Do sample preparation techniques affect the relative abundance of Florisphaera profunda?

We investigated which method among smearing, settling, filtration, and random settling is the most accurate for determining the relative abundance of Florisphaera profunda. This species is included in paleoproductivity indices because it is one of the most important deep-water dwellers. Therefore, an accurate assessment of its relative abundance is key to monitoring variations in thermocline and nutricline depths. The low birefringence and flat polygonal shape of F. profunda may lead to inaccurate estimates of its relative abundance as it is poorly visible under light microscopy (LM), and different sample preparation may affect the number of specimens on each slide. We studied eight samples from the Ocean Drilling Program (ODP) Site 1209B (Shatsky Rise, NW Pacific) from the last 450kyr. Each sample was prepared five times using each technique for a total of 160 slides. Through a rigorous analytical and statistical approach, we demonstrated the high reproducibility of each method and showed good agreement among techniques at low percentages of F. profunda. When the percentage of this species increases, the random settling technique differs from the others. Filtration and random settling preparation techniques were calibrated by spiking samples with microbeads. The ratio of microbeads observed in slides prepared using these two methods reflects bias due to loss of particles in the filtration technique. In addition, a hydrodynamic model for a fluttering and tumbling plate – such as F. profunda nannoliths – is proposed here along with calculated sinking velocities. These findings confirmed the efficiency of the long decantation time proposed for the random settling technique. The analysis of replicates, the validation with microbeads and the estimation of settling velocity of nannofossils in the tube convinced us that the higher relative abundance recorded by the random technique is not due to an imprecision of this method but, on the contrary, reflects the fact that it is more effective. •The relation between sample preparation techniques and F. profunda abundance is investigated.•Four preparation methods are compared and the results are statistically analyzed by ANOVA.•A hydrodynamic model for fluttering and tumbling plates such as nannoliths is proposed.•All methods yield well reproducible data.•Random settling method seems to be the most accurate method.