Many experiments such as transfection of cells and organisms, live imaging and certain biochemical assays, require isolation of live, viable bacteria from insects.
This protocol was developed to isolate Wolbachia from adult Drosophila, but it can be adapted for other insects. In some insects leg removal prior to isolation facilitates hemolymph extrusion 1. This is not necessary for Drosophila.
Conventionally, for transfection experiments, Wolbachia are isolated from embryonic cytoplasm 2-5. In comparison, the main advantages of this method are:
Better elimination of debris from ultra-filtration.
Faster; requires less manipulation.
Less yolk is co-purified with the bacteria. Due to the associated auto fluorescence, yolk becomes an issue for imaging (see also “Live imaging of Wolbachia”).