Spatial smoothing is an essential pre-processing step in the process of analysing functional magnetic resonance imaging (fMRI) data, both during an experimental task or during resting-state fMRI (rsfMRI). The main benefit of this spatial smoothing step is to artificially increase the signal-to-noise ratio of the fMRI signal. Previous fMRI studies have investigated the impact of spatial smoothing on task fMRI data, while rsfMRI studies usually apply the same analytical process used for the task data. However, this study investigates changes in different rsfMRI analyses, such as ROI-to-ROI, seed-to-voxels and ICA analyses. Nineteen healthy volunteers were scanned using rsfMRI with three applied smoothing kernels: 0 mm, 4 mm and 8 mm. Appropriate statistical comparisons were made. The findings showed that spatial smoothing has a greater effect on rsfMRI data when analysed using seed-to-voxel-based analysis. The effect was less pronounced when analysing data using ROI-ROI or ICA analyses. The results demonstrated that even when analysing the data without the application of spatial smoothing, the results were significant compared with data analysed using a typical smoothing kernel. However, data analysed with lower-smoothing kernels produced greater negative correlations, particularly with the ICA analysis. The results suggest that a medium smoothing kernel (around 4 mm) may be preferable, as it is comparable with the 8 mm kernel in all of the analyses performed. It is also recommended that the researchers consider analysing the data using two different smoothing kernels, as this will help to confirm the significance of the results and avoid overestimating the findings.
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