title: Correlative light and electron microscopy of intermediate stages of meiotic spindle assembly in the early Caenorhabditis elegans embryo.
abstract: This chapter is an update of the previously published book chapter "Correlative Light and Electron Microscopy of Early C. elegans Embryos in Mitosis" (Müller-Reichert, Srayko, Hyman, O'Toole, & McDonald, 2007). Here, we have adapted and improved the protocol for the isolated meiotic embryos, which was necessary to meet the specific challenges a researcher faces while investigating the development of very early Caenorhabditis elegans embryos ex-utero. Due to the incompleteness of the eggshell assembly, the meiotic embryo is very fragile and much more susceptible to changes in the environmental conditions than the mitotic ones. To avoid phototoxicity associated with wide-field UV illumination, we stage the meiotic embryos primarily using transmitted visible light. Throughout the staging and high-pressure freezing, we incubate samples in an isotonic embryo buffer. The ex-utero approach allows precise tracking of the developmental events in isolated meiotic embryos, thus facilitating the comparison of structural features between wild-type and mutant or RNAi-treated samples.
authors.affiliation: Medical Theoretical Center (MTZ), Medical Faculty Carl Gustav Carus, University of Technology Dresden, Fiedlerstraße 42, 01307 Dresden, Germany.
authors.forename: Ina
authors.initials: I
authors.lastname: Woog
authors.forename: Silke
authors.initials: S
authors.lastname: White
authors.forename: Mandy
authors.initials: M
authors.lastname: Büchner
authors.forename: Martin
authors.initials: M
authors.lastname: Srayko
authors.forename: Thomas
authors.initials: T
authors.lastname: Müller-Reichert
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000648:ultrastructure*; D020090:Chromosome Segregation; D002875:Chromosomes / Q000648:ultrastructure; D015925:Cryopreservation; D016616:Cryoultramicrotomy; D055032:Electron Microscope Tomography; D004625:Embryo, Nonmammalian / Q000648:ultrastructure*; D005260:Female; D007091:Image Processing, Computer-Assisted; D008297:Male; D008540:Meiosis; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000648:ultrastructure; D016613:Plastic Embedding; D059705:Polar Bodies / Q000648:ultrastructure