Skip to Main content Skip to Navigation
Journal articles

Three-Dimensional Imaging of the Vertebral Lymphatic Vasculature and Drainage using iDISCO+ and Light Sheet Fluorescence Microscopy

Abstract : The lymphatic system associated with the central nervous system (CNS) includes the lymphatic vasculature that spins around the brain, the spinal cord, and its associated LNs. The CNS-associated lymphatic system is involved in the drainage of CSF macromolecules and meningeal immune cells toward CNS-draining LNs, thereby regulating waste clearance and immune surveillance within CNS tissues. Presented is a novel approach to obtain three-dimensional (3D) and cellular resolution images of CNS-associated lymphatics while preserving the integrity of their circuits within surrounding tissues. The iDISCO+ protocol is used to immunolabel lymphatic vessels in decalcified and cleared whole mount preparations of the vertebral column that are subsequently imaged with light sheet fluorescence microscopy (LSFM). The technique reveals the 3D structure of the lymphatic network connecting the meningeal and epidural spaces around the spinal cord to extravertebral lymphatic vessels. Provided are 3D images of the drainage circuits of molecular tracers previously injected into either the CSF via the cisterna magna or the thoracolumbar spinal parenchyma. The iDISCO+/LSFM approach brings unprecedented opportunities to explore the structure and function of the CNS-associated lymphatic system in neurovascular biology, neuroimmunology, brain and vertebral cancer, or vertebral bone and joint biology.
Complete list of metadata

Cited literature [24 references]  Display  Hide  Download

https://hal.sorbonne-universite.fr/hal-02934337
Contributor : Hal Sorbonne Université Gestionnaire <>
Submitted on : Wednesday, September 9, 2020 - 11:10:24 AM
Last modification on : Thursday, September 2, 2021 - 9:08:50 AM
Long-term archiving on: : Thursday, December 3, 2020 - 12:45:25 AM

File

document.pdf
Publication funded by an institution

Identifiers

Citation

Laurent Jacob, Jose Brito, Jean-Leon Thomas. Three-Dimensional Imaging of the Vertebral Lymphatic Vasculature and Drainage using iDISCO+ and Light Sheet Fluorescence Microscopy. Journal of visualized experiments : JoVE, JoVE, 2020, pp.e61099 |. ⟨10.3791/61099⟩. ⟨hal-02934337⟩

Share

Metrics

Record views

30

Files downloads

101