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Preparation of Daphnia-parasites for light and electron microscopy and DNA extraction

Please cite as: Ebert, D. Web-guide to Daphnia parasites.

There is nothing like the ultimate method to preserve or fix your samples, but we use routinely the following methods and have good experience with them. We did not find a way to fix field sample for later investigation of host and parasite (e.g. for epidemiological studies). Formol-sugar, which is commonly used to preserve Daphnia and other plankton organisms is only good for the hosts. Microparasites cannot be detected easily in this preservative. The best way to screen mass samples of plankton for parasites is to use fresh material. Keeping it at 4°C will allow you to keep it for a few days. The following methods have all been used successfully for light- and electron-microscopy and DNA extraction.

DNA extraction

Later on, DNA is conveniently extracted
with a commercially availabe kit

Fixing samples for DNA analysis

Fixing for light microscopy

Fixing for electron microscope

Recipes

Bouin fixative

Gluteraldehyde fixative

Cacodylate buffer (0.2M)

Used in Gluteraldehyde fixative and as post fix buffer.