Simple and Robust Intravital Microscopy Procedures in Hybrid TIE2GFP-BALB/c Transgenic Mice.

Autor: Sofias AM; Department of Circulation and Medical Imaging, Faculty of Medicine and Health Sciences, Norwegian University of Science and Technology (NTNU), Trondheim, Norway. alexandrosofias@outlook.com., Åslund AKO; Department of Physics, Faculty of Natural Sciences, Norwegian University of Science and Technology (NTNU), Trondheim, Norway.; Department of Biotechnology and Nanomedicine, SINTEF Industry, Trondheim, Norway., Hagen N; Comparative Medicine Core Facility, Norwegian University of Science and Technology (NTNU), Trondheim, Norway., Grendstad K; Department of Physics, Faculty of Natural Sciences, Norwegian University of Science and Technology (NTNU), Trondheim, Norway., Hak S; Department of Circulation and Medical Imaging, Faculty of Medicine and Health Sciences, Norwegian University of Science and Technology (NTNU), Trondheim, Norway. sjoerd.hak@ntnu.no.
Jazyk: angličtina
Zdroj: Molecular imaging and biology [Mol Imaging Biol] 2020 Jun; Vol. 22 (3), pp. 486-493.
DOI: 10.1007/s11307-019-01442-2
Abstrakt: Purpose: The endeavor of deciphering intricate phenomena within the field of molecular medicine dictates the necessity to investigate tumor/disease microenvironment real-time on cellular level. We, hereby, design simple and robust intravital microscopy strategies, which can be used to elucidate cellular or molecular interactions in a fluorescent mouse model.
Procedures: We crossbred transgenic TIE2GFP mice with nude BALB/c mice, allowing the breeding of immunocompetent and immunodeficient mouse models expressing green fluorescent protein (GFP) in vascular endothelium. Then, we surgically exposed various tissues of interest to perform intravital microscopy.
Results: By utilizing simple tissue preparation procedures and confocal or two-photon microscopy, we produced high-resolution static snapshots, dynamic sequences, and 3D reconstructions of orthotopically grown mammary tumor, skin inflammation, brain, and muscle. The homogenous detection of GFP expressed by endothelial cells and a combination of fluorescence agents enabled landmarking of tumor microenvironment and precise molecular tagging.
Conclusion: Simple intravital microscopy procedures on TIE2GFP mice allowed a real-time multi-color visualization of tissue microenvironment, underlining that robust microscopy strategies are relatively simple and can be readily available for many tissues of interest.
Databáze: MEDLINE
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