Long-Term and Meditation-Specific Modulations of Brain Connectivity Revealed Through Multivariate Pattern Analysis.

Neuroimaging studies have provided evidence that extensive meditation practice modifies the functional and structural properties of the human brain, such as large-scale brain region interplay. However, it remains unclear how different meditation styles are involved in the modulation of these large-scale brain networks. Here, using machine learning and fMRI functional connectivity, we investigated how focused attention and open monitoring meditation styles impact large-scale brain networks. Specifically, we trained a classifier to predict the meditation style in two groups of subjects: expert Theravada Buddhist monks and novice meditators. We showed that the classifier was able to discriminate the meditation style only in the expert group. Additionally, by inspecting the trained classifier, we observed that the Anterior Salience and the Default Mode networks were relevant for the classification, in line with their theorized involvement in emotion and self-related regulation in meditation. Interestingly, results also highlighted the role of specific couplings between areas crucial for regulating attention and self-awareness as well as areas related to processing and integrating somatosensory information. Finally, we observed a larger involvement of left inter-hemispheric connections in the classification. In conclusion, our work supports the evidence that extensive meditation practice modulates large-scale brain networks, and that the different meditation styles differentially affect connections that subserve style-specific functions.

Spectral signature of attentional reorienting in the human brain.

As we move in the environment, attention shifts to novel objects of interest based on either their sensory salience or behavioral value (reorienting). This study measures with magnetoencephalography (MEG) different properties (amplitude, onset-to-peak duration) of event-related desynchronization/synchronization (ERD/ERS) of oscillatory activity during a visuospatial attention task designed to separate activity related to reorienting vs. maintaining attention to the same location, controlling for target detection and response processes. The oscillatory activity was measured both in fMRI-defined regions of interest (ROIs) of the dorsal attention (DAN) and visual (VIS) networks, previously defined as task-relevant in the same subjects, or whole-brain in a pre-defined set of cortical ROIs encompassing the main brain networks. Reorienting attention (shift cues) as compared to maintaining attention (stay cues) produced a temporal sequence of ERD/ERS modulations at multiple frequencies in specific anatomical regions/networks. An early (∼330 ms), stronger, transient theta ERS occurred in task-relevant (DAN, VIS) and control networks (VAN, CON, FPN), possibly reflecting an alert/reset signal in response to the cue. A more sustained, behaviorally relevant, low-beta band ERD peaking ∼450 ms following shift cues (∼410 for stay cues) localized in frontal and parietal regions of the DAN. This modulation is consistent with a control signal re-routing information across visual hemifields. Contralateral vs. ipsilateral shift cues produced in occipital visual regions a stronger, sustained alpha ERD (peak ∼470 ms) and a longer, transient high beta/gamma ERS (peak ∼490 ms) related to preparatory visual modulations in advance of target occurrence. This is the first description of a cascade of oscillatory processes during attentional reorienting in specific anatomical regions and networks. Among these processes, a behaviorally relevant beta desynchronization in the FEF is likely associated with the control of attention shifts.

The impact of improved MEG-MRI co-registration on MEG connectivity analysis.

Co-registration between structural head images and functional MEG data is needed for anatomically-informed MEG data analysis. Despite the efforts to minimize the co-registration error, conventional landmark- and surface-based strategies for co-registering head and MEG device coordinates achieve an accuracy of typically 5-10 mm. Recent advances in instrumentation and technical solutions, such as the development of hybrid ultra-low-field (ULF) MRI-MEG devices or the use of 3D-printed individualized foam head-casts, promise unprecedented co-registration accuracy, i.e., 2 mm or better. In the present study, we assess through simulations the impact of such an improved co-registration on MEG connectivity analysis. We generated synthetic MEG recordings for pairs of connected cortical sources with variable locations. We then assessed the capability to reconstruct source-level connectivity from these recordings for 0-15-mm co-registration error, three levels of head modeling detail (one-, three- and four-compartment models), two source estimation techniques (linearly constrained minimum-variance beamforming and minimum-norm estimation MNE) and five separate connectivity metrics (imaginary coherency, phase-locking value, amplitude-envelope correlation, phase-slope index and frequency-domain Granger causality). We found that beamforming can better take advantage of an accurate co-registration than MNE. Specifically, when the co-registration error was smaller than 3 mm, the relative error in connectivity estimates was down to one-third of that observed with typical co-registration errors. MNE provided stable results for a wide range of co-registration errors, while the performance of beamforming rapidly degraded as the co-registration error increased. Furthermore, we found that even moderate co-registration errors (>6 mm, on average) essentially decrease the difference of four- and three- or one-compartment models. Hence, a precise co-registration is important if one wants to take full advantage of highly accurate head models for connectivity analysis. We conclude that an improved co-registration will be beneficial for reliable connectivity analysis and effective use of highly accurate head models in future MEG connectivity studies.

Alpha and alpha-beta phase synchronization mediate the recruitment of the visuospatial attention network through the Superior Longitudinal Fasciculus.

It is well known that attentional selection of relevant information relies on local synchronization of alpha band neuronal oscillations in visual cortices for inhibition of distracting inputs. Additionally, evidence for long-range coupling of neuronal oscillations between visual cortices and regions engaged in the anticipation of upcoming stimuli has been more recently provided. Nevertheless, on the one hand the relation between long-range functional coupling and anatomical connections is still to be assessed, and, on the other hand, the specific role of the alpha and beta frequency bands in the different processes underlying visuo-spatial attention still needs further clarification. We address these questions using measures of linear (frequency-specific) and nonlinear (cross-frequency) phase-synchronization in a cohort of 28 healthy subjects using magnetoencephalography. We show that alpha band phase-synchronization is modulated by the orienting of attention according to a parieto-occipital top-down mechanism reflecting behavior, and its hemispheric asymmetry is predicted by volume's asymmetry of specific tracts of the Superior-Longitudinal-Fasciculus. We also show that a network comprising parietal regions and the right putative Frontal-Eye-Field, but not the left, is recruited in the deployment of spatial attention through an alpha-beta cross-frequency coupling. Overall, we demonstrate that the visuospatial attention network features subsystems indexed by characteristic spectral fingerprints, playing different functional roles in the anticipation of upcoming stimuli and with diverse relation to fiber tracts.

A Comparative Study of the Robustness of Frequency-Domain Connectivity Measures to Finite Data Length.

In this work we use numerical simulation to investigate how the temporal length of the data affects the reliability of the estimates of brain connectivity from EEG time-series. We assume that the neural sources follow a stable MultiVariate AutoRegressive model, and consider three connectivity metrics: imaginary part of coherency (IC), generalized partial directed coherence (gPDC) and frequency-domain granger causality (fGC). In order to assess the statistical significance of the estimated values, we use the surrogate data test by generating phase-randomized and autoregressive surrogate data. We first consider the ideal case where we know the source time courses exactly. Here we show how, expectedly, even exact knowledge of the source time courses is not sufficient to provide reliable estimates of the connectivity when the number of samples gets small; however, while gPDC and fGC tend to provide a larger number of false positives, the IC becomes less sensitive to the presence of connectivity. Then we proceed with more realistic simulations, where the source time courses are estimated using eLORETA, and the EEG signal is affected by biological noise of increasing intensity. Using the ideal case as a reference, we show that the impact of biological noise on IC estimates is qualitatively different from the impact on gPDC and fGC.

Mapping Brain Activity with Electrocorticography: Resolution Properties and Robustness of Inverse Solutions.

Electrocorticography (ECoG) is an electrophysiological technique that records brain activity directly from the cortical surface with high temporal (ms) and spatial (mm) resolution. Its major limitations are in the high invasiveness and in the restricted field-of-view of the electrode grid, which partially covers the cortex. To infer brain activity at locations different from just below the electrodes, it is necessary to solve the electromagnetic inverse problem. Limitations in the performance of source reconstruction algorithms from ECoG have been, to date, only partially addressed in the literature, and a systematic evaluation is still lacking. The main goal of this study is to provide a quantitative evaluation of resolution properties of widely used inverse methods (eLORETA and MNE) for various ECoG grid sizes, in terms of localization error, spatial dispersion, and overall amplitude. Additionally, this study aims at evaluating how the use of simultaneous electroencephalography (EEG) affects the above properties. For these purposes, we take advantage of a unique dataset in which a monkey underwent a simultaneous recording with a 128 channel ECoG grid and an 18 channel EEG grid. Our results show that, in general conditions, the reconstruction of cortical activity located more than 1 cm away from the ECoG grid is not accurate, since the localization error increases linearly with the distance from the electrodes. This problem can be partially overcome by recording simultaneously ECoG and EEG. However, this analysis enlightens the necessity to design inverse algorithms specifically targeted at taking into account the limited field-of-view of the ECoG grid.

Disclosing large-scale directed functional connections in MEG with the multivariate phase slope index.

The phase slope index (PSI) is a method to disclose the direction of frequency-specific neural interactions from magnetoencephalographic (MEG) time series. A fundamental property of PSI is that of vanishing for linear mixing of independent neural sources. This property allows PSI to cope with the artificial instantaneous connectivity among MEG sensors or brain sources induced by the field spread. Nevertheless, PSI is limited by being a bivariate estimator of directionality as opposite to the multidimensional nature of brain activity as revealed by MEG. The purpose of this work is to provide a multivariate generalization of PSI. We termed this measure as the multivariate phase slope index (MPSI). In order to test the ability of MPSI in estimating the directionality, and to compare the MPSI results to those obtained by bivariate PSI approaches based on maximizing imaginary part of coherency and on canonical correlation analysis, we used extensive simulations. We proved that MPSI achieves the highest performance and that in a large number of simulated cases, the bivariate methods, as opposed to MPSI, do not detect a statistically significant directionality. Finally, we applied MPSI to assess seed-based directed functional connectivity in the alpha band from resting state MEG data of 61 subjects from the Human Connectome Project. The obtained results highlight a directed functional coupling in the alpha band between the primary visual cortex and several key regions of well-known resting state networks, e.g. dorsal attention network and fronto-parietal network.

Functional connections between activated and deactivated brain regions mediate emotional interference during externally directed cognition.

Recent evidence shows that task-deactivations are functionally relevant for cognitive performance. Indeed, higher cognitive engagement has been associated with higher suppression of activity in task-deactivated brain regions - usually ascribed to the Default Mode Network (DMN). Moreover, a negative correlation between these regions and areas actively engaged by the task is associated with better performance. DMN regions show positive modulation during autobiographical, social, and emotional tasks. However, it is not clear how processing of emotional stimuli affects the interplay between the DMN and executive brain regions. We studied this interplay in an fMRI experiment using emotional negative stimuli as distractors. Activity modulations induced by the emotional interference of negative stimuli were found in frontal, parietal, and visual areas, and were associated with modulations of functional connectivity between these task-activated areas and DMN regions. A worse performance was predicted both by lower activity in the superior parietal cortex and higher connectivity between visual areas and frontal DMN regions. Connectivity between right inferior frontal gyrus and several DMN regions in the left hemisphere was related to the behavioral performance. This relation was weaker in the negative than in the neutral condition, likely suggesting less functional inhibitions of DMN regions during emotional processing. These results show that both executive and DMN regions are crucial for the emotional interference process and suggest that DMN connections are related to the interplay between externally-directed and internally-focused processes. Among DMN regions, superior frontal gyrus may be a key node in regulating the interference triggered by emotional stimuli.

Concurrent brain responses to separate auditory and visual targets.

In the attentional blink, a target event (T1) strongly interferes with perception of a second target (T2) presented within a few hundred milliseconds. Concurrently, the brain's electromagnetic response to the second target is suppressed, especially a late negative-positive EEG complex including the traditional P3 wave. An influential theory proposes that conscious perception requires access to a distributed, frontoparietal global workspace, explaining the attentional blink by strong mutual inhibition between concurrent workspace representations. Often, however, the attentional blink is reduced or eliminated for targets in different sensory modalities, suggesting a limit to such global inhibition. Using functional magnetic resonance imaging, we confirm that visual and auditory targets produce similar, distributed patterns of frontoparietal activity. In an attentional blink EEG/MEG design, however, an auditory T1 and visual T2 are identified without mutual interference, with largely preserved electromagnetic responses to T2. The results suggest parallel brain responses to target events in different sensory modalities.

Being an agent or an observer: different spectral dynamics revealed by MEG.

Several neuroimaging studies reported that a common set of regions is recruited during action observation and execution and it has been proposed that the modulation of the µ rhythm, in terms of oscillations in the alpha and beta bands might represent the electrophysiological correlate of the underlying brain mechanisms. However, the specific functional role of these bands within the µ rhythm is still unclear. Here, we used magnetoencephalography (MEG) to analyze the spectral and temporal properties of the alpha and beta bands in healthy subjects during an action observation and execution task. We associated the modulation of the alpha and beta power to a broad action observation network comprising several parieto-frontal areas previously detected in fMRI studies. Of note, we observed a dissociation between alpha and beta bands with a slow-down of beta oscillations compared to alpha during action observation. We hypothesize that this segregation is linked to a different sequence of information processing and we interpret these modulations in terms of internal models (forward and inverse). In fact, these processes showed opposite temporal sequences of occurrence: anterior-posterior during action (both in alpha and beta bands) and roughly posterior-anterior during observation (in the alpha band). The observed differentiation between alpha and beta suggests that these two bands might pursue different functions in the action observation and execution processes.

Local and remote effects of transcranial direct current stimulation on the electrical activity of the motor cortical network.

We systematically investigated the effects of cathodal and anodal Transcranial Direct Current Stimulation (CtDCS, AtDCS) on the electric activity of primary motor cortex during a motor task. High-density electroencephalography was used to define the spatial diffusion of tDCS after effects. Ten healthy subjects performed a finger tapping task with the right hand before and after three separate sessions of 20 minutes of Sham, AtDCS or CtDCS over left primary motor cortex (M1). During movement, we found an increment of low alpha band Event-Related Desynchronization (ERD) in bilateral central, frontal areas and in the left inferior parietal region, as well as an increment of beta ERD in fronto-central and parieto-occipital regions, after AtDCs compared to Sham and CtDCS. In the rest pre-movement period, after Sham as well as AtDCS, we documented an increment of low alpha band power over the course of pre- and post-stimulation recording sessions, localized in the sensorimotor and parieto-occipital regions. On the contrary, after CtDCS no increment of low alpha power was found. Finally beta band coherence among signals from left sensorimotor cortex and activity of bilateral parietal, occipital and right frontal regions was higher after AtDCS compared with Sham condition. Similarly, theta coherence with parietal and frontal regions was enhanced after AtDCS. We hypothesize that the local modulation of membrane polarization, as well as long-lasting synaptic modification induced by tDCS over M1, could result in changes of both local band power and functional architecture of the motor network.

A bicoherence approach to analyze multi-dimensional cross-frequency coupling in EEG/MEG data.

We introduce a blockwise generalisation of the Antisymmetric Cross-Bicoherence (ACB), a statistical method based on bispectral analysis. The Multi-dimensional ACB (MACB) is an approach that aims at detecting quadratic lagged phase-interactions between vector time series in the frequency domain. Such a coupling can be empirically observed in functional neuroimaging data, e.g., in electro/magnetoencephalographic signals. MACB is invariant under orthogonal trasformations of the data, which makes it independent, e.g., on the choice of the physical coordinate system in the neuro-electromagnetic inverse procedure. In extensive synthetic experiments, we prove that MACB performance is significantly better than that obtained by ACB. Specifically, the shorter the data length, or the higher the dimension of the single data space, the larger the difference between the two methods.

Mindfulness meditation styles differently modulate source-level MEG microstate dynamics and complexity.

Background: The investigation of mindfulness meditation practice, classically divided into focused attention meditation (FAM), and open monitoring meditation (OMM) styles, has seen a long tradition of theoretical, affective, neurophysiological and clinical studies. In particular, the high temporal resolution of magnetoencephalography (MEG) or electroencephalography (EEG) has been exploited to fill the gap between the personal experience of meditation practice and its neural correlates. Mounting evidence, in fact, shows that human brain activity is highly dynamic, transiting between different brain states (microstates). In this study, we aimed at exploring MEG microstates at source-level during FAM, OMM and in the resting state, as well as the complexity and criticality of dynamic transitions between microstates. Methods: Ten right-handed Theravada Buddhist monks with a meditative expertise of minimum 2,265 h participated in the experiment. MEG data were acquired during a randomized block design task (6 min FAM, 6 min OMM, with each meditative block preceded and followed by 3 min resting state). Source reconstruction was performed using eLORETA on individual cortical space, and then parcellated according to the Human Connect Project atlas. Microstate analysis was then applied to parcel level signals in order to derive microstate topographies and indices. In addition, from microstate sequences, the Hurst exponent and the Lempel-Ziv complexity (LZC) were computed. Results: Our results show that the coverage and occurrence of specific microstates are modulated either by being in a meditative state or by performing a specific meditation style. Hurst exponent values in both meditation conditions are reduced with respect to the value observed during rest, LZC shows significant differences between OMM, FAM, and REST, with a progressive increase from REST to FAM to OMM. Discussion: Importantly, we report changes in brain criticality indices during meditation and between meditation styles, in line with a state-like effect of meditation on cognitive performance. In line with previous reports, we suggest that the change in cognitive state experienced in meditation is paralleled by a shift with respect to critical points in brain dynamics.

Exploring Motor Network Connectivity in State-Dependent Transcranial Magnetic Stimulation: A Proof-of-Concept Study.

State-dependent non-invasive brain stimulation (NIBS) informed by electroencephalography (EEG) has contributed to the understanding of NIBS inter-subject and inter-session variability. While these approaches focus on local EEG characteristics, it is acknowledged that the brain exhibits an intrinsic long-range dynamic organization in networks. This proof-of-concept study explores whether EEG connectivity of the primary motor cortex (M1) in the pre-stimulation period aligns with the Motor Network (MN) and how the MN state affects responses to the transcranial magnetic stimulation (TMS) of M1. One thousand suprathreshold TMS pulses were delivered to the left M1 in eight subjects at rest, with simultaneous EEG. Motor-evoked potentials (MEPs) were measured from the right hand. The source space functional connectivity of the left M1 to the whole brain was assessed using the imaginary part of the phase locking value at the frequency of the sensorimotor µ-rhythm in a 1 s window before the pulse. Group-level connectivity revealed functional links between the left M1, left supplementary motor area, and right M1. Also, pulses delivered at high MN connectivity states result in a greater MEP amplitude compared to low connectivity states. At the single-subject level, this relation is more highly expressed in subjects that feature an overall high cortico-spinal excitability. In conclusion, this study paves the way for MN connectivity-based NIBS.

Temporal dynamics of spontaneous MEG activity in brain networks.

Functional MRI (fMRI) studies have shown that low-frequency (<0.1 Hz) spontaneous fluctuations of the blood oxygenation level dependent (BOLD) signal during restful wakefulness are coherent within distributed large-scale cortical and subcortical networks (resting state networks, RSNs). The neuronal mechanisms underlying RSNs remain poorly understood. Here, we describe magnetoencephalographic correspondents of two well-characterized RSNs: the dorsal attention and the default mode networks. Seed-based correlation mapping was performed using time-dependent MEG power reconstructed at each voxel within the brain. The topography of RSNs computed on the basis of extended (5 min) epochs was similar to that observed with fMRI but confined to the same hemisphere as the seed region. Analyses taking into account the nonstationarity of MEG activity showed transient formation of more complete RSNs, including nodes in the contralateral hemisphere. Spectral analysis indicated that RSNs manifest in MEG as synchronous modulation of band-limited power primarily within the theta, alpha, and beta bands-that is, in frequencies slower than those associated with the local electrophysiological correlates of event-related BOLD responses.

Effective Intracerebral Connectivity in Acute Stroke: A TMS-EEG Study.

Stroke is a major cause of disability because of its motor and cognitive sequelae even when the acute phase of stabilization of vital parameters is overcome. The most important improvements occur in the first 8-12 weeks after stroke, indicating that it is crucial to improve our understanding of the dynamics of phenomena occurring in this time window to prospectively target rehabilitation procedures from the earliest stages after the event. Here, we studied the intracortical excitability properties of delivering transcranial magnetic stimulation (TMS) to the primary motor cortex (M1) of left and right hemispheres in 17 stroke patients who suffered a mono-lateral left hemispheric stroke, excluding pure cortical damage. All patients were studied within 10 days of symptom onset. TMS-evoked potentials (TEPs) were collected via a TMS-compatible electroencephalogram system (TMS-EEG) concurrently with motor-evoked responses (MEPs) induced in the contralateral first dorsal interosseous muscle. Comparison with age-matched healthy volunteers was made by collecting the same bilateral-stimulation data in nine healthy volunteers as controls. Excitability in the acute phase revealed relevant changes in the relationship between left lesioned and contralesionally right hemispheric homologous areas both for TEPs and MEPs. While the paretic hand displayed reduced MEPs compared to the non-paretic hand and to healthy volunteers, TEPs revealed an overexcitable lesioned hemisphere with respect to both healthy volunteers and the contra-lesion side. Our quantitative results advance the understanding of the impairment of intracortical inhibitory networks. The neuronal dysfunction most probably changes the excitatory/inhibitory on-center off-surround organization that supports already acquired learning and reorganization phenomena that support recovery from stroke sequelae.

TMS-Induced Modulation of EEG Functional Connectivity Is Affected by the E-Field Orientation.

Coregistration of transcranial magnetic stimulation (TMS) and electroencephalography (EEG) allows non-invasive probing of brain circuits: TMS induces brain activation due to the generation of a properly oriented focused electric field (E-field) using a coil placed on a selected position over the scalp, while EEG captures the effects of the stimulation on brain electrical activity. Moreover, the combination of these techniques allows the investigation of several brain properties, including brain functional connectivity. The choice of E-field parameters, such as intensity, orientation, and position, is crucial for eliciting cortex-specific effects. Here, we evaluated whether and how the spatial pattern, i.e., topography and strength of functional connectivity, is modulated by the stimulus orientation. We systematically altered the E-field orientation when stimulating the left pre-supplementary motor area and showed an increase of functional connectivity in areas associated with the primary motor cortex and an E-field orientation-specific modulation of functional connectivity intensity.

Looking through the windows: a study about the dependency of phase-coupling estimates on the data length.

Objective. Being able to characterize functional connectivity (FC) state dynamics in a real-time setting, such as in brain-computer interface, neurofeedback or closed-loop neurostimulation frameworks, requires the rapid detection of the statistical dependencies that quantify FC in short windows of data. The aim of this study is to characterize, through extensive realistic simulations, the reliability of FC estimation as a function of the data length. In particular, we focused on FC as measured by phase-coupling (PC) of neuronal oscillations, one of the most functionally relevant neural coupling modes.Approach. We generated synthetic data corresponding to different scenarios by varying the data length, the signal-to-noise ratio (SNR), the phase difference value, the spectral analysis approach (Hilbert or Fourier) and the fractional bandwidth. We compared seven PC metrics, i.e. imaginary part of phase locking value (iPLV), PLV of orthogonalized signals, phase lag index (PLI), debiased weighted PLI, imaginary part of coherency, coherence of orthogonalized signals and lagged coherence.Main results. Our findings show that, for a SNR of at least 10 dB, a data window that contains 5-8 cycles of the oscillation of interest (e.g. a 500-800 ms window at 10 Hz) is generally required to achieve reliable PC estimates. In general, Hilbert-based approaches were associated with higher performance than Fourier-based approaches. Furthermore, the results suggest that, when the analysis is performed in a narrow frequency range, a larger window is required.Significance. The achieved results pave the way to the introduction of best-practice guidelines to be followed when a real-time frequency-specific PC assessment is at target.

Magnetoencephalographic spectral fingerprints differentiate evidence accumulation from saccadic motor preparation in perceptual decision-making.

The understanding of the neurobiological basis of perceptual decision-making has been profoundly shaped by studies in the monkey brain in tandem with mathematical models, providing the basis for the formulation of an intentional account of decision-making. Although much progress has been made in human studies, a characterization of the neural underpinnings of an integrative mechanism, where evidence accumulation and the selection and execution of responses are carried out by the same system, remains challenging. Here, by employing magnetoencephalographic recording in combination with an experimental protocol that measures saccadic response and leverages a systematic modulation of evidence levels, we obtained a spectral dissociation between evidence accumulation mechanisms and motor preparation within the same brain region. Specifically, we show that within the dorsomedial parietal cortex alpha power modulation reflects the amount of sensory evidence available while beta power modulations reflect motor preparation, putatively representing the human homolog of the saccadic-related LIP region.

Looking through the windows: a study about the dependency of phase-coupling estimates on the data length.

OBJECTIVE: Being able to characterize functional connectivity (FC) state dynamics in a real-time setting, such as in brain-computer interface, neurofeedback or closed-loop neurostimulation frameworks, requires the rapid detection of the statistical dependencies that quantify FC in short windows of data. The aim of this study is to characterize, through extensive realistic simulations, the reliability of FC estimation as a function of the data length. In particular, we focused on FC as measured by phase-coupling (PC) of neuronal oscillations, one of the most functionally relevant neural coupling modes. APPROACH: We generated synthetic data corresponding to different scenarios by varying the data length, the signal-to-noise ratio, the phase difference value, the spectral analysis approach (Hilbert or Fourier) and the fractional bandwidth. We compared seven PC metrics, i.e. imaginary part of phase locking value (PLV), PLV of orthogonalized signals, phase lag index (PLI), debiased weighted PLI, imaginary part of coherency, coherence of orthogonalized signals and lagged coherence. MAIN RESULTS: Our findings show that, for a signal-to-noise-ratio of at least 10 dB, a data window that contains 5 to 8 cycles of the oscillation of interest (e.g. a 500-800ms window at 10Hz) is generally required to achieve reliable PC estimates. In general, Hilbert-based approaches were associated with higher performance than Fourier-based approaches. Furthermore, the results suggest that, when the analysis is performed in a narrow frequency range, a larger window is required. SIGNIFICANCE: The achieved results pave the way to the introduction of best-practice guidelines to be followed when a real-time frequency-specific PC assessment is at target.