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. 1998 Aug 10;142(3):775–786. doi: 10.1083/jcb.142.3.775

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Complementation assay. Centrosomes isolated from Drosophila embryos are inactivated by incubation with 2 M KI. The KI-treated centrosomes are allowed to bind to a glass coverslip. The coverslip is washed and blocked with a low-salt, BSA-containing buffer, and then incubated with the extract or fraction to be tested. The extract/fraction is washed away and the coverslip is incubated with rhodamine-labeled tubulin. Any resulting asters are fixed sequentially with glutaraldehyde and methanol. The number of asters per 50 microscope fields (100× objective) is determined by counting samples while viewing through a fluorescence microscope. See Materials and Methods for details.