PUBLICATION
Labeling cellular structures in vivo using confined primed conversion of photoconvertible fluorescent proteins
- Authors
- Mohr, M.A., Argast, P., Pantazis, P.
- ID
- ZDB-PUB-161105-22
- Date
- 2016
- Source
- Nature Protocols 11: 2419-2431 (Journal)
- Registered Authors
- Pantazis, Periklis (Laki)
- Keywords
- Cell migration, Cellular neuroscience, Fluorescence imaging, Imaging and sensing
- MeSH Terms
-
- Animals
- Light*
- Luminescent Proteins/metabolism*
- Mice
- Microscopy, Confocal/methods*
- Staining and Labeling/methods*
- Zebrafish/embryology
- PubMed
- 27809312 Full text @ Nat. Protoc.
Citation
Mohr, M.A., Argast, P., Pantazis, P. (2016) Labeling cellular structures in vivo using confined primed conversion of photoconvertible fluorescent proteins. Nature Protocols. 11:2419-2431.
Abstract
The application of green-to-red photoconvertible fluorescent proteins (PCFPs) for in vivo studies in complex 3D tissue structures has remained limited because traditional near-UV photoconversion is not confined in the axial dimension, and photomodulation using axially confined, pulsed near-IR (NIR) lasers has proven inefficient. Confined primed conversion is a dual-wavelength continuous-wave (CW) illumination method that is capable of axially confined green-to-red photoconversion. Here we present a protocol to implement this technique with a commercial confocal laser-scanning microscope (CLSM); evaluate its performance on an in vitro setup; and apply primed conversion for in vivo labeling of single cells in developing zebrafish and mouse preimplantation embryos expressing the green-to-red photoconvertible protein Dendra2. The implementation requires a basic understanding of laser-scanning microscopy, and it can be performed within a single day once the required filter cube is manufactured.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping