Live imaging of Tribolium castaneum embryonic development using light-sheet-based fluorescence microscopy.
Tribolium castaneum has become an important insect model organism for evolutionary developmental biology, genetics and biotechnology. However, few protocols for live fluorescence imaging of Tribolium have been reported, and little image data is available. Here we provide a protocol for recording the development of Tribolium embryos with light-sheet-based fluorescence microscopy. The protocol can be completed in 4-7 d and provides procedural details for: embryo collection, microscope configuration, embryo preparation and mounting, noninvasive live imaging for up to 120 h along multiple directions, retrieval of the live embryo once imaging is completed, and image data processing, for which exemplary data is provided. Stringent quality control criteria for developmental biology studies are also discussed. Light-sheet-based fluorescence microscopy complements existing toolkits used to study Tribolium development, can be adapted to other insect species, and requires no advanced imaging or sample preparation skills.
Physical Biology/Physikalische Biologie (IZN, FB 15), Buchmann Institute for Molecular Life Sciences (BMLS), Cluster of Excellence Frankfurt-Macromolecular Complexes (CEF-MC), Goethe University-Frankfurt am Main (Campus Riedberg), Frankfurt am Main, Germany.
Frederic
F
Strobl
Physical Biology/Physikalische Biologie (IZN, FB 15), Buchmann Institute for Molecular Life Sciences (BMLS), Cluster of Excellence Frankfurt-Macromolecular Complexes (CEF-MC), Goethe University-Frankfurt am Main (Campus Riedberg), Frankfurt am Main, Germany.
Alexander
A
Schmitz
Physical Biology/Physikalische Biologie (IZN, FB 15), Buchmann Institute for Molecular Life Sciences (BMLS), Cluster of Excellence Frankfurt-Macromolecular Complexes (CEF-MC), Goethe University-Frankfurt am Main (Campus Riedberg), Frankfurt am Main, Germany.
Ernst H K
http://orcid.org/0000-0003-1545-0736
EH
Stelzer
D000818:Animals; D004625:Embryo, Nonmammalian; D004772:Entomology / Q000379:methods*; D008856:Microscopy, Fluorescence; D014231:Tribolium / Q000166:cytology / Q000196:embryology*