Surgical and intravital microscopy protocol to image Trypanosoma brucei–host interactions in live rodent models

• Published in: STAR protocols Vol. 3; no. 2; p. 101450
• Main Authors: De Niz, Mariana, Figueiredo, Luisa M.
• Format: Journal Article
• Language: English
• Published: Elsevier Inc 17.06.2022; Elsevier
• Subjects: Cell Biology; Microbiology; Microscopy; Model Organisms; Protocol; Model Organisms; Microbiology; Microscopy; Cell Biology
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2022.101450

Intravital microscopy (IVM) involves surgical procedures to expose the internal organs of live anesthetized animals to visualize fluorescently labeled components in situ, in vivo at subcellular resolution. Here, we provide an IVM protocol for time-lapse imaging of dynamic Trypanosoma brucei-host interactions in ten mammalian organs and in systemic circulation. We describe intraperitoneal or intradermal injection of mice with T. brucei. We then detail surgical procedures to prepare ten organs for IVM, followed by imaging of host-T. brucei interactions. For complete details on the use and execution of this protocol, please refer to De Niz et al. (2021). [Display omitted] •An intravital microscopy protocol to study T. brucei in vivo in rodents•Surgical procedures to prepare 10 rodent organs for intravital microscopy•In vivo imaging of host-T. brucei interactions in ten rodent organs and blood vasculature Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Intravital microscopy (IVM) involves surgical procedures to expose the internal organs of live anesthetized animals to visualize fluorescently labeled components in situ, in vivo at subcellular resolution. Here, we provide an IVM protocol for time-lapse imaging of dynamic Trypanosoma brucei-host interactions in ten mammalian organs and in systemic circulation. We describe intraperitoneal or intradermal injection of mice with T. brucei. We then detail surgical procedures to prepare ten organs for IVM, followed by imaging of host-T. brucei interactions.