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