Mhade Shreeya, Kaushik Karishma S
Department of Biotechnology, Savitribai Phule Pune University, Pune 411007, India.
ACS Omega. 2023 May 26;8(23):20163-20177. doi: 10.1021/acsomega.2c07255. eCollection 2023 Jun 13.
Confocal laser-scanning microscopy (CLSM) is the bedrock of the microscopic visualization of biofilms. Previous applications of CLSM in biofilm studies have largely focused on observations of bacterial or fungal elements of biofilms, often seen as aggregates or mats of cells. However, the field of biofilm research is moving beyond qualitative observations alone, toward the quantitative analysis of the structural and functional features of biofilms, across clinical, environmental, and laboratory conditions. In recent times, several image analysis programs have been developed to extract and quantify biofilm properties from confocal micrographs. These tools not only vary in their scope and relevance to the specific biofilm features under study but also with respect to the user interface, compatibility with operating systems, and raw image requirements. Understanding these considerations is important when selecting tools for quantitative biofilm analysis, including at the initial experimental stages of image acquisition. In this review, we provide an overview of image analysis programs for confocal micrographs of biofilms, with a focus on tool selection and image acquisition parameters that are relevant for experimental researchers to ensure reliability and compatibility with downstream image processing.
共聚焦激光扫描显微镜（CLSM）是生物膜微观可视化的基础。CLSM先前在生物膜研究中的应用主要集中于观察生物膜中的细菌或真菌成分，这些成分通常被视为细胞聚集体或菌毯。然而，生物膜研究领域正从单纯的定性观察，转向对临床、环境和实验室条件下生物膜结构和功能特征的定量分析。近年来，已经开发了几个图像分析程序，用于从共聚焦显微照片中提取和量化生物膜特性。这些工具不仅在其范围和与所研究的特定生物膜特征的相关性方面有所不同，而且在用户界面、与操作系统的兼容性以及原始图像要求方面也存在差异。在为生物膜定量分析选择工具时，包括在图像采集的初始实验阶段，了解这些注意事项很重要。在这篇综述中，我们概述了用于生物膜共聚焦显微照片的图像分析程序，重点关注与实验研究人员相关的工具选择和图像采集参数，以确保可靠性和与下游图像处理的兼容性。
