Hostname: page-component-cd9895bd7-fscjk Total loading time: 0 Render date: 2024-12-26T20:25:10.819Z Has data issue: false hasContentIssue false

A Simple and Efficient Method to Observe Internal Structures of Helminths by Scanning Electron Microscopy

Published online by Cambridge University Press:  22 October 2013

Fernando Antônio de O. Adnet
Affiliation:
Laboratório de Biologia de Helmintos Otto Wucherer, Centro de Ciências da Saúde, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Bloco G, Ilha do Fundão, 21949-900 Rio de Janeiro, Brazil Laboratório de Ultraestrutura Celular Hertha Meyer, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Centro de Ciências da Saúde, Bloco G, Ilha do Fundão, 21949-900 Rio de Janeiro, Brazil
Júlia P. Gonçalves
Affiliation:
Laboratório de Biologia de Helmintos Otto Wucherer, Centro de Ciências da Saúde, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Bloco G, Ilha do Fundão, 21949-900 Rio de Janeiro, Brazil Laboratório de Ultraestrutura Celular Hertha Meyer, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Centro de Ciências da Saúde, Bloco G, Ilha do Fundão, 21949-900 Rio de Janeiro, Brazil
Wanderley de Souza
Affiliation:
Laboratório de Ultraestrutura Celular Hertha Meyer, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Centro de Ciências da Saúde, Bloco G, Ilha do Fundão, 21949-900 Rio de Janeiro, Brazil Instituto Nacional de Biologia Estrutural e Bioimagens, Universidade Federal do Rio de Janeiro, Brazil, Bloco G, Ilha do Fundão, 21949-900 Rio de Janeiro, Brazil Instituto Nacional de Metrologia, Qualidade e Tecnologia-Inmetro Bloco G, Ilha do Fundão, 21949-900 Rio de Janeiro, Brazil
Márcia Attias*
Affiliation:
Laboratório de Ultraestrutura Celular Hertha Meyer, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Centro de Ciências da Saúde, Bloco G, Ilha do Fundão, 21949-900 Rio de Janeiro, Brazil
*
*Corresponding author. E-mail: [email protected]; [email protected]
Get access

Abstract

Morphological studies of helminths by scanning electron microscopy are generally limited to the external topography of the organisms. In this work, we present a simple technique using ethanol as a cryoprotectant without postfixation in osmium tetroxide that allows for observation of the inner organization of helminths and preserves cellular structures. We tested the technique in three helminths: Echinostoma paraensei, Cruzia tentaculata, and Hassalstrongylus epsilon. The results show that this technique could potentially be applied to study the morphology, ultrastructure, and taxonomy of helminths.

Type
Biomedical and Biological Applications
Copyright
Copyright © Microscopy Society of America 2013 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Adnet, F.A., Anjos, D.H., Menezes-Oliveira, A. & Lanfredi, R.M. (2009). Further description of Cruzia tentaculata (Rudolphi, 1819) Travassos, 1917 (Nematoda: Cruzidae) by light and scanning electron microscopy. Parasitol Res 104, 12071211.CrossRefGoogle ScholarPubMed
Apkarian, R. & Curtis, J.C. (1981). SEM cryofracture study of ovarian follicles of immature rats. Scan Electron Microsc 72, 165172.Google Scholar
Araújo, A.C., Figueredo-Silva, J., Souto-Padrón, T., Dreyer, G., Norões, J. & De Souza, W. (1995). Scanning electron microscopy of adult Wuchereria bancrofti (Nematoda: Filarioidea). J Parasitol 81, 468474.CrossRefGoogle Scholar
De Moraes Neto, A.H., Lanfredi, R.M. & De Souza, W. (1997). Litomosoides chagasfilhoi sp. nov. (Nematoda: Filarioidea) parasitizing the abdominal cavity of Akodon cursor (Winge, 1887) (Rodentia: Muridae) from Brazil. Parasitol Res 83, 137143.CrossRefGoogle ScholarPubMed
Digiani, M.C., Navone, G.T. & Durette-Desset, M.C. (2007). The systematic position of some nippostrongyline nematodes (Trichostrongylina: Heligmosomoidea) parasitic in Argentinean sigmodontine rodents. Syst Parasitol 67, 8792.CrossRefGoogle ScholarPubMed
Ferraz, J., Souza, J., Costa-Silva, M., Torres, E., Santana, A., Lanfredi, R., Maldonado, A. & Garcia, J. (2012). Effect of praziquantel on adult Echinostoma paraensei worms in experimentally infected mice. Parasitol Res 111, 143148.CrossRefGoogle ScholarPubMed
Fukudome, H. & Tanaka, K. (1986). A method for observing intracellular structure of free cells by scanning electron microscopy. J Microsc 141(Pt 2), 171178.CrossRefGoogle ScholarPubMed
Gonçalves, J.P., Oliveira-Menezes, A., Maldonado Junior, A., Carvalho, T.M. & de Souza, W. (2013). Evaluation of Praziquantel effects on Echinostoma paraensei ultrastructure. Vet Parasitol 194, 1625.CrossRefGoogle ScholarPubMed
Hotchkiss, A.V.J., Martin, V.J. & Apkarian, R.P. (1984). A scanning electron microscopic surface and cryofracture study of development in the Planulae of the hydrozoan, Pennaria tiarella . Scan Electron Microsc 2, 717727.Google Scholar
Humphreys, W.J., Spurlock, B.O. & Johnson, J.S. (1973). Critical point drying of freeze-fractured tissue for scanning electron microscopy. In Proceedings of Thirty-First Annual EMSA Meeting, New Orleans, LA, August 14–17, 1973, pp. 452453. Electron Microscopy Society of America.Google Scholar
Humphreys, W.J., Spurlock, B.O. & Johnson, J.S. (1974). Critical point drying of ethanol infiltrated cryofractured biological specimens for scanning electron microscopy. J Electron Microsc 1, 275282.Google Scholar
Jamjoom, M.B. (2007). Review on electron microscopy in taxonomy and biology of parasitic Nemathelminthes. J Egypt Soc Parasitol 37, 87105.Google ScholarPubMed
Jongebloed, W.L., Stokroos, I., Van der Want, J.J. & Kalicharan, D. (1999). Non-coating fixation techniques or redundancy of conductive coating, low kV FE-SEM operation and combined SEM/TEM of biological tissues. J Microsc 193, 158170.CrossRefGoogle ScholarPubMed
Lee, D.L. (1969). Changes in adult Nippostrongylus brasiliensis during the development of immunity to this nematode in rats. Parasitology 59, 2939.CrossRefGoogle ScholarPubMed
Lichtenfels, J.R., Murrell, K.D. & Pilitt, P.A. (1983). Comparison of three subspecies of Trichinella spiralis by scanning electron microscopy. J Parasitol 69, 11311140.CrossRefGoogle ScholarPubMed
Lorsuwannarat, N., Saowakon, N., Ramasoota, P., Wanichanon, C. & Sobhon, P. (2013). The anthelmintic effect of Plumbagin on Schistosoma mansoni . Exp Parasitol 133, 1827.CrossRefGoogle ScholarPubMed
Oliveira-Menezes, A., De Souza, W. & Lanfredi, R.M. (2003). Cuticular architecture of Hassalstrongylus epsilon (Nematoda: Trichostrongyloidea). Parasitol Res 90, 280286.CrossRefGoogle ScholarPubMed
Rinaldi, V., Ravazzoni, L., Magno, M.S. & Djaczenko, W. (1973). Scanning electron microscopy of the tegument of Taenia saginata . Rev Biol 66, 417424.Google ScholarPubMed
Riva, A., Faa, G., Loffredo, F., Piludu, M. & Riva, F.T. (1999). An improved OsO4 maceration method for the visualization of internal structures and surfaces in human bioptic specimens by high resolution scanning electron microscopy. Scanning Microscopy 13, 111122.Google Scholar
Roberts, L. & Janovy, S. (2008). Foundations of Parasitology, 8th ed. New York: McGraw-Hill Companies.Google Scholar
Souza, J.G., Garcia, J.S., Manso, P.P., Neves, R.H., Maldonado, A. Jr. & Machado-Silva, J.R. (2011). Development of the reproductive system of Echinostoma paraensei in Mesocricetus auratus analyzed by light and confocal scanning laser microscopy. Exp Parasitol 128, 341346.CrossRefGoogle ScholarPubMed
Taylor, J. (2008). Cryofracture or freeze-fracture, a method to expose internal tissue surfaces and cell surfaces for viewing in the scanning electron microscope. Micros Today 16, 5658.CrossRefGoogle Scholar
Torres, E.J., Nascimento, A.P., Menezes, A.O., Garcia, J., Dos Santos, M.A., Maldonado, A. Jr., Miranda, K., Lanfredi, R.M. & De Souza, W. (2011). A new species of Trichuris from Thrichomys apereoides (Rodentia: Echimyidae) in Brazil: Morphological and histological studies. Vet Parasitol 176, 226235.CrossRefGoogle ScholarPubMed
Watson, B.D. (1965). The fine structure of the body-wall and the growth of the cuticle in the adult nematode Ascaris lumbricoides. Quart J Micr Sci 106, 7581.Google Scholar