Laboratory of Dendritic Cell Biology, Department of Dermatology, University Hospital Erlangen, Friedrich-Alexander-University Erlangen-Nürnberg, Erlangen, Germany.
Department of Physics, Nano-Optics, Sandoghdar Division, Max Planck Institute for the Science of Light, Erlangen, Germany.
Methods Mol Biol. 2021;2350:21-30. doi: 10.1007/978-1-0716-1593-5_2.
Confocal immunofluorescence microscopy is an advanced imaging technique routinely applied in the laboratory and clinics. Histological analyses are performed from tissue material. In general, a single fluorochrome per laser is employed, limiting simultaneous analysis to four antigens in one staining with a conventional 4-laser line microscope. Here, we describe a protocol for combining fluorochromes with the same excitation but different emission properties that allows for the analysis of six different antigens in confocal immunofluorescence microscopy with a conventional 4-laser line microscope. The proposed multiplexed method permits the identification and characterization of complex cell populations in rare tissue material.
共聚焦免疫荧光显微镜是一种常用于实验室和临床的高级成像技术。组织学分析是从组织材料中进行的。一般来说，每个激光使用一种荧光染料，这使得在传统的 4 激光线显微镜中，一次染色最多只能同时分析四种抗原。在这里，我们描述了一种将具有相同激发但不同发射特性的荧光染料结合使用的方案，该方案允许在传统的 4 激光线显微镜下用共聚焦免疫荧光显微镜分析六种不同的抗原。所提出的多重方法允许在稀有组织材料中鉴定和描述复杂的细胞群体。
