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Updated: Sep 9, 2025
Dissection and 2-Photon Imaging of Peripheral Lymph Nodes in Mice
Published on: August 23, 2007
Protocol for rapid 5-plex 3D imaging and single-cell analysis of immune responses in whole murine lymph nodes
Victoire Boulat1, Alessandro Ciccarelli2, Miu Shing Hung3
1Cancer Bioinformatics, Faculty of Life Sciences and Medicine, King's College London, London SE1 3SS, UK; Immunity and Cancer Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
Lymph nodes orchestrate adaptive immune responses, with germinal centers enabling affinity maturation and plasma cell formation. Here, we present a protocol for rapid, high-resolution, multicolor 3D imaging of whole immunized mouse lymph nodes. We describe steps for immunization, lymph node harvesting, fixation, permeabilization, staining, and clearing. We cover image acquisition using a light sheet fluorescence microscope and analysis using the Imaris software. This protocol allows the quantification of germinal center B cells, plasma cells, and follicular T cells at single-cell resolution.
Lymph nodes orchestrate adaptive immune responses, with germinal centers enabling affinity maturation and plasma cell formation. Here, we present a protocol for rapid, high-resolution, multicolor 3D imaging of whole immunized mouse lymph nodes. We describe steps for immunization, lymph node harvesting, fixation, permeabilization, staining, and clearing. We cover image acquisition using a light sheet fluorescence microscope and analysis using the Imaris software. This protocol allows the quantification of germinal center B cells, plasma cells, and follicular T cells at single-cell resolution.
