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 glucose was required for bile salt-…
Disclosures
The authors have nothing to disclose.
Materials
| Tryptic Soy Broth | Sigma-Aldrich | 22092-500G | |
| Glucose | Sigma | G7021-1KG | |
| 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 | ||
| Flourescent plate reader | Biotek Synergy 2 | ||
| 37°C Shaking Incubator | New Brunswick Scientific Excella E25 | ||
| 37°C Plate Incubator | Thermolyne Series 5000 |
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Cite This Article
Detection of Biofilm Disassembly Through a Dispersion Assay. J. Vis. Exp. (Pending Publication), e22066, doi: (2024).