The author would like to thank his many thesis students and research assistants for their help in piloting and administering this protocol. Data presented in this manuscript is based upon work supported by the National Science Foundation (NSF) under Grant Number (1147743) to Jason Themanson and a grant from Illinois Wesleyan University to Jason Themanson.

NOTE: The following protocol was developed in accordance with ethical standards approved by the Institutional Review Board at Illinois Wesleyan University.
1. Cyberball Stimulus Preparation
<ol>
	<li>Download the Cyberball paradigm12,18 and install it on the computer (the current protocol utilized images from Cyberball version 3.0). Alternatively, create computerized images to recreate the Cyberball paradigm to meet specific needs.</li>
	<li>Create individual images for each portion of the throws within Cyberball by using a photo-editing program. For example, break down each of the throws from player to player into the individual throw frames that are shown one after another to create the image of a ball being thrown from player to player on the computer screen (see Figure 1).</li>
	<li>Add any labels, names, or pictures to each individual throw frame in the photo-editing program, including anything to represent the human participant as the bottom player on the screen (represented by the hand at the bottom of the screen in Figure 1) to create a series of throw frames that are identical except for the movement of the ball from player to player.</li>
	<li>Note which frame in each throw sequence is the &#8220;informational frame&#8221; for that throw, or the first frame within the throw sequences that provides information to the players about the specific destination of the throw (i.e., which other player will receive the ball).</li>
	<li>Ensure that there are throw sequences creating a throw from each player to each other player on the screen (including throws from the human participant to the other players), that each throw sequence has the same number of throw frames, and the informational frame within each throw sequence has been noted.</li>
</ol>
2. Cyberball Social Interaction Programming
<ol>
	<li>Create a sequence file using stimulus presentation software to detail the exact sequence of events within the Cyberball social interaction.
	<ol>
		<li>For the sequence file, specify the specific throw frames (in order), the timing of the frames on the screen, the sequencing of the frames, the nature of the event (throw from whom to whom), the response required by the human participant (when necessary), and the overall order of events to create the desired interaction. Explicitly enter all of these specifications into the proper rows, columns, and spaces within the programming code during the creation of the sequence file.</li>
		<li>Specify all of the above-mentioned specifics within the programming code for each event within the sequence file and repeat the steps for each sequence file created (e.g., inclusion, exclusion).
		<ol>
			<li>Order each of the throw frames in the correct sequence within the sequence file so that the first ball toss is completed without error from one player to the other. Create similar ordered sequences in the file for each type of throw among the players so that each type of throw is represented in the sequence file (e.g., a three-player game consists of six different possible throws).</li>
			<li>Space the timing of each throw frame 450 msec apart. In this method, ensure that each frame appears for 450 msec before being replaced by the next frame, which provides an image of motion on the screen for the participant and creates a throw event that lasts a total of 2,700 msec.</li>
			<li>Insert an event-related marker each time an informational frame is presented in the sequence file so that the presentation of the informational frame can be marked in time in the file saving the participants&#8217; neural activity. Code this marker to represent the nature of the event by using numbers to represent the players (left player is player &#8220;1,&#8221; the bottom player is player &#8220;2,&#8221; the right player is player &#8220;3&#8221;), which would allow the code &#8220;13&#8221; to represent a throw from the player on the left to the player on the right.</li>
			<li>Copy the entire set of six different throw sequences within the file so that each throw sequence is represented at least twice within the sequence file. This will provide programming flexibility to change the order of events within each block so that they do not look pre-determined.</li>
			<li>Create &#8220;if, then&#8221; statements within the sequence file to allow the human participant to freely select which player will receive the next throw following the human participant. Give the human participant a response pad or mouse to select the next action after receiving a ball toss; potentially using the right mouse button to throw to the player on the right and using the left mouse button to throw to the player on the left in a three-player game.</li>
			<li>Ensure that the &#8220;if, then&#8221; statements lead to the appropriate next throw sequence so that the game play appears seamless (i.e., a human throw to the player on the left should be followed by a throw from the player on the left to another player).</li>
			<li>Create loops and &#8220;if, then&#8221; statements within the sequence file to represent the desired game action and allow the program to appropriately move to the next event regardless of the selections of the human participant.</li>
			<li>Initiate counters within the program to change the nature of the game so that the program does not become apparent to the human player (i.e., the same computerized player does not always make the same throw). Use these counters to switch the game action and remove patterns of play throughout the game after the repeated occurrence of a specific event or pattern of events to better give the appearance of spontaneous live play among players on the sides of the screen, not just the actual human participant represented on the bottom of the screen.</li>
		</ol>
		</li>
	</ol>
	</li>
	<li>Develop different sequence files in order to study different types of social interactions. Make these interactions largely inclusive or exclusive, or even partially inclusive or exclusive, for the human participant depending on the nature of the research question by varying the proportion and order of inclusionary events and exclusionary events within each sequence file.</li>
	<li>Ensure that the event markers appear in the EEG files when collecting neural data to create event-related brain potentials (ERPs) for each event type within each of the different social interactions. These markers should appear in the EEG file as the informational frame is presented to the participant.</li>
</ol>
3. Neuroelectric Recording
<ol>
	<li>Prepare participants for electroencephalography (EEG) assessment in accordance with the guidelines of the Society for Psychophysiological Research19.</li>
	<li>Use a lycra electrode cap embedded with 64 sintered Ag-AgCl electrodes (10 mm), arranged in a 10&#8211;10 system montage20 to collect EEG data. Fit the cap on the participant&#8217;s head and prepare each electrode using conductive gel.
	<ol>
		<li>Reference the electrodes online to an electrode placed at the midpoint between Cz and CPz and use AFz as the ground electrode. 
		NOTE: Alternative online references may be needed depending upon the nature of the electrode cap used for data collection.</li>
		<li>Collect vertical and horizontal bipolar electrooculographic activity (EOG) to monitor eye movements using sintered Ag-AgCl electrodes placed above and below the right orbit and near the outer canthus of each eye.</li>
	</ol>
	</li>
	<li>Use a digital bioamplifier to continuously digitize (500 Hz sampling rate), amplify (gain depends on the specific amplifier), and filter (70 Hz low-pass filter, including a 60 Hz notch filter) the raw EEG signal in DC mode. Choose these setting from the available options in the EEG analysis software for the amplifier prior to data collection and vary depending on the specifications of the EEG hardware and software.</li>
	<li>Record EEG activity using EEG analysis software in order to further process the neural data.</li>
</ol>
4. Offline Neuroelectric Data Processing
<ol>
	<li>Correct eye blinks using a spatial filter, a multi-step procedure that generates an average eye blink, utilizes a spatial singular value decomposition based on principal component analysis (PCA) to extract the first component and covariance values, and then uses those covariance values to develop a filter that is specifically sensitive to eye blinks21.</li>
	<li>Create stimulus-locked epochs relative to the event marker that was inserted in the continuous EEG file in the EEG analysis software by selecting this function from the choices of data transformation options. Run these epochs from -900 msec to 1,800 msec relative to inserted marker, which is equivalent to the entire duration of each six-frame throw and has a timepoint of 0 msec where the event marker was inserted as displayed in Figure 1.</li>
	<li>Correct for baseline difference between the epochs by removing the average pre-stimulus baseline activity from each epoch (i.e., the 900 msec time window that runs from -900 msec to 0 msec prior to the event marker). This function can be selected or initiated from the data transformation options available in the EEG analysis software.</li>
	<li>Low-pass filter (30 Hz; 24 dB/octave) the epochs and reject any epochs with electrical artifacts that exceed +75V. Choose these setting from the available options in the EEG analysis software allowing the transformation of EEG data following data collection and vary depending on the specifications of the EEG software.</li>
	<li>Average the neural responses together for each event type within the Cyberball task blocks. 
	NOTE: This averaging process can be adapted to only average the first 20, middle 20, or even last 20 events of a similar type within each interaction to examine dynamic patterns of neural activation over the course of the social interaction13.
	<ol>
		<li>Combine the various events types to create three major event categories: throws to the participant from either other player, throws from the participant to either other player, and throws not including the participant between the two other players. For example, combine throws from the human to the left player and to the right player into one average waveform.</li>
		<li>Combine the events from the computerized players into the event types of most interest: throws to the human participant (inclusionary) and throws away from the human participant (exclusionary).</li>
	</ol>
	</li>
	<li>If applicable, quantify the N2 component as the average amplitude in the discrete latency window running from 200 - 320 msec after the event marker at FCz.</li>
	<li>If applicable, quantify the P3 component as the average amplitude in the discrete latency window running from 320 - 450 msec following the event marker at Pz.</li>
	<li>If applicable, quantify ERP components to throw frames following the informational frame to examine ongoing differences among patterns of neural activity to different event types with the social interaction.</li>
</ol>

<ol>
	<li>Baumeister, R. F., DeWall, C. N., Ciarocco, N. J., Twenge, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Social+exclusion+impairs+self-regulation.">Social exclusion impairs self-regulation.</a> J Pers Soc Psychol. 88, 589-604 (2005).</li><li>Baumeister, R. F., Twenge, J. M., Nuss, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+social+exclusion+on+cognitive+processes:+Anticipated+aloneness+reduces+intelligent+thought.">Effects of social exclusion on cognitive processes: Anticipated aloneness reduces intelligent thought.</a> J Pers Soc Psychol. 83, 817-827 (2002).</li><li>Eisenberger, N. I., Lieberman, M. D., Williams, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Does+rejection+hurt:+An+fMRI+study+in+social+exclusion.">Does rejection hurt: An fMRI study in social exclusion.</a> Science. 302, 290-292 (2003).</li><li>Masten, C. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+correlates+of+social+exclusion+during+adolescence:+understanding+the+distress+of+peer+rejection.">Neural correlates of social exclusion during adolescence: understanding the distress of peer rejection.</a> Soc Cogn Affect Neur. 4, 143-157 (2009).</li><li>Williams, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Ostracism: The power of silence. , (2001).</li><li>Williams, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ostracism.">Ostracism.</a> Annu Rev Psychol. 58, 425-452 (2007).</li><li>Pickett, C. L., Gardner, W. L., Williams, D. K., Forgas, P. J., von Hippel, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+social+monitoring+system:+Enhanced+sensitivity+to+social+cues+as+an+adaptive+response+to+social+exclusion.">The social monitoring system: Enhanced sensitivity to social cues as an adaptive response to social exclusion.</a> The Social Outcast: Ostracism, Social Exclusion, Rejection, and Bullying. , 213-226 (2005).</li><li>Richman, L. S., Leary, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reactions+to+discrimination,+stigmatization,+ostracism,+and+other+forms+of+interpersonal+rejection:+A+multimotive+model.">Reactions to discrimination, stigmatization, ostracism, and other forms of interpersonal rejection: A multimotive model.</a> Psychol Rev. 116, 365-383 (2009).</li><li>Wesselmann, E. D., Wirth, J. H., Mroczek, D. K., Williams, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dial+a+feeling:+Detecting+moderation+of+affect+decline+during+ostracism.">Dial a feeling: Detecting moderation of affect decline during ostracism.</a> Pers Individ Dif. 53, 580-586 (2012).</li><li>Eisenberger, N. I., Lieberman, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Why+rejection+hurts:+a+common+neural+alarm+system+for+physical+and+social+pain.">Why rejection hurts: a common neural alarm system for physical and social pain.</a> Trends Cogn Sci. 7, 294-300 (2004).</li><li>Eisenberger, N. I., Gable, S. L., Lieberman, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=fMRI+responses+relate+to+differences+in+real-world+social+experience.">fMRI responses relate to differences in real-world social experience.</a> Emotion. 7, 745-754 (2007).</li><li>Williams, K. D., Cheung, C. K. T., Choi, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cyberostracism:+Effects+of+being+ignored+over+the+internet.">Cyberostracism: Effects of being ignored over the internet.</a> J Pers Soc Psychol. 79, 748-762 (2000).</li><li>Themanson, J. R., Khatcherian, S. M., Ball, A. B., Rosen, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+event-related+examination+of+neural+activity+during+social+interactions.">An event-related examination of neural activity during social interactions.</a> Soc Cogn Affect Neur. 8, 727-733 (2013).</li><li>Coles, M. G. H., Gratton, G., Fabinani, M., Cacioppo, J. T., Tassinary, L. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Event-related+brain+potentials.">Event-related brain potentials.</a> Principals of Psychophysiology: Physical, Social and Inferential Elements. , 413-455 (1990).</li><li>Crowley, M. J., Wu, J., Molfese, P. J., Mayes, L. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Social+exclusion+in+middle+childhood:+Rejection+events,+slow-wave+neural+activity,+and+ostracism+distress.">Social exclusion in middle childhood: Rejection events, slow-wave neural activity, and ostracism distress.</a> Soc Neurosci. 5, 483-495 (2010).</li><li>Gutz, L., K&#252;pper, C., Renneberg, B., Niedeggen, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Processing+social+participation:+an+event-related+brain+potential+study.">Processing social participation: an event-related brain potential study.</a> NeuroReport. 22, 453-458 (2011).</li><li>Themanson, J. R., Ball, A. B., Khatcherian, S. M., Rosen, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+social+exclusion+on+the+ERN+and+the+cognitive+control+of+action+monitoring.">The effects of social exclusion on the ERN and the cognitive control of action monitoring.</a> Psychophysiology. 51, 215-225 (2014).</li><li>Williams, K. S., Yeager, D. S., Cheung, C. K. T., Choi, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Cyberball (version 4.0) [Software]. , (2012).</li><li>Picton, T. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Guidelines+for+using+human+event-related-potentials+to+study+cognition:+Recording+standards+and+publication+criteria.">Guidelines for using human event-related-potentials to study cognition: Recording standards and publication criteria.</a> Psychophysiology. 37, 127-152 (2000).</li><li>Chatrian, G. E., Lettich, E., Nelson, P. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ten+percent+electrode+system+for+topographic+studies+of+spontaneous+and+evoked+EEG+activity.">Ten percent electrode system for topographic studies of spontaneous and evoked EEG activity.</a> Am J EEG Technol. 25, 83-92 (1985).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Offline Analysis of Acquired Data SCAN 4.3 &#8211;Vol. II, EDIT 4.3). [Software Manual]. , (2003).</li><li>Crowley, M. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exclusion+and+micro-rejection:+Event-related+potential+response+predicts+mitigated+distress.">Exclusion and micro-rejection: Event-related potential response predicts mitigated distress.</a> NeuroReport. 20, 1518-1522 (2009).</li><li>Gardner, W. L., Pickett, C. L., Brewer, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Social+exclusion+and+selective+memory:+How+the+need+to+belong+influences+memory+for+social+events.">Social exclusion and selective memory: How the need to belong influences memory for social events.</a> Pers Soc Psychol Bull. 26, 486-496 (2000).</li><li>Smith, A., Williams, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=R+U+there?+Ostracism+by+cell+phone+text+message.">R U there? Ostracism by cell phone text message.</a> Group Dyn: Theory Res Pract. 8, 291-301 (2004).</li><li>Luck, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> An Introduction to the Event-Related Potential Technique. , (2005).</li></ol>

The author has nothing to disclose and declares no conflict of interest.

In this article, a protocol allowing for the measurement of ongoing event-related neural and behavioral data during social interactions was presented. This procedure creates opportunities to look at multiple different event types (inclusionary, exclusionary) within and across varied social interactions. Specifically, the procedure can quantify moment-to-moment event-related neural activity in response to any event that occurs during a computerized social interaction. This neural activity is independent of any particular ...

The scientific examination of social interactions has undergone a renaissance in recent years, with an explosion of new theoretical explanations, models, and paradigms aimed at understanding and exploring the effects of being the target or source of social exclusion and how those interactions lead to the many consequences of exclusion1-6. Though the literature had made tremendous strides in developing a better understanding of the consequences of social exclusion at behavioral, emotional, cognitive, and neural levels, a great deal remains unknown in relation to the dynamics involved in social exclusion. One notable gap in the literature relates to the measurement of various dynamic social exclusion processes during social interactions. For instance, multiple theoretical models3,5-8 suggest that the monitoring and assessment of instances of social exclusion is an initial step in a larger self-regulatory system aimed at coping with social exclusion and maintaining healthy and acceptable levels of belonging and social inclusion. These models, and much of the existing literature on exclusion, provide tremendous insights into the consequences of social exclusion and the harmful effects exclusion causes on neural, behavioral, cognitive, and emotional levels. However, the specific processes ongoing in targets of exclusion during social interactions, which lead to both the perception of exclusion and the subsequent emotional and cognitive reactions to exclusion, remain undefined. Researchers have adapted methodologies to obtain self-reported feeling states during social interactions9, but these data did not examine the ongoing neural processes that may motivate any self-reported effects.
Accordingly, examinations of exclusion during social interactions were initiated using functional magnetic resonance imaging (fMRI) to &#8220;see&#8221; what is happening while individuals are being excluded3,4,10,11. These studies revealed different patterns of neural activation during exclusion compared to inclusion. Though tremendously important in enhancing the understanding of ongoing neural processes present during exclusion and their relations with the self-reported consequences of being excluded, these studies are limited in how they can represent the dynamic nature of social interactions. Specifically, these fMRI methodologies aggregated neural activity across entire social interactions and were unable to examine exclusion on a moment-to-moment basis. This limitation prohibits a complete understanding of the dynamic nature of exclusion-related emotional and cognitive processing that is taking place during social interactions as researchers are unable to determine which moments or events during an exchange are meaningful in relation to the development of one&#8217;s perceptions of exclusion and the associated emotional response.
To address these limitations, recent research has implemented the measurement of one class of neural activity, known as event-related brain potentials (ERPs), during the execution of the Cyberball paradigm12 to examine the moment-to-moment patterns of neural activation present during social exclusion13. ERPs refer to neuroelectric activity measured on the scalp that is time-locked to discrete events and represents brain activity in response to or in preparation for a stimulus or response14. Further, ERPs possess a superior temporal resolution when compared to fMRI, which provides valuable insights into the dynamic responses to social exclusion. As such, neural indices obtained through the event-related examination of brain activity in response to instances of social inclusion and exclusion, which can be implemented and controlled through the Cyberball paradigm and are described in the present protocol, are necessary to evaluate the models and predictions present in current social exclusion theory.
The goal of the current methodology is to measure ongoing neural responses to social events (inclusionary events, exclusionary events) during computerized social interactions in a human participant. In this methodology, researchers have the ability to quantify neural activity in response to each event within the interaction. Further, the current protocol allows for the ongoing examination of each social event as each event is made up of multiple throw images. This allows researchers to look at changes in neural activity as the events unfold. This level of analysis is not available in other methodologies that examine ERPs during social interactions15,16 as these methodologies only capture neural activity in relation to one image for each event without allowing for the examination of the unfolding event as it occurs. Additionally, the human participant is led to believe that he or she is playing an online game with other people, but is actually playing within a pre-programmed game with a computer. Because the interaction is pre-programmed within the computer, with the flexibility to interact with the decisions made by the human participant, the nature of the social interactions can be pre-determined and programmed to vary depending on the nature of the research question13,17. For example, the behavior of the computer-generated players during the protocol can be tailored to create instances of social inclusion or social exclusion of any specified duration by altering the pre-programmed schedule of throws (e.g., which player throws the ball to which other player, when those throws occur, the number of throws, and the timing of throws). Accordingly, this allows researchers to measure neural activity in response to events that may or may not match the overall context of the interaction. For example, researchers can quantify a participant&#8217;s neural response to an exclusionary social event within an interaction that is largely inclusionary for that participant and potentially compare it to that participant&#8217;s neural response to an exclusionary event within a largely exclusionary interaction. These research opportunities are not readily available using fMRI technology given the temporal limitations of fMRI. With this programming flexibility, the current protocol allows researchers from various neuroscientific and psychological backgrounds to address research questions in new ways and obtain dynamic neural and behavioral activity during social interactions.

<table border="0" cellpadding="0" cellspacing="0" width="720">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Name of Material/ Equipment</td>
			<td style="width:219px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Cyberball (Williams et al., 2000) computerized social interaction program</td>
			<td>https://cyberball.wikispaces.com</td>
			<td style="width:119px;"></td>
			<td>An Alternate set of computerized images can be used or created by the researcher</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Neuroscan SynAmps2 64-Channel Amplifier with SCAN 4.3.1 Acquisition and Analysis Software</td>
			<td style="width:219px;">Compumedics Neuroscan Neuromedical Supplies - http://www.neuroscan.com/supplies.cfm</td>
			<td style="width:119px;">9032-0010-01</td>
			<td>Alternate amplifiers and EEG acquisition equipment and sofware can be used</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">STIM2 Complete Version 2.1 Stimulus Presentation Software</td>
			<td style="width:219px;">Compumedics Neuroscan Neuromedical Supplies - http://www.neuroscan.com/supplies.cfm</td>
			<td style="width:119px;">666M</td>
			<td>Alternate stimulus presenation software can be used</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">SynAmps2 Quik-Cap Sintered Ag/AgCl 64 Ch./Medium</td>
			<td style="width:219px;">Compumedics Neuroscan Neuromedical Supplies - http://www.neuroscan.com/supplies.cfm</td>
			<td style="width:119px;">96050255</td>
			<td>Alternate EEG caps can be used</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Quik-Gel Conductive Gel</td>
			<td style="width:219px;">Compumedics Neuroscan Neuromedical Supplies - http://www.neuroscan.com/supplies.cfm</td>
			<td style="width:119px;">92000016</td>
			<td>Alternate EEG conductive electrode gel can be used</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">NuPrep</td>
			<td style="width:219px;">Compumedics Neuroscan Neuromedical Supplies - http://www.neuroscan.com/supplies.cfm</td>
			<td style="width:119px;">92100025</td>
			<td>Alternate skin preparation exfoliants can be used</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Blunt needle and syringe kit</td>
			<td style="width:219px;">Compumedics Neuroscan Neuromedical Supplies - http://www.neuroscan.com/supplies.cfm</td>
			<td style="width:119px;">104207</td>
			<td>Needle and syringe kit is used to apply conductive gel to electrode embedded in the EEG cap</td>
		</tr>
	</tbody>
</table>

measuring neural and behavioral activity during ongoing computerized social interactions: an examination of event-related brain potentials

Social exclusion is a complex social phenomenon with powerful negative consequences. Given the impact of social exclusion on mental and emotional health, an understanding of how perceptions of social exclusion develop over the course of a social interaction is important for advancing treatments aimed at lessening the harmful costs of being excluded. To date, most scientific examinations of social exclusion have looked at exclusion after a social interaction has been completed. While this has been very helpful in developing an understanding of what happens to a person following exclusion, it has not helped to clarify the moment-to-moment dynamics of the process of social exclusion. Accordingly, the current protocol was developed to obtain an improved understanding of social exclusion by examining the patterns of event-related brain activation that are present during social interactions. This protocol allows greater precision and sensitivity in detailing the social processes that lead people to feel as though they have been excluded from a social interaction. Importantly, the current protocol can be adapted to include research projects that vary the nature of exclusionary social interactions by altering how frequently participants are included, how long the periods of exclusion will last in each interaction, and when exclusion will take place during the social interactions. Further, the current protocol can be used to examine variables and constructs beyond those related to social exclusion. This capability to address a variety of applications across psychology by obtaining both neural and behavioral data during ongoing social interactions suggests the present protocol could be at the core of a developing area of scientific inquiry related to social interactions.

This protocol has been used in previously published research examining the influence of social exclusion on ongoing neural and behavioral activity13. Twenty-two college-aged participants (15 females, 7 males) completed three sessions of the Cyberball task under conditions described above. After providing informed consent, participants were told that they would be playing a computerized ball-tossing game with other undergraduate participants. However, the other participants were not real, they were represented ...

Watch this Scientific Journal Video about Measuring Neural and Behavioral Activity During Ongoing Computerized Social Interactions: An Examination of Event-Related Brain Potentials at JoVE.com

Measuring Neural and Behavioral Activity During Ongoing Computerized Social Interactions: An Examination of Event-Related Brain Potentials

Research on social exclusion has grown tremendously in recent years. As the field expands, it is imperative to develop sophisticated methodologies allowing for the simultaneous measurement of neural and behavioral outcomes during social exclusion. This protocol utilizes event-related brain potentials to record ongoing neural activity during computerized social interactions.

Social exclusion is a complex social phenomenon with powerful negative consequences. Given the impact of social exclusion on mental and emotional health, ...

measuring-neural-behavioral-activity-during-ongoing-computerized

Research

Education

JoVE Journal

JoVE Core

Social Psychology

Behavior

Foundations of Social Psychology

Helping and Hurting

SCIENTISTS IN ACTION

13.6K Views.  Illinois Wesleyan University. Research on social exclusion has grown tremendously in recent years. As the field expands, it is imperative to develop sophisticated methodologies allowing for the simultaneous measurement of neural and behavioral outcomes during social exclusion. This protocol utilizes event-related brain potentials to record ongoing neural activity during computerized social interactions. 

Video: Measuring Neural and Behavioral Activity During Ongoing Computerized Social Interactions: An Examination of Event-Related Brain Potentials

Investigating the Effects of Antipsychotics and Schizotypy on the N400 Using Event-Related Potentials and Semantic Categorization

Within the field of cognitive neuroscience, functional magnetic resonance imaging (fMRI) is a popular method of visualizing brain function. This is in part because of its excellent spatial resolution, which allows researchers to identify brain areas associated with specific cognitive processes. However, in the quest to localize brain functions, it is relevant to note that many cognitive, sensory, and motor processes have temporal distinctions that are imperative to capture, an aspect that is left unfulfilled by fMRI&#8217;s suboptimal temporal resolution. To better understand cognitive processes, it is thus advantageous to utilize event-related potential (ERP) recording as a method of gathering information about the brain. Some of its advantages include its fantastic temporal resolution, which gives researchers the ability to follow the activity of the brain down to the millisecond. It also directly indexes both excitatory and inhibitory post-synaptic potentials by which most brain computations are performed. This sits in contrast to fMRI, which captures an index of metabolic activity. Further, the non-invasive ERP method does not require a contrast condition: raw ERPs can be examined for just one experimental condition, a distinction from fMRI where control conditions must be subtracted from the experimental condition, leading to uncertainty in associating observations with experimental or contrast conditions. While it is limited by its poor spatial and subcortical activity resolution, ERP recordings&#8217; utility, relative cost-effectiveness, and associated advantages offer strong rationale for its use in cognitive neuroscience to track rapid temporal changes in neural activity. In an effort to foster increase in its use as a research imaging method, and to ensure proper and accurate data collection, the present article will outline &#8211; in the framework of a paradigm using semantic categorization to examine the effects of antipsychotics and schizotypy on the N400 &#8211; the procedure and key aspects associated with ERP data acquisition.

Using event-related EEG potentials (ERPs), we investigate the effects of antipsychotic medications on abnormal semantic brain activations in healthy individuals with schizotypal traits. We use ERPs to track distinct changes in brain activity, shedding insight into the cognitive processes associated with semantic categorization.

Within the field of cognitive neuroscience, functional magnetic resonance imaging (fMRI) is a popular method of visualizing brain function. This is in ...

Infant Auditory Processing and Event-related Brain Oscillations

Rapid auditory processing and acoustic change detection abilities play a critical role in allowing human infants to efficiently process the fine spectral and temporal changes that are characteristic of human language. These abilities lay the foundation for effective language acquisition; allowing infants to hone in on the sounds of their native language. Invasive procedures in animals and scalp-recorded potentials from human adults suggest that simultaneous, rhythmic activity (oscillations) between and within brain regions are fundamental to sensory development; determining the resolution with which incoming stimuli are parsed. At this time, little is known about oscillatory dynamics in human infant development. However, animal neurophysiology and adult EEG data provide the basis for a strong hypothesis that rapid auditory processing in infants is mediated by oscillatory synchrony in discrete frequency bands. In order to investigate this, 128-channel, high-density EEG responses of 4-month old infants to frequency change in tone pairs, presented in two rate conditions (Rapid: 70 msec ISI and Control: 300 msec ISI) were examined. To determine the frequency band and magnitude of activity, auditory evoked response averages were first co-registered with age-appropriate brain templates. Next, the principal components of the response were identified and localized using a two-dipole model of brain activity. Single-trial analysis of oscillatory power showed a robust index of frequency change processing in bursts of Theta band (3 - 8 Hz) activity in both right and left auditory cortices, with left activation more prominent in the Rapid condition. These methods have produced data that are not only some of the first reported evoked oscillations analyses in infants, but are also, importantly, the product of a well-established method of recording and analyzing clean, meticulously collected, infant EEG and ERPs. In this article, we describe our method for infant EEG net application, recording, dynamic brain response analysis, and representative results.

High-density electroencephalography (dEEG) is being used increasingly to study brain development and plasticity in the early years of life. Here we present an application of sophisticated analysis techniques that builds on traditional EEG recording to understand the oscillatory dynamics of rapid auditory processing in the infant brain.

Rapid auditory processing and acoustic change detection abilities play a critical role in allowing human infants to efficiently process the fine spectral ...

The Modified Hole Board - Measuring Behavior, Cognition and Social Interaction in Mice and Rats

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure multiple dimensions of unconditioned behavior in small laboratory mammals (e.g., mice, rats, tree shrews and small primates). This paradigm is a valuable alternative for the use of a behavioral test battery, since a broad behavioral spectrum of an animal&#8217;s behavioral profile can be investigated in one single test.
The apparatus consists of a box, representing the &#8216;protected&#8217; area, separated from a group compartment. A board, on which small cylinders are staggered in three lines, is placed in the center of the box, representing the &#8216;unprotected&#8217; area of the set-up. The cognitive abilities of the animals can be measured by baiting some cylinders on the board and measuring the working and reference memory. Other unconditioned behavior, such as activity-related-, anxiety-related- and social behavior, can be observed using this paradigm. Behavioral flexibility and the ability to habituate to a novel environment can additionally be observed by subjecting the animals to multiple trials in the mHB, revealing insight into the animals&#8217; adaptive capacities.
Due to testing order effects in a behavioral test battery, na&#239;ve animals should be used for each individual experiment. By testing multiple behavioral dimensions in a single paradigm and thereby circumventing this issue, the number of experimental animals used is reduced. Furthermore, by avoiding social isolation during testing and without the need to food deprive the animals, the mHB represents a behavioral test system, inducing if any, very low amount of stress.

This protocol describes the modified hole board, which is a behavioral test set-up that comprises the characteristics of an open field and a traditional hole board. This set-up enables the differential analysis of unconditioned behavior of small laboratory mammals as well as the analysis of cognitive abilities.

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure ...

A Method for Remotely Silencing Neural Activity in Rodents During Discrete Phases of Learning

This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while animals are engaged in learning and memory tasks. The approach combines pharmacogenetics (Designer-Receptors-Exclusively-Activated-by-Designer-Drugs) with a behavioral paradigm (sensory preconditioning) that is designed to distinguish between different forms of learning. Specifically, viral-mediated delivery is used to express a genetically modified inhibitory G-protein coupled receptor (the Designer Receptor) into a discrete brain region in the rodent. Three weeks later, when designer receptor expression levels are high, a pharmacological agent (the Designer Drug) is administered systemically 30 min prior to a specific behavioral session. The drug has affinity for the designer receptor and thus results in inhibition of neurons that express the designer receptor, but is otherwise biologically inert. The brain region remains silenced for 2-5 hr (depending on the dose and route of administration). Upon completion of the behavioral paradigm, brain tissue is assessed for correct placement and receptor expression. This approach is particularly useful for determining the contribution of individual brain regions to specific components of behavior and can be used across any number of behavioral paradigms.

This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while rats are engaged in learning and memory tasks. The approach combines pharmacogenetics (Designer-Receptors-Exclusively-Activated-by-Designer-Drugs) with a behavioral paradigm (sensory preconditioning) that is designed to distinguish between different components of learning.

This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while animals are engaged in learning and ...

A Novel Experimental and Analytical Approach to the Multimodal Neural Decoding of Intent During Social Interaction in Freely-behaving Human Infants

Understanding typical and atypical development remains one of the fundamental questions in developmental human neuroscience. Traditionally, experimental paradigms and analysis tools have been limited to constrained laboratory tasks and contexts due to technical limitations imposed by the available set of measuring and analysis techniques and the age of the subjects. These limitations severely limit the study of developmental neural dynamics and associated neural networks engaged in cognition, perception and action in infants performing &#8220;in action and in context&#8221;. This protocol presents a novel approach to study infants and young children as they freely organize their own behavior, and its consequences in a complex, partly unpredictable and highly dynamic environment. The proposed methodology integrates synchronized high-density active scalp electroencephalography (EEG), inertial measurement units (IMUs), video recording and behavioral analysis to capture brain activity and movement non-invasively in freely-behaving infants. This setup allows for the study of neural network dynamics in the developing brain, in action and context, as these networks are recruited during goal-oriented, exploration and social interaction tasks.

This protocol presents a novel methodology for the neural decoding of intent from freely-behaving infants during unscripted social interaction with an actor. Neural activity is acquired using non-invasive high-density active scalp electroencephalography (EEG). Kinematic data is collected with inertial measurement units and supplemented with synchronized video recording.

Understanding typical and atypical development remains one of the fundamental questions in developmental human neuroscience. Traditionally, experimental ...

Event-related Potentials During Target-response Tasks to Study Cognitive Processes of Upper Limb Use in Children with Unilateral Cerebral Palsy

Unilateral Cerebral Palsy (CP) is a neurodevelopmental disorder that is a very common cause of disability in childhood. It is characterized by unilateral motor impairments that are frequently dominated in the upper limb. In addition to a reduced movement capacity of the affected upper limb, several children with unilateral CP show a reduced awareness of the remaining movement capacity of that limb. This phenomenon of disregarding the preserved capacity of the affected upper limb is regularly referred to as Developmental Disregard (DD). Different theories have been postulated to explain DD, each suggesting slightly different guidelines for therapy. Still, cognitive processes that might additionally contribute to DD in children with unilateral CP have never been directly studied. The current protocol was developed to study cognitive aspects involved in upper limb control in children with unilateral CP with and without DD. This was done by recording event-related potentials (ERPs) extracted from the ongoing EEG during target-response tasks asking for a hand-movement response. ERPs consist of several components, each of them associated with a well-defined cognitive process (e.g., the N1 with early attention processes, the N2 with cognitive control and the P3 with cognitive load and mental effort). Due to its excellent temporal resolution, the ERP technique enables to study several covert cognitive processes preceding overt motor responses and thus allows insight into the cognitive processes that might contribute to the phenomenon of DD. Using this protocol adds a new level of explanation to existing behavioral studies and opens new avenues to the broader implementation of research on cognitive aspects of developmental movement restrictions in children.

Several children with unilateral Cerebral Palsy seem to disregard the preserved capacity of their affected upper limb. This Developmental Disregard is extensively described in the literature but the involved cognitive processes have not been studied. To study underlying cognitive factors of upper limb control, an event-related potential protocol was developed.

Unilateral Cerebral Palsy (CP) is a neurodevelopmental disorder that is a very common cause of disability in childhood. It is characterized by unilateral ...

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer scale. However, current methods mostly rely on tethered cable recordings or only use unidirectional systems, allowing either recording or stimulation of neural activity, but not at the same time or same target. Here, a new wireless, bidirectional device for simultaneous multichannel recording and stimulation of neural activity in freely behaving rats is described. The system operates through a single portable head stage that both transmits recorded activity and can be targeted in real-time for brain stimulation using a telemetry-based multichannel software. The head stage is equipped with a preamplifier and a rechargeable battery, allowing stable long-term recordings or stimulation for up to 1 h. Importantly, the head stage is compact, weighs 12 g (including battery) and thus has minimal impact on the animal&#180;s behavioral repertoire, making the method applicable to a broad set of behavioral tasks. Moreover, the method has the major advantage that the effect of brain stimulation on neural activity and behavior can be measured simultaneously, providing a tool to assess the causal relationships between specific brain activation patterns and behavior. This feature makes the method particularly valuable for the field of deep brain stimulation, allowing precise assessment, monitoring, and adjustment of stimulation parameters during long-term behavioral experiments. The applicability of the system has been validated using the inferior colliculus as a model structure.

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

A wireless, bidirectional system for multi-channel neural recordings and stimulation in freely behaving rats is introduced. The system is light and compact, thus having minimal impact on the animal&#180;s behavioral repertoire. Moreover, this bidirectional system provides a sophisticated tool to assess causal relationships between brain activation patterns and behavior.

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer ...

A Behavioral Assay for Investigating the Role of Spatial Memory During Instinctive Defense in Mice

Evolution has selected a repertoire of defensive behaviors that are essential for survival across all animal species. These behaviors are often stereotyped actions elicited in response to innately aversive sensory stimuli, but their success requires enough flexibility for adapting to different spatial environments, which can change rapidly. Here, we describe a behavioral assay to evaluate the influence of learned spatial knowledge on defensive behaviors in mice. We have adapted the widely used Barnes maze spatial memory assay to investigate how mice navigate to a shelter during escape responses to innately aversive sensory stimuli in a novel environment, and how they adapt to acute changes in the environment. This new assay is an ethological paradigm that does not require training and exploits the natural exploration patterns and navigation strategies in mice. We propose that the set of protocols described here are a powerful means of studying goal-directed behaviors and stimulus-triggered navigation, which should be of interest to both the fields of instinctive behaviors and spatial memory.

This protocol describes a behavioral assay, based on the Barnes maze test, to study how instinctive defensive actions are modified by the knowledge of spatial environment.

Evolution has selected a repertoire of defensive behaviors that are essential for survival across all animal species. These behaviors are often ...

An Automated T-maze Based Apparatus and Protocol for Analyzing Delay- and Effort-based Decision Making in  Free Moving Rodents

Many neurological and psychiatric patients demonstrate difficulties and/or deficits in decision making. Rodent models are helpful to produce a deeper understanding of the neurobiological causes underlying the decision-making problems. A cost-benefit based T-maze task is used for measuring decision making in which rodents choose between a high reward arm (HRA) and a low reward arm (LRA). There are two paradigms of the T-maze decision-making task, one in which the cost is a time delay and the other in which it is physical effort. Both paradigms require a tedious and labor-intensive management of experimental animals, multiple doors, pellet reward, and arm choice recordings. In the current work, we invented an apparatus based on traditional T-maze with full automation for pellet delivery, door management and choice recordings. This automated setup can be used for the evaluation of both delay- and effort-based decision making in rodents. With the protocol described here, our lab investigated the decision-making phenotypes of multiple genetically modified mice. In the representative data, we showed that the mice with ablated medial habenular showed aversions of both delay and effort and tended to choose the immediate and effortless reward. This protocol helps to decrease the variability caused by experimenter intervention and to enhance experiment efficiency. In addition, chronic silicon probe or microelectrode recording, fiber-optic imaging and/or manipulation of neural activity can be easily applied during the decision-making task using the setup described here.

This article introduces an automated T-maze apparatus that we invented, and a protocol based on this apparatus for analyzing delay-based decision making and effort-based decision making in free moving rodents.

Many neurological and psychiatric patients demonstrate difficulties and/or deficits in decision making. Rodent models are helpful to produce a deeper ...

A Visual Guide for Studying Behavioral Defenses to Pathogen Attacks in Leaf-Cutting Ants

The complex lifestyle, evolutionary history of advanced cooperation, and disease defenses of leaf-cutting ants are well studied. Although numerous studies have described the behaviors connected with disease defense, and the associated use of chemicals and antimicrobials, no common visual reference has been made. The main aim of this study was to record short clips of behaviors involved in disease defense, both prophylactically and directly targeted towards an antagonist of the colony following infection. To do so we used an infection experiment, with sub-colonies of the leaf-cutting ant species Acromyrmex echinatior, and the most significant known pathogenic threat to the ants&#39; fungal crop (Leucoagaricus gongylophorus), a specialized pathogenic fungus in the genus Escovopsis. We filmed and compared infected and uninfected colonies, at both early and more advanced stages of infection. We quantified key defensive behaviors across treatments and show that the behavioral response to pathogen attack likely varies between different castes of worker ants, and between early and late detection of a threat. Based on these recordings we have made a library of behavioral clips, accompanied by definitions of the main individual defensive behaviors. We anticipate that such a guide can provide a common frame of reference for other researchers working in this field, to recognize and study these behaviors, and also provide greater scope for comparing different studies to ultimately help better understand the role these behaviors play in disease defense.

We present a visual guide to disease defense behaviors in leaf-cutting ants, with individual clips and accompanying definitions, illustrated in an experimental infection scenario. Our main aim is to help other researchers recognize key defensive behaviors and to provide a common understanding for future research in this field.

The complex lifestyle, evolutionary history of advanced cooperation, and disease defenses of leaf-cutting ants are well studied. Although numerous studies ...