Techniques for clearing and mounting Collembola from old ethanol collections
Keywords:
gum-chloral mounting media, high throughput slide mounts, clearing agents, DNA extraction solutionAbstract
Soil mesofauna was collected between 1983 and 2000 from a successional site in Bremen, Germany. The recent analysis of Collembola from this collection posed great challenges in clearing of the specimens stored in 70 % ethanol for 15–32 years for microscopical inspection. For this purpose, five media for permanent slide mounts with different clearing properties were tested: Marc André 2, Huether’s E65, Polyvinyl lactophenol (PVA), Rusek’s and Gisin’s mixture. None of these media cleared the old specimens sufficiently, therefore, pretreatment procedures had to be applied. Two out of eight tested pretreatment methods cleared the aged Collembola successfully: treatment in 10 % NaOH solution at 60°C for 10 minutes, which, however, makes the specimens very fragile. Another pretreatment method originally developed for extracting DNA from Collembola cleared the aged specimens very well and kept them sufficiently stable for further handling. For permanent mounts, Marc André 2 was used, as it has good optical properties, is easy to apply and quickly yields ready-to-handle mounts, especially when dealing with high numbers of preparations for ecological research.
Downloads
References
Bellinger, P. F. (1954): Studies of soil fauna with special reference to the Collembola. – Bulletin of the Connecticut Agricultural Experiment Station New Haven 583: 1–66.
Berch, S. M., B. Baumbrough, J. Battigelli, P. Kroeger, N. Strub & L. de Montigny (2001): Preliminary assessment of selected communities of soil organisms under different conifer
species. – British Columbia, Forest Science Program, Research Report 20: 27 pp.
Berendt, R. (1985): Die sekundäre Sukzession der Collembolenfauna eines Ruderalökosystems und ihre Beeinflussung durch Rekultivierung. – Diploma thesis, University of Bremen: 101 pp.
Bernard, E. C. (2006): Redescription of Cosberella conatoa Wray (Collembola: Hypogastruridae) and description of C. lamaralexanderi n. sp. – Proceedings of the Biological Society of Washington 119(2): 269–278.
Brown, P. A. (1997): A review of techniques used in the preparation, curation and conservation of microscope slides at the Natural History Museum, London. – The Biology Curator 10: 1–33.
Brown, P. A. & E. De Boise (2006): Procedures for the Preparation of Whole Insects as Permanent Microscope Slides and For the Remounting of Deteriorating Aphid Slides. – NatSCA News 8: 15–19.
Dunger, W. & H. J. Fiedler (1997): Methoden der Bodenbiologie. – Jena, Gustav Fischer: 539 pp.
Fjellberg, A. (1980): Identification keys to Norwegian Collembola. – Norsk Entomologisk Forening, Ås: 152 pp.
Gisin, H. (1960): Collembolenfauna Europas. – Museum D’ Histoire Naturelle, Geneva: 312 pp.
Glime, J. M. & D. M. Wagner (2013): Laboratory Techniques: Preservation and Permanent Mounts. Chapt. 2-4. – In: Glime, J. M. (ed.): Bryophyte Ecology. Volume 3. Methods. – Ebook sponsored by Michigan Technological University and the International Association of Bryologists [www.bryoecol.mtu.edu, last updated 5 September 2013].
Greenslade, P. (2010): The aquatic Springtails (Insecta: Collembola) of South Africa. – Australian National University [http://www.ru.ac.za/media/rhodesuniversity/resources/martin/acollembola.html].
Gunter, M. & A. P. Brown (2004): Notes on Conservation Tests of Failing Collembola (Insecta) Micro-Slide Mounts. – NatSCA news 4: 35–36.
Halliday, R. B. (1994): Microscope slide mounting media - results of informal survey. – Acarology List, revised edition 15 June 1994 [http://www.nhm.ac.uk/hosted-sites/acarology/archive/summary.html].
Huether, W. (1993): Sammel- und Präparationsmethoden für Bodenarthropoden. – Abhandlungen und Berichte des Naturkundemuseums Görlitz 66, 6: 1–11.
Koehler, H. (1998): Secondary succession of soil mesofauna: a thirteen year study. – Applied Soil Ecology 9: 81–86.
Koehler, H. (2000): Natural regeneration and succession: results from a 13 years study with reference to mesofauna and vegetation, and implications for management. – Landscape and Urban Planning 51: 123–130.
Koehler, H. & J. Müller (2003): Entwicklung der Biodiversität während einer 20 jährigen Sukzession als Grundlage für Managementmaßnahmen (BMBF report, s.c. 01 LC 0005): 241 pp.
Laird, P. W., A. Zijderveld, K. Linders, M. A. Rudnicki, R. Jaenisch & A. Berns (1991): Simplified Mammalian DNA isolation procedure. – Nucleic Acids Research 19: 4293.
Mari-Mutt, J. (1979): A revision of the genus Dicranocentrus Schött (Insecta: Collembola: Entomobryidae). – Bulletin of the University of Puerto Rico, Agricultural Experiment Station 259: 41–79.
Martin, J. E. H. (1977): Collecting, Preparing and Preserving Insects, Mites, and Spiders. – The Insects and Arachnids of Canada, Vol. 1. – Agriculture Canada: 182 pp.
Rusek, J. & P. Greenslade (1992): Report of round table discussion on methods of mounting Collembola. – Finnish museum of natural history, University of Helsinki: 6 pp.
Rondon, S. & M. Corp (2015): Preserving insects and related arthropods. – Oregon State University [http://extension.oregonstate.edu/umatilla/sites/default/files/PRESERVING__INSECTS.pdf].
Sambrook, J. & D. W. Russell (2001): Molecular Cloning: A laboratory manual, 3rd edition. – Cold Spring Harbor Laboratory Press: 622 pp.
Schauff, M. E. (1986): Collecting and preserving Insects and Mites: techniques and tools. – Agricultural research service, USDA: 69 pp.
Upton, S. M. (1993): Aqueous gum-chloral slide mounting media: an historical review. – Bulletin of Entomological Research 83: 267–274.
Downloads
Published
Issue
Section
License
Soil Organisms is committed to fair open access publishing. All articles are available online without publication fees. Articles published from Vol. 96 No. 3 (2024) onwards are licensed under the Creative Commons Attribution 4.0 International (CC BY 4.0) license. Articles published from Vol. 80 No. 1 through Vol. 96 No. 2 are available under the previous terms, allowing non-commercial, private, and scientific use.
