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…

Divulgations

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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Citer Cet Article
A Solid Phase Adherence Assay to Assess Biofilm Formation. J. Vis. Exp. (Pending Publication), e22064, doi: (2024).

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