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

TY - JOUR

T1 - An improved method for three-dimensional reconstruction of protein expression patterns in intact mouse and chicken embryos and organs

AU - Ahnfelt-Rønne, Jonas

AU - Jørgensen, Mette Christine

AU - Hald, Jacob

AU - Madsen, Ole D

AU - Serup, Palle

AU - Hecksher-Sørensen, Jacob

PY - 2007/9

Y1 - 2007/9

N2 - 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 http://www.jhc.org. Please visit this article online to view these materials.

AB - 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 http://www.jhc.org. Please visit this article online to view these materials.

KW - Animals

KW - Antigens, CD31

KW - Basic Helix-Loop-Helix Transcription Factors

KW - Chick Embryo

KW - Embryo, Mammalian

KW - Embryo, Nonmammalian

KW - Embryonic Development

KW - Fluorescent Antibody Technique

KW - Homeodomain Proteins

KW - Imaging, Three-Dimensional

KW - Mice

KW - Microscopy, Confocal

KW - Nerve Tissue Proteins

KW - Organ Specificity

KW - Trans-Activators

U2 - 10.1369/jhc.7A7226.2007

DO - 10.1369/jhc.7A7226.2007

M3 - Journal article

C2 - 17478445

SN - 0022-1554

VL - 55

SP - 925

EP - 930

JO - Journal of Histochemistry and Cytochemistry

JF - Journal of Histochemistry and Cytochemistry

IS - 9

ER -