Surgical implantation of an abdominal imaging window for intravital microscopy

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.

[1]  J. van Rheenen,et al.  Brief Report: Intravital Imaging of Cancer Stem Cell Plasticity in Mammary Tumors , 2012, Stem cells.

[2]  Mitsuo Niinomi,et al.  Recent research and development in titanium alloys for biomedical applications and healthcare goods , 2003 .

[3]  K. Svoboda,et al.  Long-term, high-resolution imaging in the mouse neocortex through a chronic cranial window , 2009, Nature Protocols.

[4]  W. Gan,et al.  Two‐photon imaging of dendritic spine development in the mouse cortex , 2008, Developmental neurobiology.

[5]  Rakesh K Jain,et al.  In vivo imaging of extracellular matrix remodeling by tumor-associated fibroblasts , 2009, Nature Methods.

[6]  Jacco van Rheenen,et al.  Intravital imaging of metastatic behavior through a mammary imaging window , 2008, Nature Methods.

[7]  Siqing Shan,et al.  In vivo optical molecular imaging and analysis in mice using dorsal window chamber models applied to hypoxia, vasculature and fluorescent reporters , 2011, Nature Protocols.

[8]  R K Jain,et al.  Vascular permeability and microcirculation of gliomas and mammary carcinomas transplanted in rat and mouse cranial windows. , 1994, Cancer research.

[9]  I. B. Borel Rinkes,et al.  Continuous administration of angiostatin inhibits accelerated growth of colorectal liver metastases after partial hepatectomy. , 2000, Cancer research.

[10]  J. Condeelis,et al.  EGF-induced PIP2 hydrolysis releases and activates cofilin locally in carcinoma cells , 2007, The Journal of cell biology.

[11]  W. Denk,et al.  Two-photon imaging to a depth of 1000 microm in living brains by use of a Ti:Al2O3 regenerative amplifier. , 2003, Optics letters.

[12]  M. Dewhirst,et al.  A novel rodent mammary window of orthotopic breast cancer for intravital microscopy. , 2003, Microvascular research.

[13]  Jochen Herms,et al.  Real-time imaging reveals the single steps of brain metastasis formation , 2010, Nature Medicine.

[14]  Jacco van Rheenen,et al.  Intravital imaging of cell signaling in mice , 2012 .

[15]  Ronald N Germain,et al.  Stromal cell networks regulate lymphocyte entry, migration, and territoriality in lymph nodes. , 2006, Immunity.

[16]  Mikala Egeblad,et al.  Visualizing stromal cell dynamics in different tumor microenvironments by spinning disk confocal microscopy , 2008, Disease Models & Mechanisms.

[17]  Hans Clevers,et al.  Intestinal Crypt Homeostasis Results from Neutral Competition between Symmetrically Dividing Lgr5 Stem Cells , 2010, Cell.

[18]  Bojana Gligorijevic,et al.  Dendra2 Photoswitching through the Mammary Imaging Window , 2009, Journal of visualized experiments : JoVE.

[19]  J. van Rheenen,et al.  Intravital microscopy: new insights into metastasis of tumors , 2011, Journal of Cell Science.

[20]  B. Christ,et al.  The generation of hepatocytes from mesenchymal stem cells and engraftment into murine liver* , 2010, Nature Protocols.

[21]  Thomas A. Cleland,et al.  Chronic in vivo imaging in the mouse spinal cord using an implanted chamber , 2012, Nature Methods.

[22]  K. Messmer,et al.  Dorsal skinfold chamber technique for intravital microscopy in nude mice. , 1993, The American journal of pathology.

[23]  Jacco van Rheenen,et al.  Intravital Microscopy Through an Abdominal Imaging Window Reveals a Pre-Micrometastasis Stage During Liver Metastasis , 2012, Science Translational Medicine.