High-resolution intravital microscopy through imaging windows has become an indispensable technique for the long-term visualization of dynamic processes in living animals. Easily accessible sites such as the skin, the breast and the skull can be imaged using various different imaging windows; however, long-term imaging studies on cellular processes in abdominal organs are more challenging. These processes include colonization of the liver by metastatic tumor cells and the development of an immune response in the spleen. We have recently developed an abdominal imaging window (AIW) that allows long-term imaging of the liver, the pancreas, the intestine, the kidney and the spleen. Here we describe the detailed protocol for the optimal surgical implantation of the AIW, which takes ∼1 h, and subsequent multiphoton imaging, which takes up to 1 month.
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We thank A. de Graaff and the Hubrecht Imaging Centre for imaging support and T. Faase for help with the replaceable window. This work was supported by a Vidi fellowship (91710330, to J.v.R.) from the Dutch Organization of Scientific Research (NWO), by grants from the Dutch Cancer Society (KWF, HUBR 2009-4621 to J.v.R. and 2009-4367 to E.J.A.S.) and the PON foundation (to E.J.A.S.), and by equipment grants (175.010.2007.00 and 834.11.002) from the NWO.
The authors declare no competing financial interests.
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Ritsma, L., Steller, E., Ellenbroek, S. et al. Surgical implantation of an abdominal imaging window for intravital microscopy. Nat Protoc 8, 583–594 (2013). https://doi.org/10.1038/nprot.2013.026
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