The dynamics between coupled brains of individuals have been increasingly represented by inter-brain synchronization (IBS) when they coordinate with each other, mostly using simultaneous-recording signals of brains (namely hyperscanning) with fNIRS. In fNIRS hyperscanning studies, IBS has been commonly assessed through the wavelet transform coherence (WTC) method because of its advantage on expanding time series into time-frequency space where oscillations can be seen in a highly intuitive way. The observed IBS can be further validated via the permutation-based random pairing of the trial, partner, and condition. Here, a protocol is presented to describe how to obtain brain signals via fNIRS technology, calculate IBS through the WTC method, and validate IBS by permutation in a hyperscanning study. Further, we discuss the critical issues when using the above methods, including the choice of fNIRS signals, methods of data preprocessing, and optional parameters of computations. In summary, using the WTC method and permutation is a potentially standard pipeline for analyzing IBS in fNIRS hyperscanning studies, contributing to both the reproducibility and reliability of IBS.
The dynamics between coupled brains of individuals have been increasingly represented by inter-brain synchronization (IBS) when they coordinate with each other, mostly using simultaneous-recording signals of brains (namely hyperscanning) with fNIRS. In fNIRS hyperscanning studies, IBS has been commonly assessed through the wavelet transform coherence (WTC) method because of its advantage on expanding time series into time-frequency space where oscillations can be seen in a highly intuitive way. The observed IBS can be further validated via the permutation-based random pairing of the trial, partner, and condition. Here, a protocol is presented to describe how to obtain brain signals via fNIRS technology, calculate IBS through the WTC method, and validate IBS by permutation in a hyperscanning study. Further, we discuss the critical issues when using the above methods, including the choice of fNIRS signals, methods of data preprocessing, and optional parameters of computations. In summary, using the WTC method and permutation is a potentially standard pipeline for analyzing IBS in fNIRS hyperscanning studies, contributing to both the reproducibility and reliability of IBS.
When people coordinate with others, their brains and bodies become a coupled unit through continuous mutual adaption. The coupling between brains can be represented by inter-brain synchronization (IBS) through the hyperscanning approach, which simultaneously records two or more individuals' brain signals1. Indeed, a growing body of fNIRS/EEG hyperscanning studies has found IBS in various collaboration contexts, including finger tapping2, group walking3, playing drums4, guitar playing5, and singing/humming6. fNIRS is widely used for the research of IBS during social interaction, as it less restricts head/body motions in relatively natural settings (compared to fMRI/EEG)7.
The article presents a protocol for calculating IBS via wavelet transform coherence (WTC) method in an fNIRS hyperscanning study. WTC is a method for assessing the cross-correlation between two movement signals on the time-frequency plane and, therefore, can give more information than the traditional correlation analysis (e.g., Pearson correlation and cross-correlation), which is only in the time domain8. In addition, hemodynamic signals are transformed into wavelet components, which can effectively remove the low-frequency noise. Although WTC is time-consuming, it has been the most commonly used method of calculating IBS in action imitation9, cooperative behavior10, verbal communication11, decision making12, and interactive learning13.
The article also presents how to validate IBS with the permutation-based random paring of trials, conditions, and participants. The IBS in hyperscanning studies is always proposed to track online social interaction between individuals, while it can also be interpreted by other explanations, such as the stimulus similarity, motion similarity, or condition similarity14. Permutation test, also called randomization test, can be leveraged to test the above-mentioned null hypotheses through resampling the observed data15. By using permutation, it is useful to investigate whether the identified IBS is specific to interactive behavior, ranging from modulation of IBS within dyads to between groups of partners16.
The protocol described here details how to obtain brain signals via fNIRS technology, calculate IBS through the WTC method, and validate IBS by permutation testing in a hyperscanning study. This study aims to examine whether privileged IBS is elicited by music meters during social coordination. The brain signals were recorded in the frontal cortex, based on the location of the IBS in a previous finding1. The experimental task was originally developed by Konvalinka and her colleges17, in which participants were asked to tap their fingers with the auditory feedback from the partner or themselves after listening to the meter or non-meter stimuli.
The protocol presented here was approved by the University Committee on Human Research Protection of East China Normal University.
1. Preparation for the experiment
2. Before participants arrive
NOTE: Ensure to follow steps 2.1-2.5 before participants arrive at the laboratory.
3. Participant arrival in the laboratory
NOTE: Sincerely appreciate the two participants of one dyad when they arrive at the fNIRS lab. Request them to put their phone on silent mode and temporarily leave their personal belongings in the cabinet. Then conduct the following processes in sequence:
4. Data analysis
NOTE: Perform all data analysis by using MATLAB software, with the following toolboxes: HOMER219, Hitachi2nirs20, xjView21, Cross Wavelet and Wavelet Coherence toolbox22, and Groppe's scripts in MathWork23.
The results showed that there was IBS at channel 5 in the meter coordination condition, whereas no IBS existed in other conditions (i.e., meter independence, non-meter coordination, non-meter independence; Figure 2A). At channel 5, the IBS in the meter coordination condition was significantly higher than the coherence values in the non-meter coordination and meter independence condition (Figure 2B). Channel 5 approximately belonged to the left dorsolateral prefrontal cortex (DLPFC; Brodmann Area 9). Moreover, the permutation analysis showed that the observed IBS probably presented in two individuals of one dyad who tried to synchronize with each other in the matched time, but not in the time, partner, or condition of randomly pairing (Figure 2C). Together, these results indicated that music meter induced privileged IBS at DLPFC during interpersonal coordination. Considering the role of DLPFC in social interaction (e.g., modulating attention to other persons28,29) and music (e.g., enhancing cognitive performance in the presence of a musical background30,31), the observed DLPFC-IBS in the meter coordination condition might be related to drive more attention resource to the process involved in interpersonal coordination, such as perceiving and understanding the partner's task and movement.
Figure 1: Experimental design. (A) Experimental procedure and task. (B) Probe configuration. (C) Experimental setup. Please click here to view a larger version of this figure.
Figure 2: Inter-brain synchronization (IBS). (A) The heat maps of the permutation test on the coherence value for each condition. There was IBS at channel 5 in the meter coordination condition. (B) The IBS at channel 5 in the meter coordination condition was significantly greater than those in the meter independence and non-meter coordination condition. **p < 0.01, *p < 0.05. Error bars represent minimum/maximum values. The diamond dots denote extreme values. The shaded area indicates the 95% confidence interval. (C) The effect of IBS (statistical z values) with permutating trial, individual, and condition for all channels. The dashed line indicates the effect of the IBS at channel 5 in the meter coordination condition. The x-axis represents the Z value, and the y-axis represents the number of samples. Please click here to view a larger version of this figure.
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This protocol provides a step-by-step procedure to calculate and validate IBS, using the fNIRS hyperscanning approach to simultaneously collect two participants' brain signals. Some critical issues involved in fNIRS data preprocessing, IBS calculation, statistics, and IBS validation are discussed below.
Data Preprocessing
It is necessary to preprocess fNIRS data in hyperscanning studies to extract real signals from the possible noise (i.e., motion artifacts, systemic components). Although the preprocess is skipped when analyzing IBS in earlier fNIRS hyperscanning studies10,32,33, it has been an essential and standard part in recent ones. In this study, both CBSI and PCA are used to remove noise; the former is reliable to remove head motion artifacts34, while the latter is good at decreasing the global physiological noise (e.g., respiratory, blood pressure, and blood flow variation)35. Of course, there are other motion correction methods for data preprocessing, which perform well in empirical fNIRS studies, such as wavelet filtering36, spline interpolation37, Kalman filtering38, autoregressive algorithms39, and short-channel separation correction40. The comparisons of motion correction methods reported that it is always better to correct motion artifacts than excluding channels or rejecting trials and that each method has emphasis particularly on. It has been proposed that adopting several motion correction methods simultaneously41, as shown in this study, is a realistic solution. In addition, low-pass and high-pass filtering are also usually used in fNIRS data preprocessing to remove physiological noise. Although this method is effective, it may destroy the task effect when the physiological noise and task effect occur in similar frequency bands42. Together, simultaneously using PCA and CBSI might be advisable for data preprocessing in fNIRS hyperscanning studies.
Calculate IBS
It has been proposed that more work is needed to standardize the IBS analysis steps and increase the reproducibility of IBS, as precise algorithms used to calculate IBS are variable across labs and studies43. In this work, the standard pipeline of calculating IBS through WTC is useful for researchers. There are several things needed to be careful. First, WTC commonly falls under the Morlet wavelet family, which is used in this study. However, it is proposed that a Complex Gaussian wavelet is more suitable for fNIRS data than a Morlet wavelet, as the former matches the waveform of the underlying signal (i.e., the multicycle signals rarely occur, especially for the signal of wavelengths around 10 to 20 s)44. More considerations should be directed to the wavelet coherence computations that affect the power of the analysis in subsequent applications for NIRS signals acquired during live social interactions. Second, to be consistent with previous findings of interpersonal coordination with music2,45,46 and music activities4,47,48, the coherence values were computed between the same channels in this study, while some studies have averaged the coherence values of all channels within the same brain region before statistical analysis49,50. In addition, the coherence values were calculated not only between the same channels/regions10,32,51 but also across different channels/regions52,53. These mentioned processes have enriched the pipeline of calculating IBS and might interest future directions of social interaction. Last but not least, only oxyHb values were analyzed in this study since oxyHb values are regarded as the most sensitive indicator of changes in the regional cerebral blood flow54. However, some researchers focused on deoxyHb changes, based on the findings that deoxyHb values are most closely related to the fMRI signal and independent of the global physiological noise55. Anyhow, the results might be more reliable if similar IBS effects are revealed in both oxyHb and deoxyHb changes. Therefore, the analysis of IBS on deoxyHb values is also necessary for future fNIRS hyperscanning studies.
Validate IBS
It is necessary to validate the revealed IBS, as the interpretation of IBS remains complex. For instance, IBS has been explained as a mechanism for information transmission, shared intentionality, behavioral alignment, similar perception, etc. It would help clarify the interpretation of IBS by performing null hypothesis testing with permutation, in which coherence values are either computed for the real dyads but randomly pairing trials or for fake dyads by randomly pairing participants within one condition/group or between conditions/groups16. In this study, permutation was performed by simply conducting a very large number of resamples (i.e., 1000 times). In contrast, coherence values can be calculated for all possible random pairs56. In addition, the above permutation test can be used to generate a null distribution of coherences from all possible coherences in the experiment, to see whether the observed IBS are near the top end of this distribution, which has been commonly used in studies that adopt real-life stimuli and experimental environment57,58. This analysis ensures that the IBS is real-interaction-specific at the sequence level, as the coherence values during matching ones (i.e., trials, individuals, and conditions) must on average statistically exceed an equal-sized random draw of coherences within or between dyads. Such a method is different from the baseline used in the current work (i.e., the resting-state coherence values), which is in line with traditional General Linear Models designs and is selected to compare the current results with the findings in previous studies. It should be noted that the 20-s-resting baseline in this study is shorter than the widely used duration (30 s or more than 1 min), which is used to restrict the total time of the experiment to 30 min to ensure the comfort of participants.
In conclusion, this article provides a specific pipeline of analyzing IBS in fNIRS hyperscanning studies. Such pipeline is a potentially standard data processing approach in the field, which will contribute to both the reproducibility and reliability of IBS. In the future, the details of data processing should be further refined when analyzing IBS for particular groups (i.e., parent-infant, children, and schizophrenia patients) and particular contexts (i.e., nonverbal or verbal communication and teaching situations). Finally, showcasing the protocol of analyzing the inter-brain network for larger groups of participants in natural interactions will benefit the quantification of social interaction.
The authors have nothing to disclose.
This research was supported by: National Natural Science Foundation of China (31872783, 31800951).
Computer | Hewlett-Packard Development Company, L.P. | HP S01-pF157mcn | |
Earphone | Royal Philips Electronics, Eindhoven, The Netherlands | SHE2405BK/00 | |
EEG cap | Compumedics Neuroscan, Charlotte, USA | 64-channel Quik-Cap | |
E-Prime software | Psychology Software Tools, Inc., Pittsburgh, USA | E-Prime 3 | |
fNIRS system | Hitachi Medical Corporation, Tokyo, Japan | ETG-7100 Optical Topography System | |
MATLAB 2014b | The MathWorks, Inc., Natick, MA | MATLAB 2014b | |
MuseScore | Musescore Company, Belgium | MuseScore 3.6.2.548021803 | |
Swimming cap | Decathlon Group, Villeneuve-d'Ascq, France | 1681552 |