This work was supported by the Humanities Research Institute at Brock University in St. Catharines, Ontario, Canada and by the Cannon Research Center at Carolinas Rehabilitation in Charlotte, North Carolina, USA.

This protocol was approved by the institutional review boards at Brock University and at Carolinas Rehabilitation.
1. Prior to Testing
<ol>
	<li>Create 3 separate lists for the 15 film clips, each listing the clips in a different restricted order.</li>
	<li>Ensure that each list begins and ends with a neutral stimulus and that no two clips that target the same emotion occur consecutively.</li>
	<li>Create a separate folder on the desktop for each list of film order presentation and label the folder with the Order name (e.g., Order 1).</li>
	<li>Save the 15 clips into each of the three folders.</li>
	<li>Re-label each film clip so it reflects the presentation number and gives no clues regarding the target emotion (i.e., 01; 02; 03; etc.).</li>
	<li>Create six randomized orders of task (DANVA-Faces, DANVA-Voices, Film Clips) presentation (see Figure 1).</li>
	<li>Assign the incoming participant to one of the three restricted orders of film clips presentation.</li>
	<li>Assign the incoming participant to one of the six randomized orders of task presentation.</li>
</ol>
2. Day of Testing
<ol>
	<li>Bring participant(s) into the lab and seat them comfortably at the table.</li>
	<li>Review the consent form with the participant by reading each section with him/her.</li>
	<li>Answer any questions the participant may have about the consent form and have him/her sign it.</li>
	<li>Have participants complete the brief demographic questionnaire (Date of birth; gender). Have participants with TBI additionally complete the section of the questionnaire on relevant medical history questionnaire (date of injury, cause of injury, injury severity). 
	NOTE: When participants are completing the questionnaire, inquire about any known visual processing difficulties so you may seat participants with TBI accordingly</li>
	<li>Position the participant in front of the computer in a chair. Ensure that all participants (if more than one) have a clear view of the screen.</li>
</ol>
3. DANVA-Faces Task
<ol>
	<li>Give each participant a clip board with the DANVA-Faces response sheet attached and a pen to circle responses.</li>
	<li>Provide the participant the following instructions: &#34;After you see each item, circle your response (happy, sad, angry, fearful, or neutral) on the line that corresponds to the item number shown on each slide.&#34; 
	NOTE: If participants with TBI have indicated visual processing difficulties or fine motor difficulties are evident, provide the participant with the alternate response page. This page lists the five responses in larger text in landscape format.</li>
	<li>Open the DANVA-Faces task in a presentation and play it using the Slide Show View.</li>
	<li>Give participants the following instructions: &#34;For this activity, I am going to show you some peoples&#39; faces and I want you to tell me how they feel. I want you to tell me if they are happy, sad, angry or fearful. Fearful is the same thing as afraid or scared. If you are unsure of the emotion, choose the &#39;I don&#39;t know&#39; response. There are 24 faces altogether. Each face will be on the screen for five seconds. You must answer before the face disappears for your answer to count. Indicate your answer by circling it on the sheet in front of you&#34;. 
	NOTE: If the participant is unable to circle his/her own responses and is participating in in the study as part of a small group, provide the following instruction: &#34;Indicate your answer by pointing to it on the sheet in front of you&#34;. The examiner will then record the participant&#39;s response on the DANVA-Faces response sheet.</li>
	<li>Ensure the participants do not have any questions before starting.</li>
	<li>Start the task with the two practice trials. Give participants the following instructions: &#34;We are going to complete two practice trials so that you get a sense of how long each face will appear on the screen, and how long you have to provide your answer&#34;.</li>
	<li>Hit enter on the keyboard when the face disappears to move to the next stimulus. Ensure all participants are looking up at the screen before doing so.</li>
	<li>Ensure that the participants do not have questions. If none, begin the test trials. Ensure all participants are looking at the screen. Hit enter to start the test stimuli.</li>
	<li>Continue the procedure outlined for the practice tasks until the 24 stimuli are complete.</li>
	<li>Collect the participant response sheets.</li>
</ol>
4. DANVA-Voices Task
<ol>
	<li>Give each participant a clip board with the DANVA-Voices response sheet attached and a pen to circle responses. 
	NOTE: If participants with TBI have indicated visual processing difficulties or fine motor difficulties are evident, provide the participant with the alternate response page. This page lists the five responses in larger text in landscape format.</li>
	<li>Open the DANVA-Voices task by going to the following website: 
	http://www.psychology.emory.edu/clinical/interpersonal/danva.htm</li>
	<li>Click on the link for Adult faces, voices, and postures.</li>
	<li>Fill in the login and password by entering any letter for the login and typing EMORYDANVA2 for the password.</li>
	<li>Click Continue.</li>
	<li>Click on Voices (green circle in the center of the screen).</li>
	<li>Test the volume level. Provide the following instructions: &#34;I want to be sure that you can hear the sound on the computer and that it is at a comfortable volume for you. I am going to click test sound. Please tell me if you can hear the sound, and also if you need the sound to be louder or more quiet to be comfortable for you.&#34;</li>
	<li>Click test sound.</li>
	<li>Adjust the volume according to the participant&#39;s request (increase/decrease).</li>
	<li>Click &#39;test sound&#39; again. Adjust the volume according to the participant&#39;s request. Do this until the participant reports it is at a comfortable volume.</li>
	<li>Review the task instructions with the participant. Give participants the following instructions: For this activity, you are going to hear someone say the sentence &#39;I&#39;m going out of the room now, but I&#39;ll be back later.&#39; I want you to tell me if they are happy, sad, angry or fearful. Fearful is the same thing as afraid or scared. If you are unsure of the emotion, choose the &#39;I don&#39;t know&#39; response. There are 24 sentences. Before each sentence is spoken, a number will be announced. You need to listen to the sentence that follows. The sentences will not be repeated so you need to listen carefully. Tell me how the person is feeling by circling your answer on the sheet in front of you&#34;. Once you have made your selection, I need to select a response on the computer screen to move to the next item. I will always select the same emotion (fearful). My selection does not indicate the correct answer in anyway.&#34; 
	NOTE: If the participant is unable to circle his/her own responses and is participating in in the study as part of a small group, provide the following instruction: &#34;Indicate your answer by pointing to it on the sheet in front of you&#34;. The examiner will then record the participant&#39;s response on the DANVA-Voices response sheet.</li>
	<li>Click &#39;continue&#39;.</li>
	<li>Direct participants to circle (or point to) their answer on their response sheet when the sentence has played.</li>
	<li>Click &#39;fearful&#39;. Click &#39;next&#39;.</li>
	<li>Continue this procedure until the 24 sentences are complete.</li>
	<li>Exit the website when you reach the end of the task.</li>
	<li>Collect the participants&#39; response sheets.</li>
</ol>
5. Film Clip Task
<ol>
	<li>Seat participants comfortably in front of the computer. Ensure that the screen can be fully seen by all.</li>
	<li>Give each person a clipboard with a copy of the Emotional Film Clip Response sheet. 
	NOTE: If participants with TBI have indicated visual processing difficulties or fine motor difficulties are evident, provide the participant with two alternate response pages. The first page lists the five forced-choice responses and the second provides the 0-9 scale. Both response pages are in larger text and in landscape format.</li>
	<li>Open the folder on the computer for the assigned order of film clip presentation.</li>
	<li>Provide participants with the following directions: &#34;You are going to view a total of 15 film clips. In each film clip, you are going to see a character who looks like they are feeling a certain emotion. They might look &#39;happy&#39;, &#39;sad&#39;, &#39;angry&#39;, &#39;fearful&#39;, or &#39;neutral&#39;. First, I want you to tell me what emotion the character is showing. Then tell me what emotion you experienced while watching the film clip. Finally, tell me the number that best describes the intensity of the emotion you felt while watching the clip. For example, if the person&#39;s face made you feel &#39;mildly&#39; happy or sad, circle a number ranging from 1 to 3. If you felt &#39;moderately&#39; happy or sad, circle a number ranging from 4 to 6. If you felt &#39;very&#39; happy or sad, circle a number ranging from 7 to 9. Only circle 0 if you did not feel any emotion at all. Do you have any questions?&#34; 
	NOTE: Only the first question (Tell me what emotion the character is showing) was analyzed for the current study.</li>
	<li>Double click 01 to open the first film clip.</li>
	<li>Tell the participant which character to focus on while viewing the clip.</li>
	<li>Select &#39;View Full Screen&#39; in the options menu to play the clip.</li>
	<li>Direct the participants to respond to line a (What emotion is the main character experiencing?) on their response sheet when the clip has finished playing.</li>
	<li>Direct the participants to respond to line b (Choose one emotion that best describes how you felt while watching the clip) on the response sheet once line a has been answered.</li>
	<li>Direct the participant to respond to line c (How would you rate the intensity of the emotion you experienced while watching the clip?) once line b has been answered.</li>
	<li>Direct the participant to re-direct his/her attention to the computer screen once the responses are complete.</li>
	<li>Return to the folder on the computer that contains the film clips in the assigned order for the participant.</li>
	<li>Double cck on 02 to open the second film clip. Repeat Steps 5.5 to 5.12 until the participant has viewed all 15 film clips.</li>
	<li>Collect the response sheet from the participant(s).</li>
</ol>
6. Moving from One Task to Another (Optional Step)
<ol>
	<li>Provide participants the option to take a break between Tasks 2 and 3.</li>
</ol>
7. Scoring the Tasks
<ol>
	<li>Refer to the answer key on page 25 of the DANVA2 manual32 to score the DANVA-Faces task.</li>
	<li>Score items that match the answer key as 1 and incorrect items as 0. &#39;I don&#39;t know&#39; responses are scored as 0.</li>
	<li>Calculate a raw score by adding the total number of correct responses. Divide the raw score by 24 (total items) to obtain a percentage accuracy score.</li>
	<li>Refer to the answer key on page 27 of the DANVA2 manual32 and repeat steps 7.2 and 7.3 to score the DANVA-Voices task.</li>
	<li>Refer to the answer key for assigned order of film clip presentation for the participant. Score a 1 if the participant identified the target emotion when responding to the question about what emotion the character was showing, and 0 if the participant did not select the target emotion (including if the participant said &#39;I don&#39;t know&#39;).</li>
	<li>Calculate a raw score by adding the number of correct responses. Divide the raw score by 15 (total items) to obtain a percentage accuracy score.</li>
	<li>Create a subset score for the Film Clip task that contains only responses to the happy, sad, angry, and fearful film clips by adding correct responses to only these 12 clips. Divide the number of correct responses by 12 to obtain a percentage accuracy score.</li>
	<li>Create a score for responses to neutral film clips by adding the number of correct responses to the neutral clips. Divide the number of correct responses by 3 to obtain a percentage accuracy score. 
	NOTE: A subset score needs to be created because the DANVA tasks did not include portrayals of Neutral while the Film Clips task did.</li>
	<li>Calculate a score for responses to positively valenced items (i.e., happy) and negatively valenced items (combine responses to angry, fearful and sad stimuli) for all three tasks.</li>
	<li>Divide the score for positively valenced items for each task by 3 to obtain percentage accuracy score.</li>
	<li>Divide the score for negatively valenced items for each task by 9 to obtain percentage accuracy scores.</li>
</ol>
8. Data Analysis
<ol>
	<li>Conduct a mixed model analyses of variance (ANOVA) to examine responses to isolated facial emotion stimuli, isolated vocal emotion stimuli, and multimodal film clip stimuli by participants with TBI and Control participants.</li>
	<li>Explore main effects found in the ANOVA using follow-up univariate comparisons.</li>
</ol>

The author has nothing to disclose.

The manuscript describes three tasks used to assess emotion recognition abilities of people with TBI. The goal of the described method was to compare response accuracy for facial and vocal emotional cues in isolation, to perception of these cues occurring simultaneously within meaningful situational context. Film clips were included in the current study because their approximation of everyday situations made them more ecologically valid than isolated representations of emotion expressions. When carrying out this protocol...

Traumatic brain injury (TBI) affects approximately 10 million people each year across the world 1. Following TBI, the ability to recognize emotion in others using nonverbal cues such as facial and vocal expressions (i.e., tone of voice), and contextual cues, is often significantly compromised 2-7. Since successful interpersonal interactions and quality relationships are at least partially dependent on accurate interpretation of others&#39; emotions7-10, it is not surprising that difficulty with emotion recognition has been reported to contribute to the poor social outcomes commonly reported following TBI3,8,11,12.
Studies investigating emotion recognition by people with TBI have tended to focus on isolated cues, particularly recognition of facial emotion expressions using static photographs2. While this work has been important in informing the development of treatment programs, it does not adequately represent one&#39;s ability to recognize and interpret nonverbal cues of emotion in everyday interactions. Not only do static images provide increased time to interpret the emotion portrayed, they also typically only portray the apex (i.e., maximum representation) of the expressions13-16. Additionally, static images lack the temporal cues that occur with movement12,13,15, and are not representative of the quickly changing facial expressions of emotion encountered in everyday situations. Further, there is evidence to indicate that static and dynamic visual expressions are processed in different areas of the brain, with more accurate responses to dynamic stimuli17-24.
Vocal emotion recognition by people with TBI has also been studied, both with and without meaningful verbal content. It has been suggested that the presence of verbal content increases perceptual demands because accurate interpretation requires simultaneous processing of the nonverbal tone of voice with the semantic content included in the verbal message25. While many studies have shown improved recognition of vocals affect in the absence of meaningful verbal content, Dimoska et al. 25 found no difference in performance. They argue that people with TBI have a bias toward semantic content in affective sentences; so even when the content is semantically neutral or eliminated, they continue to focus on the verbal information in the sentence. Thus, their results did not clearly show whether meaningful verbal content helps or hinders vocal emotion expression.
In addition to the meaningful verbal content that can accompany the nonverbal vocal cues, context is also provided through the social situation itself. Situational context provides background information about the circumstances under which the emotion expression was produced and can thus greatly influence the interpretation of the expression. When evaluating how someone is feeling, we use context to determine if the situation is consistent with that person&#39;s wants and expectations. This ultimately affects how someone feels, so knowledge of the situational context should result in more accurate inferencing of others&#39; emotion26. The ability to do this is referred to as theory of mind, a construct found to be significantly impaired following TBI6,27-31. Milders et al. 8,31 report that people with TBI have significant difficulty making accurate judgments about and understanding the intentions and feelings of characters in brief stories.
While these studies have highlighted deficits in nonverbal and situational emotion processing, it is difficult to generalize the results to everyday social interactions where these cues occur alongside one another and within a social context. To better understand how people with TBI interpret emotion in everyday social interactions, we need to use stimuli that are multimodal in nature. Only two previous studies have included multimodal stimuli as part of their investigation into how people with TBI process emotion cues. McDonald and Saunders15 and Williams and Wood16 extracted stimuli from the Emotion Evaluation Test (EET), which is part of The&#160;Awareness of Social Inference Test26. The EET consists of videotaped vignettes that show male and female actors portraying nonverbal emotion cues while engaging in conversation. The verbal content in the conversations is semantically neutral; cues regarding how the actors are feeling and contextual information are not provided. Thus, the need to process and integrate meaningful verbal content while simultaneously doing this with the nonverbal facial and vocal cues was eliminated. Results of these studies indicated that people with TBI were significantly less accurate than controls in their ability to identify emotion from multimodal stimuli. Since neither study considered the influence of semantic or contextual information on interpretation of emotion expressions, it remains unclear whether the addition of this type of information would facilitate processing because of increased intersensory redundancy or negatively affect perception because of increased cognitive demands.
This article outlines a set of tasks used to compare perception of facial and vocal emotional cues in isolation, and perception of these cues occurring simultaneously within meaningful situational context. The isolations tasks are part of a larger test of nonverbal emotion recognition &#8212; The Diagnostic Analysis of Nonverbal Accuracy 2 (DANVA2)32. In this protocol, we used the Adult Faces (DANVA-Faces) and Adult Paralanguage (DANVA-Voices) subtests. Each of these subtests includes 24 stimuli, depicting six representations each of happy, sad, angry, and fearful emotions. The DANVA2 requires participants to identify the emotion portrayed using a forced-choice format (4 choices). Here, a fifth option (I don&#39;t know) is provided for both subtests. The amount of time each facial expression was shown was increased from two seconds to five seconds to ensure that we were assessing affect recognition and not speed of processing. We did not alter the adult paralanguage subtest beyond adding the additional choice in the forced-choice response format. Participants heard each sentence one time only. Feedback was not provided in either task.
Film clips were used to assess perception of facial and vocal emotion cues occurring simultaneously within situational context. While the verbal content within these clips did not explicitly state what the characters in the clips were feeling, it was meaningful to the situational context. Fifteen clips extracted from commercial movies and television series were included in the study, each ranging from 45 to 103 (mean = 71.87) seconds. Feedback was not provided to participants during this task. There were three clips representing each of the following emotion categories: happy, sad, angry, fearful, neutral. These clips were chosen based on results of a study conducted with young adults (n = 70)&#160;Zupan, B. &#38; Babbage, D. R. Emotion elicitation stimuli from film clips and narrative text. Manuscript submitted to J Soc Psyc. In that study, participants were presented with six film clips for each of the target emotion categories, and six neutral film clips, ranging from 24 to 127 seconds (mean = 77.3 seconds). While the goal was to include relatively short film clips, each clip chosen for inclusion needed to have sufficient contextual information that it could be understood by viewers without having additional knowledge of the film from which it was taken33.
The clips selected for the current study had been correctly identified in the Zupan and Babbage study Emotion elicitation stimuli from film clips and narrative text, cited above as the target emotion between 83 and 89% for happy, 86 - 93% for sad, 63 - 93% for angry, 77 - 96% for fearful, and 81 - 87% for neutral. While having the same number of exemplars of each emotion category as was in the DANVA2 tasks (n = 6) would have been ideal, we opted for only three due to the increased length of the film clip stimuli compared to stimuli in the DANVA-Faces and DANVA-Voices tasks.
Data from these tasks were collected from 60 participants with TBI and 60 age and gender matched controls. Participants were seen either individually or in small groups (max = 3) and the order of the three tasks was randomized across testing sessions.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Diagnostic Analysis of Nonverbal Accuracy-2</td>
			<td>Department of Pychology, Emory University. Atlanta, GA</td>
			<td></td>
			<td>DANVA-Faces subtest, DANVA-Voices subtest</td>
		</tr>
		<tr>
			<td>Computer</td>
			<td>Apple iMac Desktop, 27&#34; display</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Statistical Analysis Software</td>
			<td>SPSS</td>
			<td></td>
			<td>University Licensed software for data analysis</td>
		</tr>
		<tr>
			<td>Happy Film Clip 1</td>
			<td>Sweet Home Alabama, D&#38;D Films, 2002, Director: A Tennant</td>
			<td></td>
			<td>A man surprises his girlfriend by proposing in a jewelry store</td>
		</tr>
		<tr>
			<td>Happy Film Clip 2</td>
			<td>Wedding Crashers, New Line Cinema, 2005, Director: D. Dobkin</td>
			<td></td>
			<td>A couple is sitting on the beach and flirting while playing a hand game</td>
		</tr>
		<tr>
			<td>Happy Film Clip 3</td>
			<td>Reme-mber the Titans, Jerry Bruckheimer Films, 2000, Director: B. Yakin</td>
			<td></td>
			<td>An African American football coach and his family are accepted by their community when the school team wins the football championship</td>
		</tr>
		<tr>
			<td>Sad Film Clip 1</td>
			<td>Grey&#39;s Anatomy, ABC, 2006, Director: P. Horton</td>
			<td></td>
			<td>A father is only able to communicate with his family using his eyes.&#160;</td>
		</tr>
		<tr>
			<td>Sad Film Clip 2</td>
			<td>Armageddon, Touchstone Pictures, 1998, Director: M. Bay</td>
			<td></td>
			<td>A daughter is saying goodbye to her father who is in space on a dangerous mission</td>
		</tr>
		<tr>
			<td>Sad Film Clip 3</td>
			<td>Grey&#39;s Anatomy, ABC, 2006, Director: M. Tinker</td>
			<td></td>
			<td>A woman is heartbroken her fiance has died</td>
		</tr>
		<tr>
			<td>Angry Film Clip 1</td>
			<td>Anne of Green Gables, Canadian Broadcast Corporation, 1985, Director: K. Sullivan</td>
			<td></td>
			<td>An older woman speaks openly about a child&#39;s physical appearance in front of her</td>
		</tr>
		<tr>
			<td>Angry Film Clip 2</td>
			<td>Enough, Columbia Pictures, 2002, Director: M. Apted</td>
			<td></td>
			<td>A wife confronts her husband about an affair when she smells another woman&#39;s perfume on his clothing</td>
		</tr>
		<tr>
			<td>Angry Film Clip 3</td>
			<td>Pretty Woman, Touchstone Pictures, 1990, Director: G. Marshall</td>
			<td></td>
			<td>A call girl attempts to purchase clothing in an expensive boutique and is turned away</td>
		</tr>
		<tr>
			<td>Fearful Film Clip 1</td>
			<td>Blood Diamond, Warner Bros Pictures, 2006, Director: E. Zwick</td>
			<td></td>
			<td>Numerous vehicles carrying militia approach a man and his son while they are out walking</td>
		</tr>
		<tr>
			<td>Fearful Film Clip 2</td>
			<td>The Life Before Her Eyes, 2929 Productions, 2007, Directors: V. Perelman</td>
			<td></td>
			<td>Two teenaged gilrs are talking in the school bathroom when they hear gunshots that continue to get closer</td>
		</tr>
		<tr>
			<td>Fearful Film Clip 3</td>
			<td>The Sixth Sense, Barry Mendel Productions, 1999, Director: M. N. Shyamalan</td>
			<td></td>
			<td>A young boy enters the kitchen in the middle of the night and finds his mother behaving very atypically</td>
		</tr>
		<tr>
			<td>Neutral Film Clip 1</td>
			<td>The Other Sister, Mandeville Films, 1999, Director: G. Marshall</td>
			<td></td>
			<td>A mother and daughter are discussing an art book while sitting in their living room</td>
		</tr>
		<tr>
			<td>Neutral Film Clip 2</td>
			<td>The Game, Polygram Filmed Entertainment, 1999, Director: D. Fincher</td>
			<td></td>
			<td>One gentleman is explaining the rules of a game to another</td>
		</tr>
		<tr>
			<td>Neutral Film Clip 3</td>
			<td>The Notebook, New Line Cinema, 2004, Director: N. Cassavetes</td>
			<td></td>
			<td>An older gentleman and his doctor are conversing during a routine check-up</td>
		</tr>
	</tbody>
</table>

exploring the use of isolated expressions and film clips to evaluate emotion recognition by people with traumatic brain injury

The current study presented 60 people with traumatic brain injury (TBI) and 60 controls with isolated facial emotion expressions, isolated vocal emotion expressions, and multimodal (i.e., film clips) stimuli that included contextual cues. All stimuli were presented via computer. Participants were required to indicate how the person in each stimulus was feeling using a forced-choice format. Additionally, for the film clips, participants had to indicate how they felt in response to the stimulus, and the level of intensity with which they experienced that emotion.

This task battery was used to compare emotion recognition for isolated emotion expressions (i.e., face-only; voice-only) and combined emotion expressions (i.e., face and voice) that occur within a situational context. A total of 60 (37 males and 23 females) participants with moderate to severe TBI between the ages of 21 and 63 years (mean = 40.98) and 60 (38 males and 22 females) age -matched Controls (range = 18 to 63; mean = 40.64) completed the three tasks. To partici...

Watch this Scientific Journal Video about Exploring the Use of Isolated Expressions and Film Clips to Evaluate Emotion Recognition by People with Traumatic Brain Injury at JoVE.com

Exploring the Use of Isolated Expressions and Film Clips to Evaluate Emotion Recognition by People with Traumatic Brain Injury

This paper describes how to implement a battery of behavioral tasks to examine emotion recognition of isolated facial and vocal emotion expressions, and a novel task using commercial television and film clips to assess multimodal emotion recognition that includes contextual cues.

The current study presented 60 people with traumatic brain injury (TBI) and 60 controls with isolated facial emotion expressions, isolated vocal emotion ...

exploring-use-isolated-expressions-film-clips-to-evaluate-emotion

Research

JoVE Journal

Behavior

8.9K Views.  Brock University. This paper describes how to implement a battery of behavioral tasks to examine emotion recognition of isolated facial and vocal emotion expressions, and a novel task using commercial television and film clips to assess multimodal emotion recognition that includes contextual cues.

Video: Exploring the Use of Isolated Expressions and Film Clips to Evaluate Emotion Recognition by People with Traumatic Brain Injury

Use of the Operant Orofacial Pain Assessment Device (OPAD) to Measure Changes in Nociceptive Behavior

We present an operant system for the detection of pain in awake, conscious rodents. The Orofacial Pain Assessment Device (OPAD) assesses pain behaviors in a more clinically relevant way by not relying on reflex-based measures of nociception. Food fasted, hairless (or shaved) rodents are placed into a Plexiglas chamber which has two Peltier-based thermodes that can be programmed to any temperature between 7 &deg;C and 60 &deg;C. The rodent is trained to make contact with these in order to access a reward bottle. During a session, a number of behavioral pain outcomes are automatically recorded and saved. These measures include the number of reward bottle activations (licks) and facial contact stimuli (face contacts), but custom measures like the lick/face ratio (total number of licks per session/total number of contacts) can also be created. The stimulus temperature can be set to a single temperature or multiple temperatures within a session. The OPAD is a high-throughput, easy to use operant assay which will lead to better translation of pain research in the future as it includes cortical input instead of relying on spinal reflex-based nociceptive assays.

Use of the Operant Orofacial Pain Assessment Device OPAD to Measure Changes in Nociceptive Behavior

We present a user-friendly, high-throughput operant system for the evaluation of pain behaviors in awake, conscious rodents. The Orofacial Pain Assessment Device (OPAD) can assess pain through a reward/conflict paradigm thus providing a more humane way of testing. This protocol will yield more clinically relevant and translational data from rodents.

We present an operant  system for the detection of pain in awake, conscious rodents. The Orofacial  Pain Assessment Device (OPAD) assesses pain behaviors ...

Use of an Eight-arm Radial Water Maze to Assess Working and Reference Memory Following Neonatal Brain Injury

Working and reference memory are commonly assessed using the land based radial arm maze.&#160;However, this paradigm requires pretraining, food deprivation, and may introduce scent cue confounds. The eight-arm radial water maze is designed to evaluate reference and working memory performance simultaneously by requiring subjects to use extra-maze cues to locate escape platforms and remedies the limitations observed in land based radial arm maze designs. Specifically, subjects are required to avoid the arms previously used for escape during each testing day (working memory) as well as avoid the fixed arms, which never contain escape platforms (reference memory). Re-entries into arms that have already been used for escape during a testing session (and thus the escape platform has been removed) and re-entries into reference memory arms are indicative of working memory deficits. Alternatively, first entries into reference memory arms are indicative of reference memory deficits. We used this maze to compare performance of rats with neonatal brain injury and sham controls following induction of hypoxia-ischemia and show significant deficits in both working and reference memory after eleven days of testing. This protocol could be easily modified to examine many other models of learning impairment.

The eight-arm radial water maze is designed to evaluate reference and working memory performance simultaneously by requiring subjects to use extra-maze cues to locate escape platforms and remedies the limitations observed in land based radial arm maze designs.

Working and reference memory are commonly assessed using the land based radial arm maze.&#160;However, this paradigm requires pretraining, food ...

Uncovering Beat Deafness: Detecting Rhythm Disorders with Synchronized Finger Tapping and Perceptual Timing Tasks

A set of behavioral tasks for assessing perceptual and sensorimotor timing abilities in the general population (i.e., non-musicians) is presented here with the goal of uncovering rhythm disorders, such as beat deafness. Beat deafness is characterized by poor performance in perceiving durations in auditory rhythmic patterns or poor synchronization of movement with auditory rhythms (e.g., with musical beats). These tasks include the synchronization of finger tapping to the beat of simple and complex auditory stimuli and the detection of rhythmic irregularities (anisochrony detection task) embedded in the same stimuli. These tests, which are easy to administer, include an assessment of both perceptual and sensorimotor timing abilities under different conditions (e.g., beat rates and types of auditory material) and are based on the same auditory stimuli, ranging from a simple metronome to a complex musical excerpt. The analysis of synchronized tapping data is performed with circular statistics, which provide reliable measures of synchronization accuracy (e.g., the difference between the timing of the taps and the timing of the pacing stimuli) and consistency. Circular statistics on tapping data are particularly well-suited for detecting individual differences in the general population. Synchronized tapping and anisochrony detection are sensitive measures for identifying profiles of rhythm disorders and have been used with success to uncover cases of poor synchronization with spared perceptual timing. This systematic assessment of perceptual and sensorimotor timing can be extended to populations of patients with brain damage, neurodegenerative diseases (e.g., Parkinson&#8217;s disease), and developmental disorders (e.g., Attention Deficit Hyperactivity Disorder).

Behavioral tasks that allow for the assessment of perceptual and sensorimotor timing abilities in the general population (i.e., non-musicians) are presented. Synchronization of finger tapping to the beat of an auditory stimuli and detecting rhythmic irregularities provides a means of uncovering rhythm disorders.

A set of behavioral tasks for assessing perceptual and sensorimotor timing abilities in the general population (i.e., non-musicians) is presented here ...

Assessing Spatial Memory Impairment in a Mouse Model of Traumatic Brain Injury Using a Radial Water Tread Maze

Despite the recent increase in use of mouse models in scientific research, researchers continue to use cognitive tasks that were originally designed and validated for rat use. The Radial Water Tread (RWT) maze test of spatial memory (designed specifically for mice and requiring no swimming) has been shown previously to successfully distinguish between controlled cortical impact-induced TBI mice and sham controls. Here, a detailed protocol for this task is presented. The RWT maze capitalizes on the natural tendency of mice to avoid open areas in favor of hugging the sides of an apparatus (thigmotaxis). The walls of the maze are lined with nine escape holes placed above the floor of the apparatus, and mice are trained to use visual cues to locate the escape hole that leads out of the maze. The maze is filled with an inch of cold water, sufficient to motivate escape but not deep enough to require that the mouse swim. The acquisition period takes only four training days, with a test of memory retention on day five and a long-term memory test on day 12. The results reported here suggest that the RWT maze is a feasible alternative to rat-validated, swimming-based cognitive tests in the assessment of spatial memory deficits in mouse models of TBI.

Here we present a protocol for a mouse-specific test of cognition that does not require swimming. This test can be used to successfully distinguish controlled cortical impact-induced traumatic brain injury mice from sham controls.

Despite the recent increase in use of mouse models in scientific research, researchers continue to use cognitive tasks that were originally designed and ...

Detecting Behavioral Deficits in Rats After Traumatic Brain Injury

With the increasing incidence of traumatic brain injury (TBI) in both civilian and military populations, TBI is now considered a chronic disease; however, few studies have investigated the long-term effects of injury in rodent models of TBI. Shown here are behavioral measures that are well-established in TBI research for times early after injury, such as two weeks, until two months. Some of these methods have previously been used at later times after injury, up to one year, but by very few laboratories. The methods demonstrated here are a short neurological assessment to test reflexes, a Beam-Balance to test balance, a Beam-Walk to test balance and motor coordination, and a working memory version of the Morris water maze that can be sensitive to deficits in reference memory. Male rats were handled and pre-trained to neurological, balance, and motor coordination tests prior to receiving parasagittal fluid percussion injury (FPI) or sham injury. Rats can be tested on the short neurological assessment (neuroscore), the beam-balance, and the Beam-Walk multiple times, while testing on the water maze can only be done once. This difference is because rats can remember the task, thus confounding the results if repeated testing is attempted in the same animal. When testing from one to three days after injury, significant differences are detected in all three non-cognitive tasks. However, differences in the Beam-Walk task were not detectable at later time points (after 3 months). Deficits were detected at 3 months in the Beam-Balance and at 6 months in the neuroscore. Deficits in working memory were detected out to 12 months after injury, and a deficit in a reference memory first appeared at 12 months. Thus, standard behavioral tests can be useful measures of persistent behavioral deficits after FPI.

The goal of the behavioral tests presented here is to detect functional deficits in rats after traumatic brain injury. Four specific tests are presented that detect deficits in behaviors to reflect the damage to specific brain areas at times extending to one year after injury.

With the increasing incidence of traumatic brain injury (TBI) in both civilian and military populations, TBI is now considered a chronic disease; however, ...

Using Laser Doppler Imaging and Monitoring to Analyze Spinal Cord Microcirculation in Rat

Laser Doppler flowmetry (LDF) is a noninvasive method for blood flow (BF) measurement, which makes it preferable for measuring microcirculatory alterations of the spinal cord. In this article, our goal was to use both Laser Doppler imaging and monitoring to analyze the change of BF after spinal cord injury. Both the laser Doppler image scanner and the probe/monitor were being employed to obtain each readout. The data of LDPI provided a local distribution of BF, which gave an overview of perfusion around the injury site and made it accessible for comparative analysis of BF among different locations. By intensely measuring the probing area over a period of time, a combined probe was used to simultaneously measure the BF and oxygen saturation of the spinal cord, showing overall spinal cord perfusion and oxygen supply. LDF itself has a few limitations, such as relative flux, sensitivity to movement, and biological zero signal. However, the technology has been applied in clinical and experimental study due to its simple setup and rapid measurement of BF.

Here we present a combination of laser Doppler perfusion imaging (LDPI) and laser Doppler perfusion monitoring (LDPM) to measure spinal cord local blood flows and oxygen saturation (SO2), as well as a standardized procedure for introducing spinal cord trauma on rat.

Laser Doppler flowmetry (LDF) is a noninvasive method for blood flow (BF) measurement, which makes it preferable for measuring microcirculatory ...

Novel Object Recognition and Object Location Behavioral Testing in Mice on a Budget

Ethologically relevant behavioral testing is a critical component of any study that uses mouse models to study the cognitive effects of various physiological or pathological changes. The object location task (OLT) and the novel object recognition task (NORT) are two effective behavioral tasks commonly used to reveal the function and relative health of specific brain regions involved in memory. While both of these tests exploit the inherent preference of mice for the novelty to reveal memory for previously encountered objects, the OLT primarily evaluates spatial learning, which relies heavily on hippocampal activity. The NORT, in contrast, evaluates non-spatial learning of object identity, which relies on multiple brain regions. Both tasks require an open-field-testing arena, objects with equivalent intrinsic value to mice, appropriate environmental cues, and video recording equipment and the software. Commercially available systems, while convenient, can be costly. This manuscript details a simple, cost-effective method for building the arenas and setting up the equipment necessary to perform the OLT and NORT. Furthermore, the manuscript describes an efficient testing protocol that incorporates both OLT and NORT and provides typical methods for data acquisition and analysis, as well as representative results. Successful completion of these tests can provide valuable insight into the memory function of various mouse model systems and appraise the underlying neural regions that support these functions.

Here we provide a protocol which includes comprehensive instructions for the economical establishment of murine object location and novel object recognition behavioral testing, including the design, cost, and construction of required equipment as well as execution of behavioral testing, data collection, and analysis.

Ethologically relevant behavioral testing is a critical component of any study that uses mouse models to study the cognitive effects of various ...

Exploring Infant Sensitivity to Visual Language using Eye Tracking and the Preferential Looking Paradigm

We discuss the use of the preferential looking paradigm in eye tracking studies in order to study how infants develop, understand, and attend to the world around them. Eye tracking is a safe and non-invasive way to collect gaze data from infants, and the preferential looking paradigm is simple to design and only requires the infant to be attending to the screen. By simultaneously showing two visual stimuli that differ in one dimension, we can assess whether infants show different looking behavior for either stimulus, thus demonstrating sensitivity to that difference. The challenges in such experimental approaches are that experiments must be kept brief (no more than 10 min) and be carefully controlled such that the two stimuli differ in only one way. The interpretation of null results must also be carefully considered. In this paper, we illustrate a successful example of an infant eye tracking study with a preferential looking paradigm to discover that 6-month-olds are sensitive to linguistic cues in a signed language despite having no prior exposure to signed language, suggesting that infants possess intrinsic or innate sensitivities to these cues.

Eye tracking studies using a preferential looking paradigm can be used to study infants&#39;&#160;emerging understanding of, and attention to, their external visual world.

We discuss the use of the preferential looking paradigm in eye tracking studies in order to study how infants develop, understand, and attend to the world ...

Use of a Wireless Video-EEG System to Monitor Epileptiform Discharges Following Lateral Fluid-Percussion Induced Traumatic Brain Injury

The lateral fluid percussion injury (FPI) model is well established and has been used to study TBI and post-traumatic epilepsy (PTE). However, considerable variability has been reported for the specific parameters used in different studies that have employed this model, making it difficult to harmonize and interpret the results between laboratories. For example, variability has been reported regarding the size and location of the craniectomy, how the Luer lock hub is placed relative to the craniectomy, the atmospheric pressure applied to the dura and the duration of the pressure pulse. Each of these parameters can impact injury severity, which directly correlates with the incidence of PTE. This has been manifested as a wide range of mortality rates, righting reflex times and incidence of convulsive seizures reported. Here we provide a detailed protocol for the method we have used to help facilitate harmonization between studies. We used FPI in combination with a wireless EEG telemetry system to continuously monitor for electrographic changes and detect seizure activity.&#160; FPI is induced by creating a 5 mm craniectomy over the left hemisphere, between the Bregma and Lambda and adjacent to the lateral ridge. A Luer lock hub is secured onto the skull over the craniectomy. This hub is connected to the FPI device, and a 20-millisecond pressure pulse is delivered directly to the intact dura through pressure tubing connected to the hub via a twist lock connector. Following recovery, rats are re-anesthetized to remove the hub. Five 0.5 mm, stainless steel EEG electrode screws are placed in contact with the dura through the skull and serve as four recording electrodes and one reference electrode. The electrode wires are collected into a pedestal connector which is secured into place with bone cement. Continuous video/EEG recordings are collected for up to 4 weeks post TBI.

Here we present a protocol to induce severe TBI with the lateral fluid percussion injury (FPI) model in adult, male Wistar rats. We also demonstrate the use of a wireless telemetry system to collect continuous video-EEG recordings and monitor for epileptiform discharges consistent with post-traumatic epileptogenesis.&#160;

The lateral fluid percussion injury (FPI) model is well established and has been used to study TBI and post-traumatic epilepsy (PTE). However, ...

Inducing Post-Traumatic Epilepsy in a Mouse Model of Repetitive Diffuse Traumatic Brain Injury

Traumatic brain injury (TBI) is a leading cause of acquired epilepsy. TBI can result in a focal or diffuse brain injury. Focal injury is a result of direct mechanical forces, sometimes penetrating through the cranium, creating a direct lesion in the brain tissue. These are visible during brain imaging as areas with contusion, laceration, and hemorrhage. Focal lesions induce neuronal death and glial scar formation and are present in 20%&minus;25% of all people who incur a TBI. However, in the majority of TBI cases, injury is caused by acceleration-deceleration forces and subsequent tissue shearing, resulting in nonfocal, diffuse damage. A subpopulation of TBI patients continues to develop post-traumatic epilepsy (PTE) after a latency period of months or years. Currently, it is impossible to predict which patients will develop PTE, and seizures in PTE patients are challenging to control, necessitating further research. Until recently, the field was limited to only two animal/rodent models with validated spontaneous post-traumatic seizures, both presenting with large focal lesions with massive tissue loss in the cortex and sometimes subcortical structures. In contrast to these approaches, it was determined that diffuse TBI induced using a modified weight drop model is sufficient to initiate development of spontaneous convulsive and non-convulsive seizures, even in the absence of focal lesions or tissue loss. Similar to human patients with acquired post-traumatic epilepsy, this model presents with a latency period after injury before seizure onset. In this protocol, the community will be provided with a new model of post-traumatic epilepsy, detailing how to induce diffuse non-lesional TBI followed by continuous long-term video-electroencephalographic animal monitoring over the course of several months. This protocol will detail animal handling, the weight drop procedure, the electrode placement for two acquisition systems, and the frequent challenges encountered during each of the steps of surgery, postoperative monitoring, and data acquisition.

This systematic protocol describes a new animal model of post-traumatic epilepsy after repetitive mild traumatic brain injury. The first part details steps for traumatic brain injury induction using a modified weight drop model. The second part provides instructions on the surgical approach for single- and multi-channel electroencephalographic data acquisition systems.

Traumatic brain injury (TBI) is a leading cause of acquired epilepsy. TBI can result in a focal or diffuse brain injury. Focal injury is a result of ...