Indocyanine green fluorescence in second near-infrared (NIR-II) window

PLoS One. 2017 Nov 9;12(11):e0187563. doi: 10.1371/journal.pone.0187563. eCollection 2017.

Abstract

Indocyanine green (ICG), a FDA approved near infrared (NIR) fluorescent agent, is used in the clinic for a variety of applications including lymphangiography, intra-operative lymph node identification, tumor imaging, superficial vascular imaging, and marking ischemic tissues. These applications operate in the so-called "NIR-I" window (700-900 nm). Recently, imaging in the "NIR-II" window (1000-1700 nm) has attracted attention since, at longer wavelengths, photon absorption, and scattering effects by tissue components are reduced, making it possible to image deeper into the underlying tissue. Agents for NIR-II imaging are, however, still in pre-clinical development. In this study, we investigated ICG as a NIR-II dye. The absorbance and NIR-II fluorescence emission of ICG were measured in different media (PBS, plasma and ethanol) for a range of ICG concentrations. In vitro and in vivo testing were performed using a custom-built spectral NIR assembly to facilitate simultaneous imaging in NIR-I and NIR-II window. In vitro studies using ICG were performed using capillary tubes (as a simulation of blood vessels) embedded in Intralipid solution and tissue phantoms to evaluate depth of tissue penetration in NIR-I and NIR-II window. In vivo imaging using ICG was performed in nude mice to evaluate vascular visualization in the hind limb in the NIR-I and II windows. Contrast-to-noise ratios (CNR) were calculated for comparison of image quality in NIR-I and NIR-II window. ICG exhibited significant fluorescence emission in the NIR-II window and this emission (similar to the absorption profile) is substantially affected by the environment of the ICG molecules. In vivo imaging further confirmed the utility of ICG as a fluorescent dye in the NIR-II domain, with the CNR values being ~2 times those in the NIR-I window. The availability of an FDA approved imaging agent could accelerate the clinical translation of NIR-II imaging technology.

MeSH terms

  • Animals
  • Cattle
  • Chickens
  • Indocyanine Green / pharmacology*
  • Optical Imaging / instrumentation*
  • Optical Imaging / methods*
  • Optical Imaging / standards
  • Phantoms, Imaging*
  • Spectroscopy, Near-Infrared / instrumentation
  • Spectroscopy, Near-Infrared / methods
  • Spectroscopy, Near-Infrared / standards

Substances

  • Indocyanine Green

Grants and funding

This work was supported by the Macy Easom Cancer Research Foundation (SAV). There was no additional external funding received for this study. The funder provided support in the form of salaries for authors [ZS, KBG, SAV, AA], but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.