In Vitro Biofilm Synthesis by Staphylococcus aureus

Published: September 29, 2023

Abstract

Source: Rana, P. S. et al., Standardized In vitro Assays to Visualize and Quantify Interactions between Human Neutrophils and Staphylococcus aureus Biofilms. J. Vis. Exp. (2022)

In this video, we demonstrate the preparation of in vitro biofilm by S. aureus in a poly-L-Lysine-coated multi-well plate. The biofilm-formed plate can be used for downstream experiments.

Protocol

1. Preparation of in vitro biofilm Obtain isolated colonies of S. aureus from a cryopreserved stock using a streak-plate technique on a nutrient-rich agar plate, such as Tryptic Soy Agar (see Table of Materials). Coat individual wells of a 96-well plate with 100 µL of 0.001% (v/v) poly-L-Lysine (PLL) diluted in sterile H2O and incubate at room temperature for 30 min. Aseptically, aspirate the PLL solution using a vacuum-assisted asp…

Divulgations

The authors have nothing to disclose.

Materials

BD Bacto Tryptic Soy Broth BD DF0370-07-5 Combine with 1.5% agar to make Tryptic Soy Agar
Clear bottom 96-well flat bottom polystyrene plates Costar 3370
Hanks' balanced salt solution (HBSS) 1x Corning cellgro 21-022-CV Without calcium, magnesium, and phenol red
Poly-L-lysine solution Sigma P4707-50ML
White opaque 96-well plates Falcon 353296 Tissue culture treated and flat-bottom plate
Minimal essential media (MEM) Alpha 1x Gibco 41061-029

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In Vitro Biofilm Synthesis by Staphylococcus aureus. J. Vis. Exp. (Pending Publication), e21619, doi: (2023).

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