Parhi Lishay, Shhadeh Amjad, Maalouf Naseem, Alon-Maimon Tamar, Scaiewicz Viviana, Bachrach Gilad
The Institute of Dental Sciences, The Hebrew University-Hadassah School of Dental Medicine, Jerusalem, Israel.
Department of Orthodontics, The Hebrew University-Hadassah School of Dental Medicine, Jerusalem, Israel.
Bio Protoc. 2023 Mar 20;13(6):e4635. doi: 10.21769/BioProtoc.4635.
Over the past decades, the main techniques used to visualize bacteria in tissue have improved but are still mainly based on indirect recognition of bacteria. Both microscopy and molecular recognition are being improved, but most procedures for bacteria detection in tissue involve extensive damage. Here, we describe a method to visualize bacteria in tissue slices from an in vivo model of breast cancer. This method allows examining trafficking and colonization of fluorescein-5-isothiocyanate (FITC)-stained bacteria in various tissues. The protocol provides direct visualization of fusobacterial colonization in breast cancer tissue. Rather than processing the tissue or confirming bacterial colonization by PCR or culture, the tissue is directly imaged using multiphoton microscopy. This direct visualization protocol causes no damage to the tissue; therefore, all structures can be identified. This method can be combined with others to co-visualize bacteria, types of cells, or protein expression in cells.
在过去几十年中，用于在组织中可视化细菌的主要技术有所改进，但仍主要基于对细菌的间接识别。显微镜检查和分子识别技术都在不断改进，但组织中细菌检测的大多数程序都会造成广泛损伤。在这里，我们描述了一种在乳腺癌体内模型的组织切片中可视化细菌的方法。该方法允许检查异硫氰酸荧光素（FITC）染色的细菌在各种组织中的运输和定植情况。该方案提供了乳腺癌组织中梭杆菌定植的直接可视化。该组织不是通过PCR或培养来处理或确认细菌定植，而是使用多光子显微镜直接成像。这种直接可视化方案不会对组织造成损伤；因此，可以识别所有结构。该方法可以与其他方法结合，以共同可视化细菌、细胞类型或细胞中的蛋白质表达。
