ZFIN ID: ZDB-PUB-120822-9
Surface functionalization of barium titanate SHG nanoprobes for in vivo imaging in zebrafish
Culi-Viskota, J., Dempsey, W.P., Fraser, S.E., and Pantazis, P.
Date: 2012
Source: Nature Protocols   7(9): 1618-1633 (Journal)
Registered Authors: Dempsey, William, Fraser, Scott E., Pantazis, Periklis (Laki)
Keywords: chemical modification, imaging, model organisms, nanotechnology
MeSH Terms:
  • Animals
  • Barium Compounds/chemistry*
  • Embryo, Nonmammalian/embryology
  • Embryo, Nonmammalian/ultrastructure
  • Microscopy, Confocal
  • Molecular Imaging/methods*
  • Molecular Probes/chemistry*
  • Nanoparticles/chemistry*
  • Nanotechnology/methods*
  • Polyethylene Glycols
  • Titanium/chemistry*
  • Zebrafish/embryology*
PubMed: 22899331 Full text @ Nat. Protoc.

To address the need for a bright, photostable labeling tool that allows long-term in vivo imaging in whole organisms, we recently introduced second harmonic generating (SHG) nanoprobes. Here we present a protocol for the preparation and use of a particular SHG nanoprobe label, barium titanate (BT), for in vivo imaging in living zebrafish embryos. Chemical treatment of the BT nanoparticles results in surface coating with amine-terminal groups, which act as a platform for a variety of chemical modifications for biological applications. Here we describe cross-linking of BT to a biotin-linked moiety using click chemistry methods and coating of BT with nonreactive poly(ethylene glycol) (PEG). We also provide details for injecting PEG-coated SHG nanoprobes into zygote-stage zebrafish embryos, and in vivo imaging of SHG nanoprobes during gastrulation and segmentation. Implementing the PROCEDURE requires a basic understanding of laser-scanning microscopy, experience with handling zebrafish embryos and chemistry laboratory experience. Functionalization of the SHG nanoprobes takes <3 d, whereas zebrafish preparation, injection and imaging setup should take approximately 2–4 h.