Abstract：
Given the significant impact of biofilms on human health and material corrosion, research in this field urgently needs more accessible techniques to facilitate the testing of new control agents and general understanding of biofilm biology. Microtiter plates offer a convenient format for standardized evaluations, including high-throughput assays of alternative treatments and molecular modulators. This study introduces a novel Biofilm Analysis Software (BAS) for quantifying biofilms from microtiter plate images. We focused on early biofilm growth stages and compared BAS quantification to common techniques: direct turbidity measurement, intrinsic fluorescence detection linked to pyoverdine production, and standard crystal violet staining which enables image analysis and optical density measurement. We also assessed their sensitivity for detecting subtle growth effects caused by cyclic AMP and gentamicin. Our results show that BAS image analysis is at least as sensitive as the standard method of spectrophotometrically quantifying the crystal violet retained by biofilms. Furthermore, we demonstrated that bacteria adhered after short incubations (from 10 min to 4 h), isolated from planktonic populations by a simple rinse, can be monitored until their growth is detectable by intrinsic fluorescence, BAS analysis, or resolubilized crystal violet. These procedures are widely accessible for many laboratories, including those with limited resources, as they do not require a spectrophotometer or other specialized equipment.
摘要：
鉴于生物膜对人体健康和材料腐蚀的重大影响，该领域的研究迫切需要更容易获得的技术，以促进新控制剂的测试和对生物膜生物学的一般理解。微量滴定板提供了一个方便的格式的标准化评价，包括替代治疗和分子调节剂的高通量测定。本研究介绍了一种新颖的生物膜分析软件（BAS），用于从微量滴定板图像定量生物膜。我们专注于早期生物膜生长阶段，并将BAS定量与常见技术进行了比较：直接浊度测量，与pyoverdine生产相关的固有荧光检测，和标准结晶紫染色，使图像分析和光密度测量。我们还评估了它们对检测由环状AMP和庆大霉素引起的细微生长效应的敏感性。我们的结果表明，BAS图像分析至少与分光光度法定量生物膜保留的结晶紫的标准方法一样灵敏。此外，我们证明了细菌在短暂孵育(从10分钟到4小时)后粘附，通过简单的冲洗从浮游种群中分离出来，可以监测，直到它们的生长被内在荧光检测到，BAS分析,或重新溶解的结晶紫。许多实验室可以广泛使用这些程序，包括那些资源有限的人，因为它们不需要分光光度计或其他专用设备。
