An improved method for three-dimensional reconstruction of protein expression patterns in intact mouse and chicken embryos and organs.

Department of Developmental Biology, Hagedorn Research Institute, Gentofte, Denmark.
Journal of Histochemistry and Cytochemistry (Impact Factor: 2.26). 10/2007; 55(9):925-30. DOI: 10.1369/jhc.7A7226.2007
Source: PubMed

ABSTRACT We have developed a wholemount immunofluorescence protocol for the simultaneous detection of up to three proteins in mouse and chicken embryos. Combined with Murray's clearing reagent (BABB) and microscope objectives with long working ranges and high numerical apertures mounted on a confocal microscope, cellular resolution can be obtained in depths offering the possibility of examining expression patterns in entire organs or embryos. Three-dimensional projections of the optical confocal sections can be computed with computer software allowing rotation around any axis. The protocol is robust and we find that most antibodies working on tissue sections also work with this protocol. This manuscript contains online supplemental material at Please visit this article online to view these materials.

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