Cite as: Cold Spring Harb. Protoc.; 2009; doi:10.1101/pdb.prot5335

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Collection, Handling, Fixation, and Processing of Tammar Wallaby (Macropus eugenii) Embryos

Danielle Hickford, Stephen Frankenberg, and Marilyn B. Renfree1

Department of Zoology, The University of Melbourne, Victoria 3010, Australia

1Corresponding author (m.renfree{at}unimelb.edu.au).


INTRODUCTION

This protocol describes methods for collecting and handling tammar wallaby (Macropus eugenii) conceptuses. Zygotes, cleavage stages, diapausing blastocysts, and unexpanded blastocysts are ~250 µm in diameter and are collected by flushing either the oviduct or uterus. Later stages are collected by opening the uterus. Late preimplantation stages must be handled carefully because their large size (>2 cm in diameter) results in high surface tension within the yolk sac, especially on the delicate avascular area. When handled too roughly, the embryonic vesicle tends to split and then rapidly collapse; such embryos do not reinflate. If embryos are to be cultured, all steps during collection must be carried out aseptically. In all cases, solutions should be prewarmed to 37°C and embryos should be kept warm by use of a heating stage. Standard fixatives, fixation times, and washing protocols used for mouse embryos are also used for tammar embryos. Methods for embedding tammar conceptuses in paraffin wax depend on the size and stage of the conceptus.


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The Tammar Wallaby, Macropus eugenii: A Model Kangaroo for the Study of Developmental and Reproductive Biology
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This article has been cited by other articles:


Home page
 Cold Spring Harb ProtocHome page
D. Hickford, S. Frankenberg, and M. B. Renfree
The Tammar Wallaby, Macropus eugenii: A Model Kangaroo for the Study of Developmental and Reproductive Biology
Cold Spring Harb Protoc, December 1, 2009; 2009(12): 10.1101/pdb.emo137.
[Abstract] [Full Text]

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 Cold Spring Harb ProtocHome page
D. Hickford, S. Frankenberg, and M. B. Renfree
Culturing Tammar Wallaby (Macropus eugenii) Peri-gastrulation Stage Embryos
Cold Spring Harb Protoc, December 1, 2009; 2009(12): 10.1101/pdb.prot5337.
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 Cold Spring Harb ProtocHome page
D. Hickford, S. Frankenberg, and M. B. Renfree
Whole-Mount Immunohistochemical Staining of Tammar Wallaby (Macropus eugenii) Cleavage Stages and Blastocysts
Cold Spring Harb Protoc, December 1, 2009; 2009(12): 10.1101/pdb.prot5339.
[Abstract] [Full Text]

Home page
 Cold Spring Harb ProtocHome page
D. Hickford, S. Frankenberg, and M. B. Renfree
Whole-Mount Immunohistochemical Staining of Tammar Wallaby (Macropus eugenii) Peri-gastrulation Embryos
Cold Spring Harb Protoc, December 1, 2009; 2009(12): 10.1101/pdb.prot5340.
[Abstract] [Full Text]