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Lymph nodes are an important secondary lymphoid organ in the lymphatic system. It is the major site of B cells, T cells and other white blood cells, leading to the function of identifying and fighting infection. To observe the structure of the lymph node, it is necessary to label it with dyes or fluorescent probes. Multi-color labeling are required to label various cell types and structures, which may be difficult under conditions such as in vivo labeling. Stimulated Raman scattering microscopy (SRS) is a burgeoning label-free biological imaging technique that has been used in many areas, such as tumor detection, pharmacokinetics, and neuroscience. Here, we developed a method of label-free imaging of various cell types and structures in lymph nodes with SRS. Collagenous fibers in the lymph node capsule, subcapsular sinus macrophages, B cells, blood vessels and white blood cells were observed in fresh lymph nodes. Based on the visualization of lymph node structure, we prospected its applications in cancer research, aging research and extracellular matrix research.
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