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Comparative Effects of Event Detection Methods on the Analysis and Interpretation of Ca²⁺ Imaging Data.

ABSTRACT: Calcium imaging has gained substantial popularity as a tool to profile the activity of multiple simultaneously active cells at high spatiotemporal resolution. Among the diverse approaches to processing of Ca²⁺ imaging data is an often subjective decision of how to quantify baseline fluorescence or F ₀. We examine the effect of popular F ₀ determination methods on the interpretation of neuronal and astrocyte activity in a single dataset of rats trained to self-administer intravenous infusions of cocaine and compare them with an F ₀-independent wavelet ridgewalking event detection approach. We find that the choice of the processing method has a profound impact on the interpretation of widefield imaging results. All of the dF/F ₀ thresholding methods tended to introduce spurious events and fragment individual transients, leading to smaller calculated event durations and larger event frequencies. Analysis of simulated datasets confirmed these observations and indicated substantial intermethod variability as to the events classified as significant. Additionally, most dF/F ₀ methods on their own were unable to adequately account for bleaching of fluorescence, although the F ₀ smooth approach and the wavelet ridgewalking algorithm both did so. In general, the choice of the processing method led to dramatically different quantitative and sometimes opposing qualitative interpretations of the effects of cocaine self-administration both at the level of individual cells and at the level of cell networks. Significantly different distributions of event duration, amplitude, frequency, and network measures were found across the majority of dF/F ₀ approaches. The wavelet ridgewalking algorithm broadly outperformed dF/F ₀-based methods for both neuron and astrocyte recordings. These results indicate the need for heightened awareness of the limitations and tendencies associated with decisions to use particular Ca²⁺ image processing pipelines. Both quantification and interpretation of the effects of experimental manipulations are strongly sensitive to such decisions.

SUBMITTER: Neugornet A

PROVIDER: S-EPMC8032960 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundAs a hot method in machine learning field, the forests approach is an attractive alternative approach to Cox model. Random survival forests (RSF) methodology is the most popular survival forests method, whereas its drawbacks exist such as a selection bias towards covariates with many possible split points. Conditional inference forests (CIF) methodology is known to reduce the selection bias via a two-step split procedure implementing hypothesis tests as it separates the variable selection and splitting, but its computation costs too much time. Random forests with maximally selected rank statistics (MSR-RF) methodology proposed recently seems to be a great improvement on RSF and CIF.MethodsIn this paper we used simulation study and real data application to compare prediction performances and variable selection performances among three survival forests methods, including RSF, CIF and MSR-RF. To evaluate the performance of variable selection, we combined all simulations to calculate the frequency of ranking top of the variable importance measures of the correct variables, where higher frequency means better selection ability. We used Integrated Brier Score (IBS) and c-index to measure the prediction accuracy of all three methods. The smaller IBS value, the greater the prediction.ResultsSimulations show that three forests methods differ slightly in prediction performance. MSR-RF and RSF might perform better than CIF when there are only continuous or binary variables in the datasets. For variable selection performance, When there are multiple categorical variables in the datasets, the selection frequency of RSF seems to be lowest in most cases. MSR-RF and CIF have higher selection rates, and CIF perform well especially with the interaction term. The fact that correlation degree of the variables has little effect on the selection frequency indicates that three forest methods can handle data with correlation. When there are only continuous variables in the datasets, MSR-RF perform better. When there are only binary variables in the datasets, RSF and MSR-RF have more advantages than CIF. When the variable dimension increases, MSR-RF and RSF seem to be more robustthan CIF CONCLUSIONS: All three methods show advantages in prediction performances and variable selection performances under different situations. The recent proposed methodology MSR-RF possess practical value and is well worth popularizing. It is important to identify the appropriate method in real use according to the research aim and the nature of covariates.

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Comparative Effects of Event Detection Methods on the Analysis and Interpretation of Ca²⁺ Imaging Data.

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Comparative Effects of Event Detection Methods on the Analysis and Interpretation of Ca2+ Imaging Data.

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Comparative Effects of Event Detection Methods on the Analysis and Interpretation of Ca²⁺ Imaging Data.