Imaging Glioma Progression by Intravital Microscopy
We describe here a method for generating mouse orthotopic gliomas in order to follow their progression over time by multi-photon laser scanning microscopy. After craniotomy of the parietal bone, glioma cells are implanted in the brain cortex and a glass window is cemented atop, allowing chronical imaging of the tumor. The expression of different fluorescent proteins in tumor cells and in specific cell types of a number of currently available transgenic mouse strains allows obtaining multicolor 3D images of the tumor over time. This technique is suitable both to evaluate the effect of pharmacological treatments and to unravel basic mechanisms of tumor-host interactions.
Key wordsIntravital imaging Multi-photon Laser scanning microscopy Tumor model Glioma
The development of the technique here described was supported by the Belgian Cancer Foundation (Stichting Tegen Kanker, grant 2012‐181) and a Hercules type 2 grant (Herculesstichting: AKUL11033). We thank Dr. Till Acker (Institute of Neuropathology, University of Giessen, Germany) and Dr. Thomas N. Seyfried (Biology department, Boston College, USA) for the gift of Glioma261 and CT2A cells, respectively.
- 7.Ricard C, Stanchi F, Rougon G, Debarbieux F (2014) An orthotopic glioblastoma mouse model maintaining brain parenchymal physical constraints and suitable for intravital two-photon microscopy. J Vis Exp. https://doi.org/10.3791/51108
- 10.Mostany R, Portera-Cailliau C (2008) A method for 2-photon imaging of blood flow in the neocortex through a cranial window. J Vis Exp. https://doi.org/10.3791/678