MLNs and spleen were excised and fixed in 4% paraformaldehyde. The tissues were then embedded in paraffin and sectioned at a thickness of 3–5 μm. After dewaxing, dehydration and antigenic reparation, the slices were stained overnight at 4 °C with primary antibodies against CD4, Foxp3, CD80, PD-L1 or CD11c (Servicebio, Wuhan, China) and incubated with HRP-conjugated or FITC-conjugated secondary antibodies (Servicebio, Wuhan, China) for 50 min at room temperature. Cell nuclei were labeled with DAPI for 5 min. The slices were sealed with an anti-fluorescence quenching agent (Servicebio, Wuhan, China) and imaged by fluorescence microscopy (Nikon, Tokyo, Japan).
Fluorescence microscopy
Manufactured by Wuhan Servicebio Technology
Sourced in China
Fluorescence microscopy is a powerful imaging technique that allows for the visualization and analysis of fluorescently labeled samples. The core function of this equipment is to excite fluorescent molecules within a specimen and capture the emitted light, enabling the observation of specific cellular structures, molecules, or processes at a microscopic level.
Automatically generated - may contain errors
2 protocols using fluorescence microscopy
1
Immunophenotyping of Murine Lymphoid Tissues
2
Immunophenotyping of Murine Lymphoid Tissues
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MLNs and spleen were excised and fixed in 4% paraformaldehyde. The tissues were then embedded in paraffin and sectioned at a thickness of 3–5 μm. After dewaxing, dehydration and antigenic reparation, the slices were stained overnight at 4 °C with primary antibodies against CD4, Foxp3, CD80, PD-L1 or CD11c (Servicebio, Wuhan, China) and incubated with HRP-conjugated or FITC-conjugated secondary antibodies (Servicebio, Wuhan, China) for 50 min at room temperature. Cell nuclei were labeled with DAPI for 5 min. The slices were sealed with an anti-fluorescence quenching agent (Servicebio, Wuhan, China) and imaged by fluorescence microscopy (Nikon, Tokyo, Japan).
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