Surface functionalization of barium titanate SHG nanoprobes for in vivo imaging in zebrafish
- Authors
- Culi-Viskota, J., Dempsey, W.P., Fraser, S.E., and Pantazis, P.
- ID
- ZDB-PUB-120822-9
- 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.