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Staging egg development of marine copepods with DAPI and PicoGreen®

ScholarsArchive at Oregon State University

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Title Staging egg development of marine copepods with DAPI and PicoGreen®
Names Zirbel, Marnie Jo (creator)
Miller, Charles B. (creator)
Batchelder, Harold P. (creator)
Date Issued 2007 (iso8601)
Abstract Calanoid copepod eggs have a robust, chitinous outer chorion, which makes field-collected, formaldehydefixed
eggs difficult to penetrate with stains or molecular probes. Egg development studies in copepods have
involved physical, chemical, and enzymatic treatments to remove the chorion. We present an efficient, onestep
method for staining copepod eggs with the fluorescent nucleic acid stains DAPI and PicoGreen®. Nuclei in
treated eggs are clearly visible for examination and counting with compound and confocal microscopy, so that
eggs can be rapidly classified with respect to developmental stage. The method is effective for eggs of Calanus,
Metridia, and Centropages. Both stains were effective after 24-h exposure for eggs that were fixed from several
days to 8 years. Early stages are distinguished by complete counts of nuclei. A blastula phase and gastrulation
are distinctive. Calanus pacificus and Calanus marshallae embryos spend more than half the development period
in the later ‘gray ball’ and limb bud stages. Stage classification by this method is useful for studies of copepod
egg mortality in the field.
Genre Article
Topic Marine Copepods
Identifier Zirbel, M. J., Miller, C. B., & Batchelder, H. P. (2007). Staging egg development of marine copepods with DAPI and PicoGreen®. Limnology and Oceanography: Methods, 5, 106-110.

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