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Morphological Confocal Microscopy in Arthropods and the Enhancement of Autofluorescence after Proteinase K Extraction

Published online by Cambridge University Press:  03 December 2010

Antonio G. Valdecasas  and
Angela Abad
Antonio G. Valdecasas*
Affiliation:
Museo Nacional de Ciencias Naturales, CSIC, Department of Biodiversity, 2, 28006 – Madrid, Spain
Angela Abad
Affiliation:
Museo Nacional de Ciencias Naturales, CSIC, Department of Biodiversity, 2, 28006 – Madrid, Spain
*
Corresponding author. E-mail: [email protected]
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Abstract

Procedures to study the molecular and morphological characteristics of microscopic organisms are often incompatible with each other. Therein, the realization of alternatives that make the characterization of these features compatible and simultaneously permit the deposition of the original material as a voucher sample into a reference collection is one of the foremost goals of biodiversity studies. In this study, we show that genomic extraction does not necessarily compromise the detailed study of the external morphology of microscopic organisms, and to do so, we used a group of aquatic mites (Acari, Hydrachnidia) as a test group. Hydrachnidia morphology is difficult to study when specimens have been stored in pure ethanol; however, proteinase K extraction leaves them flexible and easy to dissect, while, at the same time, maintaining all of their diagnostic features intact. Furthermore, autofluorescence is significantly enhanced after proteinase extraction. Our study was conducted with aquatic mites that were stored in absolute ethanol in the field and processed for DNA extraction using a Qiagen QIAamp minikit. Before and after molecular extraction, a laser scanning confocal microscopy morphological examination was carried out.

Keywords

LSCMproteinase Kautofluorescence enhancementAcaridiagnostic morphology
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 17 , Issue 1 , February 2011 , pp. 109 - 113
Copyright
Copyright © Microscopy Society of America 2011

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References

REFERENCES

Barr, D. (1973). Methods for the collection, preservation and study of water mites (Acari:Parasitengona). Life Sciences Miscellaneous Publications, Royal Ontario Museum, pp. 1–28. Toronto: University of Toronto.Google Scholar
Chapman, A.D. (2009). Numbers of Living Species in Australia and the World, 2nd ed. Toowoomba, Australia: Australian Biodiversity Information Services.Google Scholar
Cook, D.R. (1974). Water mite genera and subgenera. Memoirs of the American Entomological Institute 21, 1–860.Google Scholar
Dabert, J., Ehrnsberger, R. & Dabert, M. (2008). Glaucalges tytonis sp. n. (Analgoidea, Xolalgidae) from the barn owl Tyto alba (Strigiformes, Tytonidae): Compiling morphology with DNA barcode data for taxon descriptions in mites (Acari). Zootaxa 1719, 41–52.Google Scholar
Gerecke, R. (Ed.) (2007). Chelicerata: Araneae, Acari I. Süsswasserfauna von Mitteleuropa 7/2-1. München, Germany: Spektrum Akademischer Verlag.Google Scholar
Giribet, G. (2010). A new dimension in combining data? The use of morphology and phylogenomic data in metazoan systematics. Acta Zool 91, 11–19.CrossRefGoogle Scholar
Krantz, G.W. & Walter, D.E. (Eds.) (2009). A Manual of Acarology, 3rd ed. Lubbock, TX: Texas Tech University Press.Google Scholar
Lardeux, F., Ung, A. & Chebret, M. (2000). Spectrofluorometers are not adequate for aging Aedes and Culex (Diptera: Culicidae) using pteridine fluorescence. J Med Entomol 37, 769–773.CrossRefGoogle Scholar
Lee, S., Brown, R.L. & Monroe, W. (2009). Use of confocal laser scanning microscopy in systematics of insects with a comparison of fluorescence from different stains. System Entomol 34, 10–14.CrossRefGoogle Scholar
Martin, P., Dabert, M. & Dabert, J. (2010). Molecular evidence for species separation in the water mite Hygrobates nigromaculatus Lebert, 1879 (Acari, Hydrachnidia): Evolutionary consequences of the loss of larval parasitism. Aquatic Sci 72, 347–360.CrossRefGoogle Scholar
Neff, D., Frazier, S.F., Quimby, L., Wang, R. & Zill, S. (2000). Identification of resilin in the leg of cockroach, Periplaneta americana: Confirmation by a simple method using pH dependence of UV fluorescence. Arthropod Struct Develop 29, 75–83.CrossRefGoogle ScholarPubMed
Newell, I.M. (1947). A systematic and ecological study of the Halacaridae of Eastern North America. Bull Bingham Oceanog Collection 10(3), 1–232.Google Scholar
Roy, L., Dowling, A.P.G., Chauve, C.M. & Buronfosse, T. (2009a). Delimiting species boundaries within Dermanyssus Dugès, 1834 (Acari:Dermanyssidae) using a total evidence approach. Molec Phylogen Evol 50, 446–470.CrossRefGoogle ScholarPubMed
Roy, L., Dowling, A.P.G., Chauve, C.M., Lesna, I., Sabelis, M.W. & Buronfosse, T. (2009b). Molecular phylogenetic assessment of host range in five Dermanyssus species. Exp Appl Acarol 48, 115–142.CrossRefGoogle ScholarPubMed
Thorp, J.H. & Covich, A.P. (Eds.) (2001). Ecology and Classification of North American Freshwater Invertebrates, 2nd ed. San Diego, CA: Academic Press.Google Scholar
Valdecasas, A.G. (2008). Confocal microscopy applied to water mite taxonomy with the description of a new genus of Axonopsinae (Acari, Parasitengona, Hydrachnidia) from Central America. Zootaxa 1820, 41–48.CrossRefGoogle Scholar