Leptospirosis Research and Expertise Unit, Institut Pasteur in New Caledonia, Institut Pasteur International Network, Noumea, New Caledonia, France.
Methods Mol Biol. 2020;2134:207-214. doi: 10.1007/978-1-0716-0459-5_19.
Biofilm formation in microtiter plates is certainly the most commonly used method to grow and study biofilm. This simple design is very popular due to its high-throughput screening capacities, low cost, and easy handling. In the protocol described here, we focus on the use of 96-well optically clear, polystyrene flat-bottom plate to study biofilm formation by Leptospira spp. and quantify the biofilm formation by crystal violet (CV) staining. We also describe an alternative method, based on phase contrast image analysis that we believe is more suitable for accurately quantifying biofilm growth by reducing handling of this fragile structure.
在微孔板中形成生物膜无疑是最常用的生长和研究生物膜的方法。由于其高通量筛选能力、低成本和易于处理，这种简单的设计非常受欢迎。在本文描述的方案中，我们专注于使用 96 孔光学透明聚苯乙烯平底板来研究钩端螺旋体属的生物膜形成，并通过结晶紫 (CV) 染色定量生物膜形成。我们还描述了一种基于相差图像分析的替代方法，我们认为该方法通过减少对这种脆弱结构的处理，更适合准确地定量生物膜生长。
