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Semiquantitative Confocal Laser Scanning Microscopy Applied to Marine Invertebrate Ecotoxicology

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Abstract

Confocal laser scanning microscopy (CLSM) represents a powerful, but largely unexplored ecotoxicologic tool for rapidly assessing in vivo effects of toxicants on marine invertebrate embryo quality and development. We describe here a new semiquantitative CLSM approach for assessing relative yolk quantity in marine invertebrate embryos (harpacticoid copepods) produced by parents reared from hatching to adult in the polycylic aromatic hydrocarbon chrysene. This method is based on fluorogenic labeling of embryo yolk and subsequent statistical analysis of areal pixel intensities over multiple Z-series using a general linear model (GLM)–nested analysis of variance. The fluorescent yolk-labeling method described here was able to detect statistically significant differences in yolk concentrations in marine copepod (Amphiascus tenuiremis) eggs or embryos from females exposed to ultraviolet light and chrysene-contaminated sediments. Yolk intensities in embryos from females cultured throughout their life cycles in clean sediments were statistically identical with or without UV exposure. In constrast, yolk intensities in embryos of females cultured throughout their life cycle in chrysene-contaminated sediments were significantly higher in the non-UV-exposed treatment with chrysene at 2500 ng/g sediment (65.7% higher) and the UV-exposed treatment with chrysene at 500 ng/g sediment (76.6% higher).

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Acknowledgments

This research was funded by the U.S. Environmental Protection Agency through grant R-82-7397 to T. Chandler, G. Scott, J. Ferry, E. Wirth, J. Quattro, and M. Fulton. This research has not been subjected to the Agency’s required peer and policy review and therefore does not necessarily reflect the views of the Agency, and no endorsement should be inferred.

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  1. Department of Environmental Health Sciences, Norman J. Arnold School of Public Health, 800 Sumter Street, Room 311, University of South Carolina, Columbia, SC 29208, USA

    G. Thomas Chandler & David C. Volz

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  1. G. Thomas Chandler
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  2. David C. Volz
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Correspondence to G. Thomas Chandler.

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Chandler, G., Volz, D.C. Semiquantitative Confocal Laser Scanning Microscopy Applied to Marine Invertebrate Ecotoxicology . Mar. Biotechnol. 6, 128–137 (2004). https://doi.org/10.1007/s10126-002-0105-y

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