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Use of freeze-cracking in ontogenetic research in Macrostomum lignano (Macrostomida, Rhabditophora)
Willems, M.; Boone, M.; Couvreur, M.; De Mulder, K.; Van Ranst, J.; Artois, T.; Borgonie, G. (2009). Use of freeze-cracking in ontogenetic research in Macrostomum lignano (Macrostomida, Rhabditophora). Dev. Genes Evol. 219(5): 273-279. hdl.handle.net/10.1007/s00427-009-0284-x
In: Development, Genes and Evolution. Springer: Berlin; Heidelberg. ISSN 0949-944X, more
Peer reviewed article  

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Keywords
    Platyhelminthes [WoRMS]; Marine
Author keywords
    Platyhelminthes; Freeze-cracking; Embryonic development; Eggshell; Immunohistochemistry

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Abstract
    A method for studying whole mount flatworm embryos based on freeze-cracking of the eggs is described. This method allows successful immunohistological and immunocytological studies of whole mount embryos. It does not require the use of sharpened needles or a microinjection system to puncture the eggshell. Moreover, this method is more practical and less time-consuming than classical puncturing and much cheaper than the use of a microinjection system. The advantages of this method are illustrated by results of several immunolocalisation experiments in the macrostomid flatworm Macrostomum lignano. The optimal procedure and crucial steps for this method are discussed.

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