Abstract
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.
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Funding
This work was supported by the Central Norway Regional Health Authority “Helse Midt-Norge” [AMS: PhD stipend (90062100), travel grant (90284100); SH: researcher grant (90262100)], and the Norwegian Research Council (SH: 230788/F20).
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All procedures were approved by the Norwegian Animal Research Authorities.
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The authors declare no conflict of interest.
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Sofias, A.M., Åslund, A.K.O., Hagen, N. et al. Simple and Robust Intravital Microscopy Procedures in Hybrid TIE2GFP-BALB/c Transgenic Mice. Mol Imaging Biol 22, 486–493 (2020). https://doi.org/10.1007/s11307-019-01442-2
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DOI: https://doi.org/10.1007/s11307-019-01442-2