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Surgical implantation of an abdominal imaging window for intravital microscopy

Nature Protocols volume 8, pages 583–594 (2013)Cite this article

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Abstract

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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Figure 1: The specifics of the AIW.
Figure 2: The AIW liver surgery.
Figure 3: Fixation of the mouse in the imaging box.
Figure 4: Tools to retrace areas of interest over multiple days.
Figure 5: Imaging of the liver through the AIW.
Figure 6: Imaging of the spleen through the AIW.
Figure 7: Imaging of the pancreas through the AIW.
Figure 8: Imaging of the small intestine through the AIW.

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Acknowledgements

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.

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Author notes

  1. Laila Ritsma, Ernst J A Steller, Inne H M Borel Rinkes and Jacco van Rheenen: These authors contributed equally to this work.

Authors and Affiliations

  1. Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW)—University Medical Centre Utrecht, Utrecht, The Netherlands

    Laila Ritsma, Saskia I J Ellenbroek & Jacco van Rheenen

  2. Department of Surgery, University Medical Centre Utrecht, Utrecht, The Netherlands

    Ernst J A Steller, Onno Kranenburg & Inne H M Borel Rinkes

  3. Cancer Genomics Netherlands,

    Jacco van Rheenen

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  1. Laila Ritsma
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  2. Ernst J A Steller
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  5. Inne H M Borel Rinkes
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  6. Jacco van Rheenen
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Contributions

L.R. and E.J.A.S. wrote the manuscript, collected the data presented and optimized the surgical procedure and window design. L.R. prepared the figures. J.v.R. conceived and supervised the study and helped write the manuscript. O.K. helped design and supervise the study. I.H.M.B.R. and S.I.J.E. helped to optimize the surgical procedure and window design, respectively, and helped to write the manuscript. All authors had the opportunity to discuss the results and comment on the manuscript.

Corresponding authors

Correspondence to Inne H M Borel Rinkes or Jacco van Rheenen.

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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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