Functionalized contrast agents for multimodality photoacoustic microscopy, optical coherence tomography, and fluorescence microscopy molecular retinal imaging.

Abstract

Near-infrared (NIR) targeting contrast agents have been investigated as great photoabsorbers to improve photoacoustic microscopy (PAM), OCT, and fluorescence imaging contrast for visualization of various diseases. In ophthalmology, a limited number of NIR contrast agents have been approved for clinical use. Recently, gold nanoparticles with different size and shapes have been developed for molecular imaging. This chapter provides the principles of multimodality PAM, OCT, and fluorescence imaging as well as a brief overview of contrast agents for optical imaging. A detailed protocol for the fabrication of discrete colloidal gold nanoparticles (GNPs), synthesis of functionalized RGD-conjugated chain-like GNP (CGNP) clusters labeled with indocyanine green (ICG) fluorescence dye (ICG@CGNP clusters-RGD), and validation of the synthesized nanoparticles to evaluate newly developed blood vessels in the retina, named choroidal neovascularization (CNV), is described. Using RGD peptide, ICG@CGNPs clusters-RGD can bind integrin which is expressed on activated endothelial cells and newly developed CNV. The targeting efficiency of nanoparticles is monitored by multimodality PAM, OCT, and fluorescence imaging longitudinally.