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  Light microscopy of echinoderm embryos.

Strickland, L., von Dassow, G., Ellenberg, J., Foe, V., Lenart, P., & Burgess, D. (2004). Light microscopy of echinoderm embryos. In C. A. Ettensohn, G. A. Wray, & G. M. Wessel (Eds.), Development of Sea Urchins, Ascidians, and Other Invertebrate Deuterostomes: Experimental Approaches (pp. 371-409). Elsevier. doi:10.1016/S0091-679X(04)74016-9.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0002-1108-2 Version Permalink: http://hdl.handle.net/21.11116/0000-0002-110A-0
Genre: Book Chapter

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 Creators:
Strickland, L., Author
von Dassow, G., Author
Ellenberg, J., Author
Foe, V., Author
Lenart, P.1, Author              
Burgess, D., Author
Affiliations:
1Research Group of Cytoskeletal Dynamics in Oocytes, MPI for Biophysical Chemistry, Max Planck Society, ou_2640691              

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 Abstract: The chapter discusses commonly used procedures for visualizing both fixed and live echinoderm embryos: (1) formaldehyde fixation of cleavage stage sea urchin embryos, (2) staining and imaging fixed embryos, (3) simultaneous fixation and visualization of the actin and microtubule cytoskeletons, (4) observation of live embryos, (5) 4-D imaging of fluorescent markers in live starfish oocytes, (6) protocol for imaging the dynamics of nuclear lamina during nuclear envelope breakdown, and (7) 4-D imaging of dextran entry during nuclear envelope breakdown (NEBD). Several procedures for fixing early sea urchin embryos with formaldehyde, cold methanol, detergents, and buffers are described. For staining and imaging echinoderm embryo, immunofluorescence is a widely used technique, as there are many commercially available antibodies that will cross-react beautifully with endogenous echinoderm proteins. General protocols for the immunofluorescence of formaldehyde-fixed cells and for staining nucleic acids with various small-molecule dyes are provided in the chapter. The embryos of echinoderms are amenable to live observation, accompanied by time-lapse video microscopy. The different aspects of the cells are emphasized using various imaging techniques, such as Differential Interference Contrast (DIC). Live embryos are easily observed as wet mounts on a standard slide or in a perfusion chamber, but the coverslip prevents direct access to the cells. A variety of chamber slides are described for live observation of echinoderm embryos.

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Language(s): eng - English
 Dates: 2006-05-172004
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1016/S0091-679X(04)74016-9
 Degree: -

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Title: Development of Sea Urchins, Ascidians, and Other Invertebrate Deuterostomes: Experimental Approaches
Source Genre: Book
 Creator(s):
Ettensohn, C. A., Editor
Wray, G. A., Editor
Wessel, G. M., Editor
Affiliations:
-
Publ. Info: Elsevier
Pages: xxiv, 883 Volume / Issue: - Sequence Number: - Start / End Page: 371 - 409 Identifier: ISBN: 978-0-12-480278-0

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Title: Methods in Cell Biology
Source Genre: Series
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Publ. Info: -
Pages: - Volume / Issue: 74 Sequence Number: - Start / End Page: - Identifier: -