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Whole mount nuclear fluorescent imaging: Convenient documentation of embryo morphology
Authors:Lisa L Sandell  Hiroshi Kurosaka  Paul A Trainor
Institution:1. Department of Molecular, Cellular and Craniofacial Biology, University of Louisville, Louisville, Kentucky;2. Stowers Institute for Medical Research, Kansas City, Missouri;3. Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, Kansas
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. genesis 2012. © 2012 Wiley Periodicals, Inc.
Keywords:Microscopy  embryology  pseudo‐SEM
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