Intravital high-resolution optical imaging of individual vessel response to photodynamic treatment.

Intravital imaging using confocal microscopy facilitates high-resolution studies of cellular and molecular events in vivo. We use this, complemented by Doppler optical coherence tomography (OCT), to assess blood flow in a mouse dorsal skin-fold window chamber model to image the response of individual blood vessels to localized photodynamic therapy (PDT). Specific fluorescent cell markers were used to assess the effect on the vascular endothelial cell lining of the treated vessels. A fluorescently tagged antibody against an endothelial transmembrane glycoprotein (CD31) was used to image endothelial cell integrity in the targeted blood vessel. A cell permeability (viability) indicator, SYTOX Orange, was also used to further assess damage to endothelial cells. A fluorescently labeled anti-CD41 antibody that binds to platelets was used to confirm platelet aggregation in the treated vessel. These optical techniques enable dynamic assessment of responses to PDT in vivo, at both the vascular endothelial cell and whole vessel levels.

[1]  Adrian Mariampillai,et al.  Doppler optical cardiogram gated 2D color flow imaging at 1000 fps and 4D in vivo visualization of embryonic heart at 45 fps on a swept source OCT system. , 2007, Optics express.

[2]  R. G. Statius van Eps,et al.  Platelet Adhesion to Photodynamic Therapy–treated Extracellular Matrix Proteins¶ , 2002 .

[3]  B. Pogue,et al.  Vascular and cellular targeting for photodynamic therapy. , 2006, Critical reviews in eukaryotic gene expression.

[4]  Aleksander Rebane,et al.  Blood-vessel closure using photosensitizers engineered for two-photon excitation , 2008 .

[5]  V. Fingar,et al.  Vascular effects of photodynamic therapy. , 1996, Journal of clinical laser medicine & surgery.

[6]  R. Flower.  Optimizing treatment of choroidal neovascularization feeder vessels associated with age-related macular degeneration. , 2002, American journal of ophthalmology.

[7]  Simon R Cherry,et al.  In vivo molecular and genomic imaging: new challenges for imaging physics. , 2004, Physics in medicine and biology.

[8]  P. Dahm-Kähler,et al.  An intravital microscopy method permitting continuous long-term observations of ovulation in vivo in the rabbit. , 2006, Human reproduction.

[9]  Mamta Khurana,et al.  Simultaneous Two-photon Excitation of Photofrin in Relation to Photodynamic Therapy , 2006, Photochemistry and photobiology.

[10]  B. Furie,et al.  Real-time in vivo imaging of platelets, tissue factor and fibrin during arterial thrombus formation in the mouse , 2002, Nature Medicine.

[11]  M. Kerschensteiner,et al.  Neuroimaging: In vivo imaging of the diseased nervous system , 2006, Nature Reviews Neuroscience.

[12]  C. Garlanda,et al.  Involvement of endothelial PECAM-1/CD31 in angiogenesis. , 1997, The American journal of pathology.

[13]  Xunbin Wei,et al.  Imaging Molecular Expression on Vascular Endothelial Cells by In Vivo Immunofluorescence Microscopy , 2006, Molecular imaging.

[14]  B. Wilson,et al.  Complete blood vessel occlusion in the chick chorioallantoic membrane using two-photon excitation photodynamic therapy: implications for treatment of wet age-related macular degeneration. , 2007, Journal of biomedical optics.

[15]  B. Furie,et al.  In vivo thrombus formation , 2007, Journal of thrombosis and haemostasis : JTH.

[16]  M. Dailey,et al.  Confocal imaging of microglial cell dynamics in hippocampal slice cultures. , 1999, Methods.

[17]  M. Awan,et al.  Review of photodynamic therapy. , 2006, The surgeon : journal of the Royal Colleges of Surgeons of Edinburgh and Ireland.

[18]  D. Broide,et al.  A murine model to study leukocyte rolling and intravascular trafficking in lung microvessels. , 2003, The American journal of pathology.

[19]  Michael L. Dustin,et al.  Dynamic imaging of the immune system: progress, pitfalls and promise , 2006, Nature Reviews Immunology.