A simple and rapid flow cytometric method of identifying triploidy in larval fish was developed. Prolarvae (4.6–8.0 mm total length) of walleye (Stizostedion vitreum) were mechanically dissociated into single‐cell suspensions and stained with the metachromatic fluorescent dye acridine orange. Dissociation of the prolarvae, application of a two‐step acridine orange staining procedure, and identification of ploidy level by flow cytometry was accomplished in under 10 min/sample. This procedure could be adapted for other fish species and could be used to identify other levels of polyploidy or mosaicism.
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