Portable neuroimaging approaches (functional Near Infrared Spectroscopy) provide advances to the study of the brain in previously inaccessible regions; here, rural Côte d'Ivoire. Innovation in methods and development of culturally-appropriate neuroimaging protocols permits novel study of the brain's development and children's learning outcomes in environments with significant poverty and adversity.
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 Côte d'Ivoire. Innovation in methods and the development of culturally appropriate neuroimaging protocols allow a first-time look into the brain's development and children'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 Côte d'Ivoire, the world'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.
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—including the powerful effects of a child's environment and their early-life experiences1,2,3. Methods that advance the field'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 Côte d'Ivoire. The aim of this field neuroimaging research program was to understand children's reading development in an environment with a high-risk of illiteracy. Côte d'Ivoire's youth (15-24 years) literacy rate is 53%, despite 93% primary school enrollment rates4. Côte d'Ivoire is the world'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, can yield valuable insights into children’s learning outcomes. 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's learning outcomes.
fNIRS neuroimaging is well-suited for field research. Similar to functional magnetic resonance imaging (fMRI), fNIRS measures the brain'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' 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 Côte d'Ivoire, the protocol is highly relevant for any field neuroimaging study in challenging, low-resource environments, and can be adapted for novel contexts.
All methods described here have been approved by the Institutional Review Board (IRB) of the University of Delaware.
1. Mobile Laboratory Transport and Setup
Figure 1. Schematics. (A) Schematic of laboratory setup. (B) Preparing the participant for data collection. Please click here to view a larger version of this figure.
2. Local Research Teams and Science Partners
3. Informed Consent and Child Assent
4. fNIRS Scalp Placement and Measurement
5. Experimental Tasks
6. Post-experimental Task Measurements
7. Plan for Disseminating Data
8. Backup Data
9. Data Analysis
NOTE: Multiple data analysis packages exist for fNIRS14. Statistical Parametric Mapping for Near-Infrared Spectroscopy (NIRS-SPM)15 , Homer216 (widely used), and the fNIRS toolbox 17,18 (new and gaining popularity) are used for fNIRS data analysis. This protocol reviews data analysis methods using NIRS-SPM, but it is to the discretion of the researcher to select preferred method of analysis.
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's stand-alone registration function25. The spatial registration function generates MNI coordinates, anatomical labels, and Brodmann areas maximally represented by each channel.
Figure 2. Data collection. (A) Placement of the fNIRS cap on the participant's head and collection of position data using the 3D digitizer. (B) International 10-20 system used to guide placement of the cap on the participant's head. (C) Spatial localization algorithm plotting x, y, z coordinate data on the MNI brain template. The image generated during stand-alone NIRS registration using 3D digitizer data in NIRS-SPM15,19,25. Please click here to view a larger version of this figure.
Probe position data can also be visualized over cortex surface template or anatomical MRI template using Brainstorm software (Figure 3).
Figure 3. fNIRS probes. fNIRS probes visualized on (A) the surface of the cortex and (B) the MNI anatomical MRI template. Left, dorsal, and right views are presented. Images generated using Brainstorm software13. Please click here to view a larger version of this figure.
Here, representative data from the rhyme judgment task are shown (Figure 4). Participants completed two identically-structured runs of this task. Each run contained 13 trials; rhyming and non-rhyming trials were randomly ordered.
Figure 4. Task design. The rhyme judgment task scheme is shown. Participants continuously viewed a fixation cross while periodically listening to French rhyming or non-rhyming word pairs. The task was completed in two runs, each comprised of 13 trials. 13 rhyming and 13 non-rhyming trials were randomly presented. Each trial lasted 3 s; 1 s per word with a 1 s ISI. Jittered presentation of rest periods between trials, which lasted 8-17 s. Please click here to view a larger version of this figure.
The 3D position data and experimental design data were combined with fNIRS time-series data (Figure 5) for analysis in order to map experiment-related significant neural activation patterns on a standard brain template (Figure 6). Representative single subject data and results are shown in Figure 5 and Figure 6.
Figure 5. Representative time-series data from one fNIRS channel. (A) Raw time-series data corresponding to entire task length (rhyme judgment task; first run), not normalized. (B) Filtered time-series data using wavelet detrending. Rhyming and non-rhyming trials are indicated by solid and dashed box-car, respectively. Please click here to view a larger version of this figure.
This subject showed greater activation in the left hemisphere Superior Temporal Gyrus (STG) during rhyming trials compared with rest (baseline fixation cross). Averaged HbO and HbR responses for rhyming trials show a canonical hemodynamic response: increasing HbO concentrations and corresponding decreasing HbR concentrations following stimulus presentation.
Figure 6. Representative single-subject results. (A) Greater activation is observed for rhyming trials versus baseline (fixation cross) in the left hemisphere, overlaying the posterior portion of the superior temporal gyrus (STG). Image generated during NIRS results steps using NIRS-SPM15,19,25. (B) Averaged event-related waveforms for HbO (red) and HbR (blue) during rhyming trials (rhyming word pair stimuli). Image generated using Xu Cui's plot average function26. Please click here to view a larger version of this figure.
Single-subject results differed between participants (see Figure 7). This individual variation may reflect underlying functional differences or developmental differences in the organization of specific brain networks. For example, subject 1 showed greater activation in the left inferior frontal gyrus region during non-rhyming versus rhyming trials; whereas subject 2 showed greater activation in the left STG region during the same experimental contrast (non-rhyming versus rhyming trials).
Figure 7. Representative single-subject results from two different participants for identical contrast. Greater activation for non-rhyming versus rhyming trials in the left hemisphere is shown in both A and B. (A) Subject 1 showed greater activation in the left inferior frontal gyrus. (B) Subject 2 showed greater activation the left superior temporal gyrus. Please click here to view a larger version of this figure.
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 strong partnerships with local scientific, educational, and government institutions, and we highlight the reciprocal knowledge transfer that occurs when successful long-term partnerships are formed between local and visiting scientists. Guidelines for the development of culturally-appropriate informed consent procedures and testing protocols are discussed with the aim of incorporating multiple cultural perspectives in research methods. Finally, this protocol provides detailed steps for field data collection and data analysis.
Local Science Engagement and Opportunities for Capacity Building:
One of the main challenges that local, particularly junior, researchers in Côte d'Ivoire are faced with when they complete their studies is the lack of opportunity for hands-on research experience with experienced researcher mentors and/or international collaborators. For this purpose, researchers should make all efforts to establish robust collaboration with local researchers from relevant disciplinary backgrounds, and include trainees at all levels (undergraduate, graduate, postdoctoral). Trainees can leverage the insight gained from this experience to work independently and further research. This experience can also be a stepping stone to build their capacities as researchers and develop their competitiveness at the international level in writing research proposals and papers and applying for grants. A research team excluding local researchers may have a reduced chance of success as local researchers will best know the local social and cultural values and systems, the local languages spoken in addition to the geographic knowledge of the area. Their contribution is therefore extremely important in understanding the local realities and designing culturally-appropriate protocols for the successful research projects.
Culturally-appropriate Research Methods:
The development of informed consent protocols to conduct research in rural settings specifically in Cote d'Ivoire is critical and failure to adopt the appropriate approach can inhibit the successful achievement of the research even though well-intentioned and scientifically robust27,28,29,30,31. Generally, in rural settings in Côte d'Ivoire, asking a villager to read a consent form and sign it can break any trust building between the researcher and the participant. In fact, the perceived formality of this procedure may create a psychological distance and a feeling of insecurity in the participant's mind. This may result in a clear or unexpressed unwillingness to collaborate. This attitude can be explained by many factors including a long history of oral tradition whereby communication is more oral than written and high rates of illiteracy that may be found in target communities. Communities in rural settings trust their chief and rely on his decision-making power. Therefore, the protocol presented incorporates the consent of the chief of the village at the community level. This is arguably more culturally important than individual consent. Additionally, participants and community members may have had limited or no exposure to neuroimaging technology or computers. Therefore, researchers need to take into consideration that the informed consent procedure, and instructions, may be misunderstood. The function of the fNIRS system should be communicated in lay terms and appropriate language easily understandable by child participants and community members who may have had very limited exposure to technology. These considerations can strongly influence the comfort and confidence of all community members involved in a field neuroimaging research project.
The protocol presented here also highlighted the importance of sharing research findings with community members and government partners. Partnerships built on continued dialogue aid in the eventual translation of research findings into policy. It is imperative to arrange post data collection field visits to disseminate research findings and deliver reports and, possibly, share any tools that resulted from the study (e.g., assessments in local languages). Participating communities in rural settings may never otherwise receive information about study completion and findings given lack of internet service and/or computers. Likewise, researchers in the country may have limited access to academic journal subscriptions and poor internet connectivity at regional universities. Published results should be shared in a regional forum, and made available in an accessible language.
Limitations and Potential Challenges:
This field neuroimaging protocol should be modified to suit the planned data collection sites. The protocol presented here has been developed for research with primary school aged children in rural Cote d'Ivoire. However, the methods outlined here may not be suitable, specifically with respect to informed consent procedures, in other countries or even other regions of Cote d'Ivoire. Researchers who aim to conduct field neuroimaging must first carefully research local customs and incorporate local perspectives into study design. Therefore, a research team working on study designs must include members from the local cultural groups.
Field neuroimaging has limitation in comparison with laboratory methods. Importantly, control of the testing environment is considerably reduced in the field. Field neuroimaging researchers should plan extended data collection trips. Tropical rains, risk of contracting tropical diseases, civil strikes, and political unrest may significantly impact research plans. Researchers need to ensure security levels in the region are sufficient and monitor for updates to any situations that may affect security levels. Continuous communication between team members, specifically with respect to security levels, may mitigate potential risks.
Future Applications and Relevance to Existing Methods:
The use of this field neuroimaging method can be applied to evaluating the impact of early risk on infant and child development in global health settings. Researchers have begun using this approach to study child development in rural Gambia and an urban slum in Bangladesh32. In an urban slum in Dhaka, researchers are using fNIRS to examine how factors such as nutrition and sanitation contribute to growth and brain development33. In rural Gambia, researchers have used fNIRS to study cognitive function of infants, and have demonstrated that fNIRS is a viable imaging tool in resource-poor settings34,35. Such work promises to reveal new insights into the development of children in the developing world, who are disproportionately affected by malnutrition and poor sanitation. Portable neuroimaging technologies continue to become more accessible and applicable for research in low-resource environments, thus highlighting the need for rigorous methods for field neuroimaging.
Conclusion:
Portable neuroimaging systems with the capability to function on battery-supplied power have recently become available. As these techniques are relatively new, advances to battery systems will provide ongoing improvements. Diverse communities of international scientists developing research programs using these tools will undoubtedly optimize mobile laboratory settings to provide increased control of the testing environment. Meaningful engagement between international and local scientists and local communities can ensure that members of study populations have active roles in the development of research programs and represent the interests of their communities. Only such collaborative and integrated research teams would be well-positioned to apply field neuroimaging methods to study all human brain development, and reveal both theoretically- and practically-relevant information aimed at understanding the most urgent child development issues.
The authors have nothing to disclose.
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 Moapé, Ananguié, Affery, and Becouefin for their participation in this research program and the villages' warm hospitality.
LIGHTNIRS Main Unit Pack 120V | Shimadzu | 292-34000-42 | Component of the fNIRS system |
HOLDER ASSY, ALL- CAP | Shimadzu | 594-07618-01 | Component of the fNIRS system |
LIGHTNIRS connection cable | Shimadzu | 567-10976-11 | fNIRS system component |
Fiber set for LIGHTNIRS, 1m (8 sets) | Shimadzu | 567-11350-01 | fNIRS system component |
Dell Latitude Laptop | Shimadzu (from Dell) | 220-97322-00 | Master computer to run fNIRS applications |
PATRIOT SEU (System Electronics Unit) | POLHEMUS | 1A0453-001 | PATRIOT System component |
Power Supply | POLHEMUS | 2C0809 | PATRIOT System component |
Power Supply cord | POLHEMUS | 17500B-BLK | PATRIOT System component |
RS-232 null modem cable | POLHEMUS | 1C0288 | PATRIOT System component |
USB cable | POLHEMUS | 1C0289 | PATRIOT System component |
RX2 Sensor 10' cable | POLHEMUS | 4A0492-20 | PATRIOT System component |
TX2 Source 10' cable | POLHEMUS | 4A0506-20 | PATRIOT System component |