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...

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Bibliographic Details
Published inCurrent protocols in toxicology Vol. 64; p. 11.19.1
Main Authors Bixel, Gabriele M, Fretham, Stephanie J B, Aschner, Michael
Format Journal Article
LanguageEnglish
Published United States 01.05.2015
Subjects
Animals
Caenorhabditis elegans - anatomy & histology
Caenorhabditis elegans - ultrastructure
Imaging, Three-Dimensional - methods
Microscopy, Fluorescence, Multiphoton - methods
Muscles - anatomy & histology
Neurons - ultrastructure
Nonlinear Dynamics
Pharynx - anatomy & histology
label-free imaging
C. elegans
THG
SHG
multi-photon microscopy
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Summary: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.
ISSN:1934-9262
DOI:10.1002/0471140856.tx1119s64