Microscopy & ImagingJoVE (Open Access)Citable · DOI
Intravital Imaging of the Mouse Thymus using 2-Photon Microscopy
DOI: 10.3791/3504-v
What you'll learn
✓Perform bone marrow transplantation and thymic reconstitution in mice
✓Execute surgical exposure of transplanted thymus tissue
✓Acquire intravital two-photon microscopy images of thymic T cell development
✓Identify and visualize T cell developmental niches within thymic tissue
Protocol
We have developed novel laboratory tools and protocols for intravital imaging acquisition of the thymus. Our technique should help in the identification of “niches” within the thymus where T cell development occurs.
Difficulty
advanced
Total time
~4–6 weeks (includes irradiation, bone marrow transfer, transplantation healing, and imaging surgery per mouse)
Overview of novel protocols for acquiring intravital images of the mouse thymus and identifying T cell developmental niches using two-photon microscopy.
▶ 00:05
2
Perform irradiation and bone marrow transfer
Irradiate recipient mice and transplant donor bone marrow to establish hematopoietic chimerism necessary for thymic reconstitution.
▶ 01:15
3
Conduct thymic tissue transplantation surgery
Transplant thymic tissue into prepared recipient mice to establish functional thymic tissue for intravital imaging.
▶ 03:16
4
Surgically expose transplanted thymus tissue
Perform surgical preparation to access and expose the transplanted thymus for imaging while maintaining tissue viability.
▶ 04:58
5
Acquire two-photon intravital microscopy images
Perform two-photon microscopy image acquisition of exposed thymic tissue to visualize T cell development and identify developmental niches.