title: Whole mount nuclear fluorescent imaging: convenient documentation of embryo morphology.
abstract: Here, we describe a relatively inexpensive and easy method to produce high quality images that reveal fine topological details of vertebrate embryonic structures. The method relies on nuclear staining of whole mount embryos in combination with confocal microscopy or conventional wide field fluorescent microscopy. In cases where confocal microscopy is used in combination with whole mount nuclear staining, the resulting embryo images can rival the clarity and resolution of images produced by scanning electron microscopy (SEM). The fluorescent nuclear staining may be performed with a variety of cell permeable nuclear dyes, enabling the technique to be performed with multiple standard microscope/illumination or confocal/laser systems. The method may be used to document morphology of embryos of a variety of organisms, as well as individual organs and tissues. Nuclear stain imaging imposes minimal impact on embryonic specimens, enabling imaged specimens to be utilized for additional assays.
authors.affiliation: Department of Molecular, Cellular and Craniofacial Biology, University of Louisville, Louisville, KY 40201, USA. Lisa.Sandell@Louisville.edu
authors.forename: Lisa L
authors.initials: LL
authors.lastname: Sandell
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kurosaka
authors.forename: Paul A
authors.initials: PA
authors.lastname: Trainor
mesh_terms: D000818:Animals; D002467:Cell Nucleus; D002642:Chick Embryo; D004622:Embryo, Mammalian / Q000166:cytology*; D004625:Embryo, Nonmammalian / Q000166:cytology*; D005456:Fluorescent Dyes / Q000737:chemistry; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D011894:Rana pipiens; D013194:Staining and Labeling / Q000379:methods*; D015027:Zebrafish
chemical_list: D005456:Fluorescent Dyes