Near-Infrared Optical Imaging of Nucleic Acid Nanocarriers In Vivo

Methods Mol Biol. 2019:1943:347-363. doi: 10.1007/978-1-4939-9092-4_23.

Abstract

Noninvasive, real-time optical imaging methods are well suited to follow the in vivo distribution of nucleic acid nanocarriers, their dissociation and the resulting gene expression or inhibition. Indeed, most small animal imaging devices are performing bioluminescence and fluorescence measurements without moving the animal, allowing a simple, rapid, and cost-effective method of investigation of several parameters at a time, in longitudinal experiments that can last for days or weeks.Here we help the reader in choosing adapted near-infrared (NIR) fluorophores or pairs of fluorophores for FRET assays, imaging of reporter genes as well as nanocarriers for in vivo gene and siRNA delivery. In addition, we present the labeling methods of these macromolecules, and of their payload and the protocols to detect them using bioluminescence and NIR fluorescence imaging in mice.

Keywords: In vivo imaging; NIR fluorescence; Nanocarriers; Nucleic acid delivery.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell-Penetrating Peptides / chemistry
  • Female
  • Fluorescence Resonance Energy Transfer / methods*
  • Fluorescent Dyes / chemistry*
  • Gene Transfer Techniques
  • Genes, Reporter / genetics
  • Intravital Microscopy / methods
  • Luminescent Measurements / methods
  • Mice
  • Mice, Nude
  • Microscopy, Fluorescence / methods
  • Nanoparticles / chemistry*
  • Optical Imaging / methods*
  • RNA, Small Interfering / genetics
  • Staining and Labeling / methods

Substances

  • Cell-Penetrating Peptides
  • Fluorescent Dyes
  • RNA, Small Interfering