Name: neuroimaging field methods using functional near infrared spectroscopy (nirs) neuroimaging to study global child development: rural sub-saharan africa
Uploaded: 2018-02-02T15:00:00
Description: Watch this Scientific Journal Video about Neuroimaging Field Methods Using Functional Near Infrared Spectroscopy NIRS Neuroimaging to Study Global Child Development: Rural Sub-Saharan Africa at JoVE.c

This research was made possible through the Jacobs Foundation Early Career Fellowship to K. Jasinska (Fellowship Number: 2015 118455). The authors also wish to acknowledge Axel Blahoua, Fabrice Tanoh, Ariane Amon, Brice Kanga, and Yvette Foto for their assistance in data collection and field support. Special thanks to the families and children of Moap&#233;, Anangui&#233;, Affery, and Becouefin for their participation in this research program and the villages&#39; warm hospitality.

All methods described here have been approved by the Institutional Review Board (IRB) of the University of Delaware.
1. Mobile Laboratory Transport and Setup
<ol>
	<li>Traveling with the fNIRS equipment
	<ol>
		<li>Transport fNIRS equipment. 
		NOTE: fNIRS equipment can be transported as checked-luggage on a major international airline, but it is imperative to confirm with the given airline. Equipment restrictions may vary by origin or destination country. Alternatively...

<ol>
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E., Jung, J., Jang, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NIRS-SPM:+statistical+parametric+mapping+for+near-infrared+spectroscopy.">NIRS-SPM: statistical parametric mapping for near-infrared spectroscopy.</a> Neuroimage. 44 (2), 428-447 (2009).</li><li>Huppert, T. J. T. J., Diamond, S. G. S. G., Franceschini, M. A. M. A., Boas, D. A. D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HomER:+a+review+of+time-series+analysis+methods+for+near-infrared+spectroscopy+of+the+brain.">HomER: a review of time-series analysis methods for near-infrared spectroscopy of the brain.</a> Appl Opt. 48 (10), D280-D298 (2009).</li><li>Huppert, T. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Commentary+on+the+statistical+properties+of+noise+and+its+implication+on+general+linear+models+in+functional+near-infrared+spectroscopy.">Commentary on the statistical properties of noise and its implication on general linear models in functional near-infrared spectroscopy.</a> Neurophotonics. 3 (1), 010401 (2016).</li><li>Rosso, A. 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=Neuroimaging+of+an+attention+demanding+dual-task+during+dynamic+postural+control.">Neuroimaging of an attention demanding dual-task during dynamic postural control.</a> Gait Posture. 57, 193-198 (2017).</li><li>Jang, K. E. K. E., 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=Wavelet+minimum+description+length+detrending+for+near-infrared+spectroscopy.">Wavelet minimum description length detrending for near-infrared spectroscopy.</a> Journal of Biomedical Optics. 14 (3), 034004-034004 (2009).</li><li>Worsley, K. J., Friston, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+fMRI+time-series+revisited--again.">Analysis of fMRI time-series revisited--again.</a> Neuroimage. 2 (3), 173-181 (1995).</li><li>Friston, K. 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K., Okamoto, M., Dan, H., Jurcak, V., Dan, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatial+registration+of+multichannel+multi-subject+fNIRS+data+to+MNI+space+without+MRI.">Spatial registration of multichannel multi-subject fNIRS data to MNI space without MRI.</a> Neuroimage. 27 (4), 842-851 (2005).</li><li>Krosin, M. T., Klitzman, R., Levin, B., Cheng, J., Ranney, M. 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This paper presented a field neuroimaging protocol suitable for low-resource contexts in remote locations. The key advance of this field neuroimaging protocol is the first-time ability to study brain function and its development in understudied (or never-before studied) contexts. Critical steps in this protocol include traveling with and setting up a mobile laboratory suitable for quality data collection in tropical climates without electricity or available facilities. This protocol provides a general guide to forming st...

Portable fNIRS imaging provides the ability to study brain function and development outside the laboratory, in previously inaccessible settings or with understudied populations. Much of the knowledge in the domain of cognitive neuroscience comes from imaging studies conducted in university or hospital laboratory settings, in predominantly Western countries. By design, this contributes to a seldom-spoken-of problem in research: much of what is known about the brain is based on studies with participants for whom laboratory settings in (mostly) Western countries are accessible. That is, most neuroimaging research involves participants who live in reasonable proximity to a neuroimaging laboratory and have both the time and resources necessary to participate in a study. As a discipline, cognitive neuroscience aims to understand the brain and the factors that shape its development&#8212;including the powerful effects of a child&#39;s environment and their early-life experiences1,2,3. Methods that advance the field&#39;s capacity to study development in a fuller range of human experience can dramatically advance the understanding of the complex relation between brain development and the life experiences that shape it.
This paper presents a protocol for field neuroimaging, which was developed for use in rural sub-Saharan Africa, specifically southern C&#244;te d&#39;Ivoire. The aim of this field neuroimaging research program was to understand children&#39;s reading development in an environment with a high-risk of illiteracy. C&#244;te d&#39;Ivoire&#39;s youth (15-24 years) literacy rate is 53%, despite 93% primary school enrollment rates4. C&#244;te d&#39;Ivoire is the world&#39;s primary source of cocoa, and there are an estimated 1.3 million child laborers in the cocoa agricultural sector5. Yet, little is known about the impact of child labor on brain development and learning, specifically learning to read. Applying the latest tools of cognitive neuroscience, i.e., portable neuroimaging methods,&#160;can yield valuable insights into children&#8217;s learning outcomes.&#160;For example, field neuroimaging with fNIRS can allow the identification of neurodevelopmental periods during which targeted educational programs or interventions may have maximal impacts on children&#39;s learning outcomes.
fNIRS neuroimaging is well-suited for field research. Similar to functional magnetic resonance imaging (fMRI), fNIRS measures the brain&#39;s hemodynamic response6. However, fNIRS uses a series of light emitting optodes and light detectors rather than generating electromagnetic fields. There are no restrictions on metal in or near the testing area, and no electric shielding is necessary, as in the case for electroencephalography (EEG). A key advantage of fNIRS is its portability (i.e., some systems may fit in a suitcase) and ease of use. fNIRS is also easy to use with children; the child is comfortably seated in a chair during the experiment and the fNIRS system tolerates movement well compared to fMRI. Compared with fMRI, fNIRS also provides separate measures of deoxygenated (HbR) and oxygenated hemoglobin (HbO) during recording, compared to fMRI which yields a combined blood oxygen level density (BOLD) measure. fNIRS has superior temporal resolution to fMRI: sampling rates can vary between ~ 7-15 Hz. fNIRS has good spatial resolution: the fNIRS&#39; depth of recording in the human cortex is less than fMRI, measuring about 3 to 4 cm in depth, which is well-suited for studying cortical functions, especially with infants and children who have thinner skulls than adults3,7,8,9,10.
This field neuroimaging protocol outlines considerations for traveling with and setting up a portable neuroimaging laboratory in low-resource contexts. The protocol also highlights the essential nature of meaningful, long-term collaborations with local science partners and ways by which this approach serves to build local science capacity. The neuroimaging protocol for collecting and analyzing fNIRS brain data from a battery of language, reading, and cognitive tasks, is demonstrated including recommendations for creating culturally appropriate informed consent procedures for imaging research. While this protocol is designed for cognitive development research with primary school aged children in rural C&#244;te d&#39;Ivoire, the protocol is highly relevant for any field neuroimaging study in challenging, low-resource environments, and can be adapted for novel contexts.

<table>
	<tbody>
		<tr>
			<td>LIGHTNIRS Main Unit Pack 120V</td>
			<td>Shimadzu</td>
			<td>292-34000-42</td>
			<td>Component of the fNIRS system</td>
		</tr>
		<tr>
			<td>HOLDER ASSY, ALL- CAP</td>
			<td>Shimadzu</td>
			<td>594-07618-01</td>
			<td>Component of the fNIRS system</td>
		</tr>
		<tr>
			<td>LIGHTNIRS connection cable</td>
			<td>Shimadzu</td>
			<td>567-10976-11</td>
			<td>fNIRS system component</td>
		</tr>
		<tr>
			<td>Fiber set for LIGHTNIRS, 1m (8 sets)</td>
			<td>Shimadzu</td>
			<td>567-11350-01</td>
			<td>fNIRS system component</td>
		</tr>
		<tr>
			<td>Dell Latitude Laptop</td>
			<td>Shimadzu (from Dell)</td>
			<td>220-97322-00</td>
			<td>Master computer to run fNIRS applications</td>
		</tr>
		<tr>
			<td>PATRIOT SEU (System Electronics Unit)</td>
			<td>POLHEMUS</td>
			<td>1A0453-001</td>
			<td>PATRIOT System component</td>
		</tr>
		<tr>
			<td>Power Supply</td>
			<td>POLHEMUS</td>
			<td>2C0809</td>
			<td>PATRIOT System component</td>
		</tr>
		<tr>
			<td>Power Supply cord</td>
			<td>POLHEMUS</td>
			<td>17500B-BLK</td>
			<td>PATRIOT System component</td>
		</tr>
		<tr>
			<td>RS-232 null modem cable</td>
			<td>POLHEMUS</td>
			<td>1C0288</td>
			<td>PATRIOT System component</td>
		</tr>
		<tr>
			<td>USB cable</td>
			<td>POLHEMUS</td>
			<td>1C0289</td>
			<td>PATRIOT System component</td>
		</tr>
		<tr>
			<td>RX2 Sensor 10&#39; cable</td>
			<td>POLHEMUS</td>
			<td>4A0492-20</td>
			<td>PATRIOT System component</td>
		</tr>
		<tr>
			<td>TX2 Source 10&#39; cable</td>
			<td>POLHEMUS</td>
			<td>4A0506-20</td>
			<td>PATRIOT System component</td>
		</tr>
	</tbody>
</table>

neuroimaging field methods using functional near infrared spectroscopy (nirs) neuroimaging to study global child development: rural sub-saharan africa

Portable neuroimaging approaches provide new advances to the study of brain function and brain development with previously inaccessible populations and in remote locations. This paper shows the development of field functional Near Infrared Spectroscopy (fNIRS) imaging to the study of child language, reading, and cognitive development in a rural village setting of C&#244;te d&#39;Ivoire. Innovation in methods and the development of culturally appropriate neuroimaging protocols allow&#160;a first-time look into the brain&#39;s development and children&#39;s learning outcomes in understudied environments. This paper demonstrates protocols for transporting and setting up a mobile laboratory, discusses considerations for field versus laboratory neuroimaging, and presents a guide for developing neuroimaging consent procedures and building meaningful long-term collaborations with local government and science partners. Portable neuroimaging methods can be used to study complex child development contexts, including the impact of significant poverty and adversity on brain development. The protocol presented here has been developed for use in C&#244;te d&#39;Ivoire, the world&#39;s primary source of cocoa, and where reports of child labor in the cocoa sector are common. Yet, little is known about the impact of child labor on brain development and learning. Field neuroimaging methods have the potential to yield new insights into such urgent issues, and the development of children globally.

Probe position data obtained by the 3D digitizer (Figure 2) can be visualized on a standard brain template. Register fNIRS channels to MNI space using NIRS-SPM&#39;s stand-alone registration function25. The spatial registration function generates MNI coordinates, anatomical labels, and Brodmann areas maximally represented by each channel.
<img alt="Figure 2" class="xfigimg" s...

Watch this Scientific Journal Video about Neuroimaging Field Methods Using Functional Near Infrared Spectroscopy NIRS Neuroimaging to Study Global Child Development: Rural Sub-Saharan Africa at JoVE.c

Neuroimaging Field Methods Using Functional Near Infrared Spectroscopy NIRS Neuroimaging to Study Global Child Development: Rural Sub-Saharan Africa

Portable neuroimaging approaches (functional Near Infrared Spectroscopy) provide advances to the study of the brain in previously inaccessible regions; here, rural C&#244;te d&#39;Ivoire. Innovation in methods and development of culturally-appropriate neuroimaging protocols permits novel study of the brain&#39;s development and children&#39;s learning outcomes in environments with significant poverty and adversity.

Neuroimaging Field Methods Using Functional Near Infrared Spectroscopy (NIRS) Neuroimaging to Study Global Child Development: Rural Sub-Saharan Africa

Portable neuroimaging approaches provide new advances to the study of brain function and brain development with previously inaccessible populations and in ...

The overall goal of this Field Neuro Imaging Protocol, is to study children's brain development, and cognitive and learning outcomes, using portable functional near infrared spectroscopy, known as FNIRS, in remote understudied, low resource settings such as impoverished rural communities, in Sub-Saharan Africa. This method can help us answer key questions about brain development in high risk environments. For example, how the brain's network for reading develops in communities with high risk of illiteracy.This method also addresses important theoretical issues in global child development, regarding how both biological development and diverse, and often negative experiences, impact the brain. And can reveal new information about developmental, sensitive periods. To begin, invest in the formation of collaborations, and provide opportunities, within the research framework, to local researchers.Establish partnerships with local, scientific institutions for the purpose of inclusivity, as recognition from peers at the local level is important in communicating research findings in the region. Prepare for rainy climate conditions, in the field, as temperature and humidity conditions can vary significantly from laboratory settings, which may effect equipment function and longevity, and participant comfort. Demonstrating the procedure will be Lais Freitas, a visiting scholar in my laboratory, from the Federal University of San Pallo, and Violet Kozloff, our laboratory manager.Begin by undergoing a consent procedure, in a local or preferred language of the participant, that is culturally appropriate, and ensures that participants, their families, and communities are informed about research and their decision to participate in the study. Then, escort the participants to the testing room, and have them sit on a chair. Explain to the participant that a head measurement will be taken.Using a standard tape measure, measure the distances between the nasion and inion around the head, the nasion and inion over the top of the head, through the midline central, or CC, and the distance between the left and right ear tragus, over the top of the head, through CC.Next, place the fNIRS optode holder cap onto the participants head, aligning the cap to the International 10-20 system, for scalp locations. Secure the cap with the strap. Once the cap is positioned, instruct the participant to sit still, while a 3D digitizer measurement of the key 10-20 system scalp positions, and each optode place holder is taken.Then, arrange the 3D digitizer equipment, by placing one sensor on the participant's head, at CC, and affixed securely, and placing the second sensor on the table directly behind the participant's head. Move 3D digitizer stylist to each probe location, and across the key 10-20 system positions. After the 3D digitizer data are collected, direct the participant to be seated comfortably, in front of the stimulus presentation computer.Using the fNIRS built in software, select the probe arrangement that corresponds to the experiment's design. Ensure it is placed to maximally overlay left hemisphere language areas, and their right hemisphere homologues, as well as the frontal lobe. Ensure that both emitter and detector probes are numbered according to the probe arrangement map.Using the optode map and the fNIRS inbuilt software as a guide, place each optode in it's respective opening. Move any hair to ensure contact between the tip of the optode and scalp. Finally, after all of the optodes are in position, check for signal quality, using the fNIRS system built in software.Adjust individual probes, as necessary, until sufficient signal quality is achieved. Begin by placing noise-canceling headphones on the participant's head, being careful not to interfere with fNIRS probe placement. Ensure that the headphones deliver auditory speech stimuli, if applicable to the participant, as well as block any ambient noise.Instruct the participant to fixate on the cross, in the middle of the screen. Ask them to remain still during the experiment, while the experimental tasks are displayed. Dim the lights, and begin recording the participant on the built in video camera.Begin fNIRS data recording, on the fNIRS command computer, and commence tasks on the stimulus presentation computer. Monitor participant performance, throughout all tasks, and provide breaks between tasks and runs. Ensure that triggering, from the experimental stimuli presentation computer is received by the fNIRS command computer.Finally, at the end of all tasks, stop collecting video and fNIRS data. After the experiment is complete, remove each optode from the optode holder cap. Without disrupting the position of the optode holder cap on the participant's head, direct the participant to sit in a position, to obtain a second 3D digitizer measurement.Repeat the 3D digitizer measurement, to ensure that any disruptions to the scalp probe positions during the experiment can be detected, by comparing the two position files. Finally, after the repeat measurement is complete, remove the optode holder cap from the participant's head. The 3D position data and experimental design data, were combined with fNIRS time series data, for analysis in order to map experiment related, significant neural activation patterns on a standard brain template.This subject showed greater activation in the left hemisphere superior temporal gyrus, during rhyming trials compared with rest. Further, greater activation for nonrhyming verses rhyming trials, in the left hemisphere is shown. Such as Subject 1 showed greater activation in the left inferior frontal gyrus, and Subject 2 showed greater activation in the left superior temporal gyrus.Following this procedure, other methods like assessments of cognitive and verbal skills can be performed in order to answer additional questions about the association between brain activity and children's abilities. After watching this video, you should have a good understanding of important considerations in field neuroimaging, such as local partnerships with communities and researchers, as well as a technical understanding of how to position the fNIRS cap, place the optodes, and collect data with this system. Don't forget that working in remote locations can be dangerous, with risks of illness, crime, and political instability.And precautions such as medication, vaccinations, and monitoring travel advisories should always be taken while performing this procedure.

neuroimaging-field-methods-using-functional-near-infrared

Research

JoVE Journal

Neuroscience

Basics of Multivariate Analysis in Neuroimaging Data

Multivariate analysis techniques for neuroimaging data have recently received increasing attention as they have many attractive features that cannot be easily realized by the more commonly used univariate, voxel-wise, techniques1,5,6,7,8,9. Multivariate approaches evaluate correlation/covariance of activation across brain regions, rather than proceeding on a voxel-by-voxel basis. Thus, their results can be more easily interpreted as a signature of neural networks. Univariate approaches, on the other hand, cannot directly address interregional correlation in the brain. Multivariate approaches can also result in greater statistical power when compared with univariate techniques, which are forced to employ very stringent corrections for voxel-wise multiple comparisons. Further, multivariate techniques also lend themselves much better to prospective application of results from the analysis of one dataset to entirely new datasets. Multivariate techniques are thus well placed to provide information about mean differences and correlations with behavior, similarly to univariate approaches, with potentially greater statistical power and better reproducibility checks. In contrast to these advantages is the high barrier of entry to the use of multivariate approaches, preventing more widespread application in the community. To the neuroscientist becoming familiar with multivariate analysis techniques, an initial survey of the field might present a bewildering variety of approaches that, although algorithmically similar, are presented with different emphases, typically by people with mathematics backgrounds. We believe that multivariate analysis techniques have sufficient potential to warrant better dissemination. Researchers should be able to employ them in an informed and accessible manner. The current article is an attempt at a didactic introduction of multivariate techniques for the novice. A conceptual introduction is followed with a very simple application to a diagnostic data set from the Alzheimer s Disease Neuroimaging Initiative (ADNI), clearly demonstrating the superior performance of the multivariate approach.

The current article describes the basics of multivariate analysis and contrasts it to the more commonly used voxel-wise univariate analysis. Both types of analysis are applied to a clinical-neuroscience data set. Supplementary split-half simulations show better replication of the multivariate results in independent data sets.

Multivariate analysis techniques for neuroimaging data have recently received increasing attention as they have many attractive features that cannot be ...

Using MazeSuite and Functional Near Infrared Spectroscopy to Study Learning in Spatial Navigation

MazeSuite is a complete toolset to prepare, present and analyze navigational and spatial experiments1. MazeSuite can be used to design and edit adapted virtual 3D environments, track a participants' behavioral performance within the virtual environment and synchronize with external devices for physiological and neuroimaging measures, including electroencephalogram and eye tracking.
Functional near-infrared spectroscopy (fNIR) is an optical brain imaging technique that enables continuous, noninvasive, and portable monitoring of changes in cerebral blood oxygenation related to human brain functions2-7. Over the last decade fNIR is used to effectively monitor cognitive tasks such as attention, working memory and problem solving7-11. fNIR can be implemented in the form of a wearable and minimally intrusive device; it has the capacity to monitor brain activity in ecologically valid environments. 
Cognitive functions assessed through task performance involve patterns of brain activation of the prefrontal cortex (PFC) that vary from the initial novel task performance, after practice and during retention12. Using positron emission tomography (PET), Van Horn and colleagues found that regional cerebral blood flow was activated in the right frontal lobe during the encoding (i.e., initial na&iuml;ve performance) of spatial navigation of virtual mazes while there was little to no activation of the frontal regions after practice and during retention tests. Furthermore, the effects of contextual interference, a learning phenomenon related to organization of practice, are evident when individuals acquire multiple tasks under different practice schedules13,14. High contextual interference (random practice schedule) is created when the tasks to be learned are presented in a non-sequential, unpredictable order. Low contextual interference (blocked practice schedule) is created when the tasks to be learned are presented in a predictable order.
Our goal here is twofold: first to illustrate the experimental protocol design process and the use of MazeSuite, and second, to demonstrate the setup and deployment of the fNIR brain activity monitoring system using Cognitive Optical Brain Imaging (COBI) Studio software15. To illustrate our goals, a subsample from a study is reported to show the use of both MazeSuite and COBI Studio in a single experiment. The study involves the assessment of cognitive activity of the PFC during the acquisition and learning of computer maze tasks for blocked and random orders. Two right-handed adults (one male, one female) performed 315 acquisition, 30 retention and 20 transfer trials across four days. Design, implementation, data acquisition and analysis phases of the study were explained with the intention to provide a guideline for future studies.

MazeSuite is a complete toolset to prepare, present and analyze navigational and spatial experiments. Functional near-infrared spectroscopy (fNIR) is an optical brain imaging technique that enables noninvasive and portable monitoring of cerebral blood oxygenation changes. This paper summarizes collective use of MazeSuite and fNIR within a cognitive processing learning paradigm.

MazeSuite is a complete toolset to prepare, present and analyze navigational and spatial experiments1. MazeSuite can be used to design and edit adapted ...

Local and Global Methods of Assessing Thermal Nociception in Drosophila Larvae

In this article, we demonstrate assays to study thermal nociception in Drosophila larvae. One assay involves spatially-restricted (local) stimulation of thermal nociceptors1,2 while the second involves a wholesale (global) activation of most or all such neurons3. Together, these techniques allow visualization and quantification of the behavioral functions of Drosophila nociceptive sensory neurons.
The Drosophila larva is an established model system to study thermal nociception, a sensory response to potentially harmful temperatures that is evolutionarily conserved across species1,2. The advantages of Drosophila for such studies are the relative simplicity of its nervous system and the sophistication of the genetic techniques that can be used to dissect the molecular basis of the underlying biology4-6 In Drosophila, as in all metazoans, the response to noxious thermal stimuli generally involves a "nocifensive" aversive withdrawal to the presented stimulus7. Such stimuli are detected through free nerve endings or nociceptors and the amplitude of the organismal response depends on the number of nociceptors receiving the noxious stimulus8. In Drosophila, it is the class IV dendritic arborization sensory neurons that detect noxious thermal and mechanical stimuli9 in addition to their recently discovered role as photoreceptors10. These neurons, which have been very well studied at the developmental level, arborize over the barrier epidermal sheet and make contacts with nearly all epidermal cells11,12. The single axon of each class IV neuron projects into the ventral nerve cord of the central nervous system11 where they may connect to second-order neurons that project to the brain.
Under baseline conditions, nociceptive sensory neurons will not fire until a relatively high threshold is reached. The assays described here allow the investigator to quantify baseline behavioral responses or, presumably, the sensitization that ensues following tissue damage. Each assay provokes distinct but related locomotory behavioral responses to noxious thermal stimuli and permits the researcher to visualize and quantify various aspects of thermal nociception in Drosophila larvae. The assays can be applied to larvae of desired genotypes or to larvae raised under different environmental conditions that might impact nociception. Since thermal nociception is conserved across species, the findings gleaned from genetic dissection in Drosophila will likely inform our understanding of thermal nociception in other species, including vertebrates.

Local and Global Methods of Assessing Thermal Nociception in Drosophila Larvae

In this article, we demonstrate assays to study thermal nociception in Drosophila larvae. One assay involves spatially-restricted (local) stimulation of thermal nociceptors1,2 while the second involves a wholesale (global) activation of most or all such neurons3. Together, these techniques allow visualization and quantification of the behavioral functions of Drosophila nociceptive sensory neurons.

In this article, we demonstrate assays to study thermal nociception in Drosophila larvae. One assay involves spatially-restricted (local) stimulation of ...

Functional Neuroimaging Using Ultrasonic Blood-brain Barrier Disruption and Manganese-enhanced MRI

Although mice are the dominant model system for studying the genetic and molecular underpinnings of neuroscience, functional neuroimaging in mice remains technically challenging. One approach, Activation-Induced Manganese-enhanced MRI (AIM MRI), has been used successfully to map neuronal activity in rodents 1-5. In AIM MRI, Mn2+ acts a calcium analog and accumulates in depolarized neurons 6,7. Because Mn2+ shortens the T1 tissue property, regions of elevated neuronal activity will enhance in MRI. Furthermore, Mn2+ clears slowly from the activated regions; therefore, stimulation can be performed outside the magnet prior to imaging, enabling greater experimental flexibility. However, because Mn2+ does not readily cross the blood-brain barrier (BBB), the need to open the BBB has limited the use of AIM MRI, especially in mice.
One tool for opening the BBB is ultrasound. Though potentially damaging, if ultrasound is administered in combination with gas-filled microbubbles (i.e., ultrasound contrast agents), the acoustic pressure required for BBB opening is considerably lower. This combination of ultrasound and microbubbles can be used to reliably open the BBB without causing tissue damage 8-11.
Here, a method is presented for performing AIM MRI by using microbubbles and ultrasound to open the BBB. After an intravenous injection of perflutren microbubbles, an unfocused pulsed ultrasound beam is applied to the shaved mouse head for 3 minutes. For simplicity, we refer to this technique of BBB Opening with Microbubbles and UltraSound as BOMUS 12. Using BOMUS to open the BBB throughout both cerebral hemispheres, manganese is administered to the whole mouse brain. After experimental stimulation of the lightly sedated mice, AIM MRI is used to map the neuronal response.
To demonstrate this approach, herein BOMUS and AIM MRI are used to map unilateral mechanical stimulation of the vibrissae in lightly sedated mice 13. Because BOMUS can open the BBB throughout both hemispheres, the unstimulated side of the brain is used to control for nonspecific background stimulation. The resultant 3D activation map agrees well with published representations of the vibrissae regions of the barrel field cortex 14. The ultrasonic opening of the BBB is fast, noninvasive, and reversible; and thus this approach is suitable for high-throughput and/or longitudinal studies in awake mice.

A technique is described for broadly opening the blood-brain barrier in the mouse using microbubbles and ultrasound. Using this technique, manganese can be administered to the mouse brain. Because manganese is an MRI contrast agent that accumulates in depolarized neurons, this approach enables imaging of neuronal activity.

Although mice are the dominant model system for studying the genetic and molecular underpinnings of neuroscience, functional neuroimaging in mice remains ...

New Methods to Study Gustatory Coding

The sense of taste allows animals to detect chemicals in the environment, giving rise to behaviors critical for survival. When Gustatory Receptor Neurons (GRNs) detect tastant molecules, they encode information about the identity and concentration of the tastant as patterns of electrical activity that then propagate to follower neurons in the brain. These patterns constitute internal representations of the tastant, which then allow the animal to select actions and form memories. The use of relatively simple animal models has been a powerful tool to study basic principles in sensory coding. Here, we propose three new methods to study gustatory coding using the moth Manduca sexta. First, we present a dissection procedure for exposing the maxillary nerves and the subesophageal zone (SEZ), allowing recording of the activity of GRNs from their axons. Second, we describe the use of extracellular electrodes to record the activity of multiple GRNs by placing tetrode wires directly into the maxillary nerve. Third, we present a new system for delivering and monitoring, with high temporal precision, pulses of different tastants. These methods allow the characterization of neuronal responses in vivo directly from GRNs before, during and after tastants are delivered. We provide examples of voltage traces recorded from multiple GRNs, and present an example of how a spike sorting technique can be applied to the data to identify the responses of individual neurons. Finally, to validate our recording approach, we compare extracellular recordings obtained from GRNs with tetrodes to intracellular recordings obtained with sharp glass electrodes.

We present three new methods to study gustatory coding. Using a simple animal, the moth Manduca sexta (Manduca), we describe a dissection protocol, the use of extracellular tetrodes to record the activity of multiple gustatory receptor neurons, and a system for delivering and monitoring precisely timed pulses of tastants.

The sense of taste allows animals to detect chemicals in the environment, giving rise to behaviors critical for survival. When Gustatory Receptor Neurons ...

Conducting Hyperscanning Experiments with Functional Near-Infrared Spectroscopy

Concurrent brain recordings of two or more interacting persons, an approach termed hyperscanning, are gaining increasing importance for our understanding of the neurobiological underpinnings of social interactions, and possibly interpersonal relationships. Functional near-infrared spectroscopy (fNIRS) is well suited for conducting hyperscanning experiments because it measures local hemodynamic effects with a high sampling rate and, importantly, it can be applied in natural settings, not requiring strict motion restrictions. In this article, we present a protocol for conducting fNIRS hyperscanning experiments with parent-child dyads and for analyzing brain-to-brain synchrony. Furthermore, we discuss critical issues and future directions, regarding the experimental design, spatial registration of the fNIRS channels, physiological influences and data analysis methods. The described protocol is not specific to parent-child dyads, but can be applied to a variety of different dyadic constellations, such as adult strangers, romantic partners or siblings. To conclude, fNIRS hyperscanning has the potential to yield new insights into the dynamics of the ongoing social interaction, which possibly go beyond what can be studied by examining the activities of individual brains.

The present protocol describes how to conduct fNIRS hyperscanning experiments and analyze brain-to-brain synchrony. Further, we discuss challenges and possible solutions.

Concurrent brain recordings of two or more interacting persons, an approach termed hyperscanning, are gaining increasing importance for our understanding ...

Conducting Concurrent Electroencephalography and Functional Near-Infrared Spectroscopy Recordings with a Flanker Task

Concurrent EEG and fNIRS recordings offer an excellent opportunity to gain a full understanding of the neural mechanism of cognitive processing by inspecting the relationship between the neural and hemodynamic signals. EEG is an electrophysiological technology that can measure the rapid neuronal activity of the cortex, whereas fNIRS relies on the hemodynamic responses to infer brain activation. The combination of EEG and fNIRS neuroimaging techniques can identify more features and reveal more information associated with the functioning of the brain. In this protocol, fused EEG-fNIRS measurements were performed for concurrent recordings of evoked-electrical potentials and hemodynamic responses during a Flanker task. In addition, the critical steps for setting up the hardware and software system as well as the procedures for data acquisition and analysis were provided and discussed in detail. It is expected that the present protocol can pave a new avenue for improving the understanding of the neural mechanisms underlying various cognitive processes by using the EEG and fNIRS signals.

The present protocol describes how to perform concurrent EEG and fNIRS recordings and how to inspect the relationship between the EEG and fNIRS data.&#160;

Concurrent EEG and fNIRS recordings offer an excellent opportunity to gain a full understanding of the neural mechanism of cognitive processing by ...

Role of Diffusion MRI Tractography in Endoscopic Endonasal Skull Base Surgery

Endoscopic endonasal surgery has gained a prominent role in the management of complex skull base tumors. It allows the resection of a large group of benign and malignant lesions through a natural anatomical extra-cranial pathway, represented by the nasal cavities, avoiding brain retraction and neurovascular manipulation. This is reflected by the patients' prompt clinical recovery and the low risk of permanent neurological sequelae, representing the main caveat of conventional skull base surgery. This surgery must be tailored to each specific case, considering its features and relationship with surrounding neural structures, mostly based on preoperative neuroimaging. Advanced MRI techniques, such as tractography, have been rarely adopted in skull base surgery due to technical issues: lengthy and complicated processes to generate reliable reconstructions for inclusion in the neuronavigation system. 
This paper aims to present the protocol implemented in the institution and highlights the synergistic collaboration and teamwork between neurosurgeons and the neuroimaging team (neurologists, neuroradiologists, neuropsychologists, physicists, and bioengineers) with the final goal of selecting the optimal treatment for each patient, improving the surgical results and pursuing the advancement of personalized medicine in this field.

We present a protocol to integrate diffusion MRI tractography in patient work-up to endoscopic endonasal surgery for a skull base tumor. The methods for adopting these neuroimaging studies in the pre- and intra-operative phases are described.

Endoscopic endonasal surgery has gained a prominent role in the management of complex skull base tumors. It allows the resection of a large group of ...

Qualitative and Comparative Cortical Activity Data Analyses from a Functional Near-Infrared Spectroscopy Experiment Applying Block Design

Neuroimaging studies play a pivotal role in the evaluation of pre- vs. post-interventional neurological conditions such as in rehabilitation and surgical treatment. Among the many neuroimaging technologies used to measure brain activity, functional near-infrared spectroscopy (fNIRS) enables the evaluation of dynamic cortical activities by measuring the local hemoglobin levels similar to functional magnetic resonance imaging (fMRI). Also, due to lesser physical restriction in fNIRS, multiple variants of sensorimotor tasks can be evaluated. Many laboratories have developed several methods for fNIRS data analysis; however, despite the fact that the general principles are the same, there is no universally standardized method. Here, we present the qualitative and comparative analytic methods of data obtained from a multi-channel fNIRS experiment using a block design. For qualitative analysis, we used a software for NIRS as a mass-univariate approach based on the generalized linear model. The NIRS-SPM analysis shows qualitative results for each session by visualizing the activated area during the task. In addition, the non-invasive three-dimensional digitizer can be used to estimate the fNIRS channel locations relative to the brain. To corroborate the NIRS-SPM findings, the amplitude of the changes in hemoglobin levels induced by the sensorimotor task can be statistically analyzed by comparing the data obtained from two different sessions (before and after intervention) of the same study subject using a multi-channel hierarchical mixed model. Our methods can be used to measure the pre- vs. post-intervention analysis in a variety of neurological disorders such as movement disorders, cerebrovascular diseases, and neuropsychiatric disorders.

We describe the analysis of continuous-wave functional near-infrared spectroscopy experiment using a block design with a sensorimotor task. To increase the reliability of the data analysis, we used the qualitative general linear model-based statistical parametric mapping and the comparative hierarchical mixed models for multi-channels.

Neuroimaging studies play a pivotal role in the evaluation of pre- vs. post-interventional neurological conditions such as in rehabilitation and surgical ...

Employing Transcranial Magnetic Stimulation in a Resource Limited Environment to Establish Brain-Behavior Relationships

Neuroimaging is typically perceived as a resource demanding discipline. While this is the case in certain circumstances, institutions with limited resources have historically contributed significantly to the field of neuroscience, including neuroimaging. In the study of self-deception, we have successfully employed single-pulse TMS to determine the brain correlates of abilities including overclaiming and self-enhancement. Even without the use of neuro-navigation, methods provided here lead to successful outcomes. For example, it was discovered that decreases in self-deceptive responding lead to a decrease in affect. These methods provide data that are reliable and valid, and such methods provide research opportunities otherwise unavailable.&#160;Through the use of these methods, the overall knowledge base in the field of neuroscience is expanded, providing research opportunities to students such as those at our institution&#160;(Montclair State University is a Hispanic-Serving Institute) who are often denied such research experiences.

Transcranial Magnetic Stimulation (TMS) and low-frequency TMS (lfTMS) have&#160;been demonstrated to be major contributors to brain literature. Here we highlight the methods for investigating the cortical correlates of self-deception using TMS.