title: Preparation of Xenopus laevis retinal cryosections for electron microscopy.
abstract: Transmission electron microscopy is the gold standard for examination of photoreceptor outer segment morphology and photoreceptor outer segment abnormalities in transgenic animal models of retinal disease. Small vertebrates such as zebrafish and Xenopus laevis tadpoles have been used to generate retinal disease models and to study outer segment processes such as protein trafficking, and their breeding capabilities facilitate experiments involving large numbers of animals and conditions. However, electron microscopy processing and analysis of these very small eyes can be challenging. Here we present a methodology that facilitates processing of X. laevis tadpole eyes for electron microscopy by introducing an intermediate cryosectioning step. This method reproducibly provides a well-oriented tissue block that can be sectioned with minimal effort by a non-expert, and also allows retroactive analysis of samples collected on slides for light microscopy.
authors.affiliation: University of British Columbia Department of Ophthalmology & Visual Sciences, Canada.
authors.forename: Beatrice M
authors.initials: BM
authors.lastname: Tam
authors.affiliation: University of British Columbia Department of Ophthalmology & Visual Sciences, Canada.
authors.forename: Lee Ling
authors.initials: LL
authors.lastname: Yang
authors.affiliation: University of British Columbia Department of Ophthalmology & Visual Sciences, Canada.
authors.forename: Tami H
authors.initials: TH
authors.lastname: Bogėa
authors.affiliation: University of British Columbia Bioimaging Facility, Canada.
authors.forename: Bradford
authors.initials: B
authors.lastname: Ross
authors.affiliation: University of British Columbia Bioimaging Facility, Canada.
authors.forename: Garnet
authors.initials: G
authors.lastname: Martens
authors.affiliation: University of British Columbia Department of Ophthalmology & Visual Sciences, Canada. Electronic address: olmoritz@mail.ubc.ca.
authors.forename: Orson L
authors.initials: OL
authors.lastname: Moritz
mesh_terms: D000818:Animals; D016616:Cryoultramicrotomy / Q000379:methods*; D016591:Histocytological Preparation Techniques; D046529:Microscopy, Electron, Transmission; D012160:Retina / Q000648:ultrastructure*; D016610:Tissue Embedding; D016707:Tissue Fixation / Q000379:methods; D014982:Xenopus laevis
keywords: Electron microscopy; Outer segment disks; Rods and cones; Ultrastructure; Xenopus laevis