Imaging windows for long-term intravital imaging

Intravital microscopy is increasingly used to visualize and quantitate dynamic biological processes at the (sub)cellular level in live animals. By visualizing tissues through imaging windows, individual cells (e.g., cancer, host, or stem cells) can be tracked and studied over a time-span of days to months. Several imaging windows have been developed to access tissues including the brain, superficial fascia, mammary glands, liver, kidney, pancreas, and small intestine among others. Here, we review the development of imaging windows and compare the most commonly used long-term imaging windows for cancer biology: the cranial imaging window, the dorsal skin fold chamber, the mammary imaging window, and the abdominal imaging window. Moreover, we provide technical details, considerations, and trouble-shooting tips on the surgical procedures and microscopy setups for each imaging window and explain different strategies to assure imaging of the same area over multiple imaging sessions. This review aims to be a useful resource for establishing the long-term intravital imaging procedure.

[1]  Stephanie Alexander,et al.  Dynamic imaging of cancer growth and invasion: a modified skin-fold chamber model , 2008, Histochemistry and Cell Biology.

[2]  J. Sandison,et al.  A new method for the microscopic study of living growing tissues by the introduction of a transparent chamber in the rabbit's ear , 1924 .

[3]  R. Dacosta,et al.  Optimization of the dorsal skinfold window chamber model and multi-parametric characterization of tumor-associated vasculature , 2014, Intravital.

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

[5]  Stuart R. Hart,et al.  Unintended perioperative hypothermia. , 2011, The Ochsner journal.

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

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

[8]  R. Glenny,et al.  Stabilized Imaging of Immune Surveillance in the Mouse Lung , 2010, Nature Methods.

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

[10]  C. Hawkey,et al.  Angiogenesis in gastric ulcers: impaired in patients taking non-steroidal anti-inflammatory drugs. , 1995, Gut.

[11]  Becky,et al.  Dexamethasone-induced abolition of the inflammatory response in an experimental glioma model: a flow cytometry study. , 2000, Journal of neurosurgery.

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

[13]  U. Dirnagl,et al.  Electrochemical Failure of the Brain Cortex Is More Deleterious When it Is Accompanied by Low Perfusion , 2013, Stroke.

[14]  C. Heusghem,et al.  Pharmacokinetic Studies of Co‐Trimoxazole in Man After Single and Repeated Doses , 1974, Journal of clinical pharmacology.

[15]  M. Kotiw,et al.  Anti-inflammatory Activity of Hyperimmune Plasma in a Lipopolysaccharide-Mediated Rat Air Pouch Model of Inflammation , 2010, Inflammation.

[16]  Z. Werb,et al.  Monitoring of vital signs for long-term survival of mice under anesthesia. , 2011, Cold Spring Harbor protocols.

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

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

[19]  Ralph Weissleder,et al.  Intravital Imaging , 2011, Cell.

[20]  A. Brunetti,et al.  Mice anesthesia, analgesia, and care, Part I: anesthetic considerations in preclinical research. , 2012, ILAR journal.

[21]  Y. Toiyama,et al.  In Vivo Time-Course Imaging of Tumor Angiogenesis in Colorectal Liver Metastases in the Same Living Mice Using Two-Photon Laser Scanning Microscopy , 2011, Journal of oncology.

[22]  D. Jenkins,et al.  Bioluminescent imaging (BLI) to improve and refine traditional murine models of tumor growth and metastasis , 2004, Clinical & Experimental Metastasis.

[23]  J A Frank,et al.  In vivo three-dimensional MR microscopy of mice with chronic relapsing experimental autoimmune encephalomyelitis after treatment with insulin-like growth factor-I. , 1998, AJNR. American journal of neuroradiology.

[24]  Olav Solgaard,et al.  In vivo brain imaging using a portable 2.9 g two-photon microscope based on a microelectromechanical systems scanning mirror. , 2009, Optics letters.

[25]  Jacco van Rheenen,et al.  Surgical implantation of an abdominal imaging window for intravital microscopy , 2013, Nature Protocols.

[26]  W. Gan,et al.  Choice of cranial window type for in vivo imaging affects dendritic spine turnover in the cortex , 2007, Nature Neuroscience.

[27]  Bernard Choi,et al.  Wide-field functional imaging of blood flow and hemoglobin oxygen saturation in the rodent dorsal window chamber. , 2011, Microvascular research.

[28]  J. van Rheenen,et al.  In vivo imaging and histochemistry are combined in the cryosection labelling and intravital microscopy technique , 2013, Nature Communications.

[29]  M. Schnitzer,et al.  In vivo fluorescence imaging with high-resolution microlenses , 2009, Nature Methods.

[30]  K. Nguyen,et al.  High resolution imaging beyond the acoustic diffraction limit in deep tissue via ultrasound-switchable NIR fluorescence , 2014, Scientific Reports.

[31]  Jun O. Liu,et al.  Effect of nitroxoline on angiogenesis and growth of human bladder cancer. , 2010, Journal of the National Cancer Institute.

[32]  T. Akata.  General Anesthetics and Vascular Smooth Muscle: Direct Actions of General Anesthetics on Cellular Mechanisms Regulating Vascular Tone , 2007, Anesthesiology.

[33]  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.

[34]  Laurie D. Burns,et al.  High-speed, miniaturized fluorescence microscopy in freely moving mice , 2008, Nature Methods.

[35]  Ralph Weissleder,et al.  Advanced Motion Compensation Methods for Intravital Optical Microscopy , 2014, IEEE Journal of Selected Topics in Quantum Electronics.

[36]  R. Shao,et al.  Lidamycin up-regulates the expression of thymidine phosphorylase and enhances the effects of capecitabine on the growth and pulmonary metastases of murine breast carcinoma , 2013, Cancer Chemotherapy and Pharmacology.

[37]  John Condeelis,et al.  Macrophages: Obligate Partners for Tumor Cell Migration, Invasion, and Metastasis , 2006, Cell.

[38]  J. Povlishock,et al.  Therapeutic targeting of the axonal and microvascular change associated with repetitive mild traumatic brain injury. , 2013, Journal of neurotrauma.

[39]  P. Oh,et al.  Co-implanting orthotopic tissue creates stroma microenvironment enhancing growth and angiogenesis of multiple tumors , 2013, F1000Research.

[40]  Ralph Weissleder,et al.  Improved intravital microscopy via synchronization of respiration and holder stabilization , 2012, Journal of biomedical optics.

[41]  David Piwnica-Worms,et al.  On in vivo imaging in cancer. , 2012, Cold Spring Harbor perspectives in biology.

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

[43]  L. Coussens,et al.  The inflammatory tumor microenvironment and its impact on cancer development. , 2006, Contributions to microbiology.

[44]  R. Weissleder,et al.  Automated analysis of clonal cancer cells by intravital imaging , 2013, Intravital.

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

[46]  K. Yamauchi,et al.  Real‐time imaging of single cancer‐cell dynamics of lung metastasis , 2009, Journal of cellular biochemistry.

[47]  J. Segall,et al.  High-resolution multiphoton imaging of tumors in vivo. , 2011, Cold Spring Harbor protocols.

[48]  R. Medema,et al.  Intravital FRET Imaging of Tumor Cell Viability and Mitosis during Chemotherapy , 2013, PloS one.

[49]  H. S. Forbes,et al.  THE CEREBRAL CIRCULATION: I. OBSERVATION AND MEASUREMENT OF PIAL VESSELS , 1928 .

[50]  Dai Fukumura,et al.  In vivo imaging of tumors. , 2010, Cold Spring Harbor protocols.

[51]  Martin Eichner,et al.  Long-Term In Vivo Imaging of β-Amyloid Plaque Appearance and Growth in a Mouse Model of Cerebral β-Amyloidosis , 2011, The Journal of Neuroscience.

[52]  Carlos Portera-Cailliau,et al.  A craniotomy surgery procedure for chronic brain imaging. , 2008, Journal of visualized experiments : JoVE.

[53]  Ralph Weissleder,et al.  Automated motion artifact removal for intravital microscopy, without a priori information , 2014, Scientific Reports.

[54]  A. Bauer,et al.  Antibiotic susceptibility testing by a standardized single disk method. , 1966, American journal of clinical pathology.

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

[56]  M. Dewhirst,et al.  High-resolution in vivo imaging of fluorescent proteins using window chamber models. , 2012, Methods in molecular biology.

[57]  Hans Clevers,et al.  Intestinal crypt homeostasis revealed at single stem cell level by in vivo live-imaging , 2014, Nature.

[58]  David Kleinfeld,et al.  A polished and reinforced thinned-skull window for long-term imaging of the mouse brain. , 2012, Journal of visualized experiments : JoVE.

[59]  Chung-Hyun Cho,et al.  COMP-angiopoietin-1 promotes wound healing through enhanced angiogenesis, lymphangiogenesis, and blood flow in a diabetic mouse model. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[60]  P. Friedl,et al.  Intravital third harmonic generation microscopy of collective melanoma cell invasion , 2012, Intravital.

[61]  Paul Salama,et al.  Two-photon imaging within the murine thorax without respiratory and cardiac motion artifact. , 2011, The American journal of pathology.

[62]  Deborah S. Barkauskas,et al.  Comparison of intravital thinned skull and cranial window approaches to study CNS immunobiology in the mouse cortex , 2014, Intravital.

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

[64]  Hiroki Iida,et al.  Isoflurane and Sevoflurane Induce Vasodilation of Cerebral Vessels via ATP‐sensitive K+ Channel Activation , 1998, Anesthesiology.

[65]  Y. Toiyama,et al.  In vivo real-time imaging of chemotherapy response on the liver metastatic tumor microenvironment using multiphoton microscopy. , 2012, Oncology reports.

[66]  N. Kasthuri,et al.  Long-term dendritic spine stability in the adult cortex , 2002, Nature.

[67]  S. Gambhir,et al.  Nondestructive, serial in vivo imaging of a tissue-flap using a tissue adhesion barrier , 2012 .

[68]  Yaniv Ziv,et al.  Time-lapse imaging of disease progression in deep brain areas using fluorescence microendoscopy , 2011, Nature Medicine.

[69]  Chris I. De Zeeuw,et al.  Elimination of Inhibitory Synapses Is a Major Component of Adult Ocular Dominance Plasticity , 2012, Neuron.

[70]  R. Weissleder,et al.  Real-time in vivo imaging of the beating mouse heart at microscopic resolution , 2012, Nature Communications.

[71]  D. Resnik,et al.  The agony of agonal respiration: is the last gasp necessary? , 2002, Journal of medical ethics.

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

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

[74]  M. Dewhirst,et al.  Radiation activates HIF-1 to regulate vascular radiosensitivity in tumors: role of reoxygenation, free radicals, and stress granules. , 2004, Cancer cell.

[75]  K. Nguyen,et al.  High-resolution imaging in a deep turbid medium based on an ultrasound-switchable fluorescence technique. , 2012, Applied physics letters.

[76]  K. Ohsaka,et al.  Comparison of newly developed inhalation anesthesia system and intraperitoneal anesthesia on the hemodynamic state in mice. , 2007, Biological & pharmaceutical bulletin.

[77]  S. Ramaswamy,et al.  Systematic identification of genomic markers of drug sensitivity in cancer cells , 2012, Nature.

[78]  A. Holtmaat,et al.  Dendritic Spine Instability Leads to Progressive Neocortical Spine Loss in a Mouse Model of Huntington's Disease , 2013, The Journal of Neuroscience.

[79]  D. Schwenke,et al.  Comparison of the depressive effects of four anesthetic regimens on ventilatory and cardiovascular variables in the guinea pig. , 2004, Comparative medicine.

[80]  Franck Debarbieux,et al.  Long‐term in vivo imaging of normal and pathological mouse spinal cord with subcellular resolution using implanted glass windows , 2012, The Journal of physiology.

[81]  D. Kobat,et al.  In vivo two-photon microscopy to 1.6-mm depth in mouse cortex. , 2011, Journal of biomedical optics.

[82]  Jacco van Rheenen,et al.  Imaging hallmarks of cancer in living mice , 2014, Nature Reviews Cancer.

[83]  William A Mohler,et al.  Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues. , 2002, Biophysical journal.

[84]  Michael Leunig,et al.  The dorsal skinfold chamber: studying angiogenesis by intravital microscopy. , 2009, Methods in molecular biology.

[85]  Mikala Egeblad,et al.  Caught in the act: revealing the metastatic process by live imaging , 2013, Disease Models & Mechanisms.

[86]  S. Kaplan,et al.  Pharmacokinetic profile of trimethoprim-sulfamethoxazole in man. , 1973, The Journal of infectious diseases.

[87]  Y. Castier,et al.  Endothelin receptor antagonism prevents hypoxia-induced mortality and morbidity in a mouse model of sickle-cell disease. , 2008, The Journal of clinical investigation.

[88]  Jochen Herms,et al.  Imaging glioma cell invasion in vivo reveals mechanisms of dissemination and peritumoral angiogenesis , 2009, Glia.

[89]  Michael Thomaschewski,et al.  RGB marking facilitates multicolor clonal cell tracking , 2011, Nature Medicine.

[90]  G. H. Algire.  An Adaptation of the Transparent-Chamber Technique to the Mouse , 1943 .

[91]  J. Rheenen,et al.  Real-time intravital imaging of cancer models , 2011, Clinical & translational oncology : official publication of the Federation of Spanish Oncology Societies and of the National Cancer Institute of Mexico.

[92]  C. Porro,et al.  Ketamine Effects on Local Cerebral Blood Flow and Metabolism in the Rat , 1987, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[93]  A. Mizrahi,et al.  Long-Term Imaging Reveals Dynamic Changes in the Neuronal Composition of the Glomerular Layer , 2011, The Journal of Neuroscience.

[94]  Frank Winkler,et al.  Insights into cell-to-cell and cell-to-blood-vessel communications in the brain: in vivo multiphoton microscopy , 2013, Cell and Tissue Research.

[95]  P. Friedl.  Dynamic imaging of cellular interactions with extracellular matrix , 2004, Histochemistry and Cell Biology.

[96]  M. Menger,et al.  In vivo analysis of microvascular reperfusion injury in striated muscle and skin , 1994, Microsurgery.

[97]  C. Portera-Cailliau,et al.  A method for 2-photon imaging of blood flow in the neocortex through a cranial window. , 2008, Journal of visualized experiments : JoVE.

[98]  S. Houten,et al.  Optimizing anesthetic regimen for surgery in mice through minimization of hemodynamic, metabolic, and inflammatory perturbations , 2014, Experimental biology and medicine.

[99]  Z. Werb,et al.  Preparation of mice for long-term intravital imaging of the mammary gland. , 2011, Cold Spring Harbor protocols.

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