Anatomical 3D Reconstruction of Murine Lymph Nodes for Visualization, Quantitation, and Numerical Simulation

Area of Science:

• Immunology
• Pharmacology
• Biomedical Engineering

Background:

• Lymph nodes (LNs) act as sanctuaries for HIV and cancer due to limited drug penetration.
• Accurate 3D lymph node architecture is crucial for computational modeling of drug transport.
• Existing methods lack the resolution, accessibility, or throughput needed for detailed LN reconstruction.

Purpose of the Study:

• To present a scalable, high-fidelity pipeline for 3D anatomical reconstruction of murine lymph nodes.
• To enable precise mapping of key LN compartments for improved computational modeling.
• To offer an accessible and efficient alternative to current reconstruction methodologies.

Main Methods:

• Integration of optimized vibratome sectioning and multiplexed immunofluorescence staining.
• Confocal microscopy combined with custom automated segmentation algorithms.
• Development of a pipeline for high-fidelity 3D reconstruction of LN architecture.

Main Results:

• Precise reconstruction of lymph node lobules and high endothelial venules with high spatial accuracy.
• Achieved Sørensen-Dice indices >0.93 for lobules and contour-matching scores up to 77% for vasculature.
• Markedly reduced reagent usage (~88%) and labor time (~97%) compared to existing methods.

Conclusions:

• The developed pipeline provides an efficient and broadly accessible approach for high-fidelity 3D LN reconstruction.
• These digital twins of lymph node architecture can support advanced computational simulations.
• Insights from these models can inform therapeutic design for LN-resident diseases and drug delivery.