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.
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