Intravital Imaging of Fluorescent Protein Expression in Mice with a Closed-Skull Traumatic Brain Injury and Cranial Window Using a Two-Photon Microscope
Student Authors
Georgia GunnerAcademic Program
NeuroscienceUMass Chan Affiliations
Morningside Graduate School of Biomedical SciencesNeurobiology
Neurology
Schafer Lab
Document Type
Journal ArticlePublication Date
2023-04-21
Metadata
Show full item recordAbstract
The goal of this protocol is to demonstrate how to longitudinally visualize the expression and localization of a protein of interest within specific cell types of an animal's brain, upon exposure to exogenous stimuli. Here, the administration of a closed-skull traumatic brain injury (TBI) and simultaneous implantation of a cranial window for subsequent longitudinal intravital imaging in mice is shown. Mice are intracranially injected with an adeno-associated virus (AAV) expressing enhanced green fluorescent protein (EGFP) under a neuronal specific promoter. After 2 to 4 weeks, the mice are subjected to a repetitive TBI using a weight drop device over the AAV injection location. Within the same surgical session, the mice are implanted with a metal headpost and then a glass cranial window over the TBI impacting site. The expression and cellular localization of EGFP is examined using a two-photon microscope in the same brain region exposed to trauma over the course of months.Source
Zhong J, Gunner G, Henninger N, Schafer DP, Bosco DA. Intravital Imaging of Fluorescent Protein Expression in Mice with a Closed-Skull Traumatic Brain Injury and Cranial Window Using a Two-Photon Microscope. J Vis Exp. 2023 Apr 21;(194). doi: 10.3791/64701. PMID: 37154548.DOI
10.3791/64701Permanent Link to this Item
http://hdl.handle.net/20.500.14038/52293PubMed ID
37154548ae974a485f413a2113503eed53cd6c53
10.3791/64701