Impacts of sample storage time on estimates of phytoplankton abundance: how long is too long?

Abstract Phytoplankton play a key role in marine ecosystems, making the accurate quantification of functional groups/species, using standardized microscopy techniques, essential in many research efforts. In this study we aimed to evaluate the effects of (a) storage time on the abundance of microphytoplankton in samples fixed with acid Lugol’s solution; (b) storage time on the abundance of pico- and nanophytoplankton in frozen microscopy slides; (c) storage time on the abundance of pico- and nanophytoplankton in fixed refrigerated samples and (d) storage protocol (refrigerated versus frozen) on the abundance of pico- and nanophytoplankton. Microphytoplankton were analyzed using inverted microscopy, and pico- and nanophytoplankton using epifluorescence microscopy. Results indicate storage time negatively impacted the abundances of all phytoplankton size classes; however, effects were group-specific. For accurate abundance estimates, we suggest that samples fixed with acid Lugol’s should be analyzed within 30 and 180 days, for dinoflagellates/total microphytoplankton and diatoms, respectively. For picoeukaryotes, glutaraldehyde fixed samples should be kept refrigerated for up to 4 days, and slides should be prepared immediately before observation. It is recommended that authors specify the exact lag times between sample collection, fixation and analysis, to allow the comparability of phytoplankton datasets across different studies and/or monitoring programs.