High-Resolution Multi-Photon Imaging of Morphological Structures of Caenorhabditis elegans.
In this protocol, we combine two-photon excitation fluorescence with nonlinear optical measurements to reconstruct the three-dimensional architecture of the pharyngeal region and the muscular system of the anterior and mid-body region of Caenorhabditis elegans (C. elegans). Femto-second laser pulses excite second-harmonic generation (SHG) and third-harmonic generation (THG) signals, which show detailed structural information regarding the organization of myofibrils that are arranged around the central pharynx region. The combination of two-photon excitation with SHG and THG imaging is a very powerful tool to study cell morphology, microarchitecture, and tissue arrangement in C. elegans.
Tissue Morphogenesis, Max Planck Institute for Molecular Biomedicine, and University of Muenster, Muenster, Germany.
Gabriele M
GM
Bixel
Department of Biology, Luther College, Decorah, Iowa.
Stephanie J B
SJ
Fretham
Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, New York.
Michael
M
Aschner
D000818:Animals; D017173:Caenorhabditis elegans / Q000033:anatomy & histology* / Q000648:ultrastructure; D021621:Imaging, Three-Dimensional / Q000379:methods*; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D009132:Muscles / Q000033:anatomy & histology*; D009474:Neurons / Q000648:ultrastructure*; D017711:Nonlinear Dynamics; D010614:Pharynx / Q000033:anatomy & histology*
C. elegans; SHG; THG; label-free imaging; multi-photon microscopy