Testing and Calibration of Non-Invasive Optical Imaging Technology for Functional Brain Imaging
Phase: NA
Conditions: Cognition
Sponsor: Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD)

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 0 of 28
Abbreviated Title: Non-Invasive Optical Imaging
CC Protocol #: 10-CH-0198
NCT Number: NCT01212029
Version Date: 04/01/2022
Title: Testing and Calibration of Non-invasive Optical Imaging Technology for 
Functional Brain Imaging
NIH Principal Investigator:
Amir Gandjbakhche, PhD
National Institute for Child Health and Human 
Development (NICHD)
Building 49, Room 5A82
49 Convent Drive
Bethesda, MD 20892
Phone: 301-435-9235
E-mail: gandjbaa@mail.nih.gov 

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 1 of 28
Précis:
Objective: to a) cross-validate our near infrared spectroscopy (NIRS) imaging system with 
existing functional magnetic resonance imaging (fMRI) and electroencephalogram (EEG) 
data, and b) to investigate any significant technical issues associated with optode placement 
and motion artifacts, and to explore techniques that will potentially improve the feasibility 
and reliability of the system according to the needs of the population for whom existing 
imaging systems are unsuitable.
Study population: 250 healthy volunteers
Design: The study will look for correlations between NIRS signal changes in healthy 
subjects when performing functional tasks, and existing fMRI data. 
Outcome Measures: graded changes in blood flow and oxygen, measured with NIRS, in 
response to different functional tasks.

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 2 of 28
Contents
Précis: .........................................................................................................................................................1
List of Abbreviations.............................................................................................................................3
1
Introduction and Background .....................................................................................................3
2
Study Objectives .............................................................................................................................5
3
Subjects.............................................................................................................................................6
4
Study Design and Methods ...........................................................................................................6
5
Management of Data and Samples...........................................................................................15
6
Additional Considerations..........................................................................................................16
7
Risks and Discomforts.................................................................................................................17
8
Subject Safety Monitoring..........................................................................................................17
9
Outcome Measures.......................................................................................................................18
10
Statistical Analysis .......................................................................................................................18
11
Human Subjects Protection .......................................................................................................18
12
Anticipated Benefit ......................................................................................................................19
13
Consent Documents and Process ..............................................................................................19
14
Data and Safety Monitoring.......................................................................................................20
15
Quality Assurance........................................................................................................................20
16
NIH Reporting Requirements ...................................................................................................20
16.1
Reportable events definitions .............................................................................................................20
16.2
Expedited reporting................................................................................................................................20
16.3
IRB Requirements for PI Reporting at Continuing Review.................................................20
16.4
Reporting to Clinical Director (CD)................................................................................................21
17
Alternatives to Participation......................................................................................................21
18
Privacy............................................................................................................................................21
19
Confidentiality ..............................................................................................................................21
20
Conflict of Interest .......................................................................................................................21
21
Technology Transfer ...................................................................................................................21
22
Research and Travel Compensation........................................................................................22
23
References......................................................................................................................................23
24
Attachments/ Appendices...........................................................................................................27

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 3 of 28
List of Abbreviations 
NIRS: near-infrared spectroscopy, fMRI: functional magnetic resonance imaging, PET: 
positron emission tomography
1
Introduction and Background
Near-infrared spectroscopy (NIRS) is an emerging technology for noninvasive 
measurement of local changes in cerebral hemodynamic activity using infrared light in 
the range of 700 to 1000 nm (Villringer and Chance, 1997, Yodh et al., 2003, Boes et al. 
2004, Gratton et al. 2005, Huppert et al. 2009). Compared to other well-established brain 
imaging modalities, such as functional magnetic resonance imaging (fMRI) and positron 
emission tomography (PET), this technique has a higher temporal resolution (in order of 
milliseconds), and provides additional spectroscopic information about changes in the 
oxygenation content of cerebral blood. Since this technology measures localized 
hemodynamic changes associated with brain activity, it is considered a measure of 
functional neural activation over time and thus is interchangeably referred to as both 
NIRS and functional NIRS (fNIRS).
The application of NIRS technology for neuroimaging relies on the principle that active 
neural tissues demand and consume more oxygen. The relative increase in oxygenated 
hemoglobin in active brain tissues is called the hemodynamic response. Like fMRI, NIRS 
imaging is a non-invasive technique which measures the relative changes in the 
oxygenation content of hemoglobin in neuronal tissue. However, NIRS utilizes changes 
in the absorption optical spectrum of oxygenated versus deoxygenated hemoglobin rather 
than their magnetic properties. Because of this difference, NIRS imaging makes 
functional brain studies possible for sensitive populations who cannot safely undergo 
MRI, such as hemispherectomy patients, individuals with specific types of implanted 
medical devices, or war veterans with embedded ferromagnetic shrapnel associated with 
a penetrating Traumatic Brain Injury (TBI). NIR instruments are also smaller in size, 
relatively less restraining, and more tolerant of subject motion compared to fMRI or PET, 
making them appealing tools for functional brain imaging studies with infants and 
children (Zaramella et al, 2001, Elwell et al, 2005, Sowell et al, 2004, Nagamitsu et al, 
2006, Wilcox et al, 2005). Validating the use of NIRS as a brain imaging modality is 
crucial for serving populations in which MRI, fMRI, or PET environments are unsuitable.
In a typical brain imaging experiment using fNIRS, an array of detectors and low power 
NIR light sources are placed on the head, orthogonal to the scalp. Due to the low optical 
absorption of biological tissue at NIR wavelengths, NIR light can penetrate  the head skin 
and skull (approximately 1.5-2 cm) to reach the outer5–10mm of the brain tissue surface. 
(Fukui et al, 2003, Okada et al 2003, Huppert et al, 2009). Light entering the head at a 
source and exiting the head at a detector samples a diffuse volume of tissue between these 
source and detector positions (Huppert et al, 2009). Changes in the optical absorption of 
this tissue, represented as changes in the amount of detected light, can be related to 
alteration in the oxyhemoglobin and deoxyhemoglobin concentrations in cerebral blood. 
By measuring changes in the intensity of detected light at distinct wavelengths over time, 

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 4 of 28
functionally evoked changes in hemodynamic activity can be captured and used for 
imaging brain activity in cortical areas.
Like other neuroimaging modalities, NIRS technique has its own unique limitations, 
including limited optimal spatial resolution (approximately 5mm) and limited effective 
brain imaging depth (approximately 3 cm). Specific issues concerning the use of optical 
technology for bioimaging and reconstruction of tomographic images, such as dealing with 
motion artifacts and cross talk, motivate researchers to pursue further investigation on the 
design and application of NIRS imaging. While the use of NIRS technology for functional 
brain imaging has been studied over the past three decades, its involvement in clinical 
research remains extremely limited. Thus, this research intends to address critical questions 
regarding the application of NIRS in clinical settings. Specifically, we intend to address 
the following questions concerning the validation and methodologically testing of NIRS 
neuroimaging.
First, we intend to validate the use of a basic NIR functional imaging (NIRFI) system for 
neuroimaging by comparing NIRFI data to that which has been obtained through fMRI and 
electroencephalogram (EEG) studies. Previous research has compared the study of the 
cerebral hemodynamic response to physiological activation using both blood-oxygen-
level-dependent imaging and arterial spin labeling (ASL) MRI techniques. These studies 
have demonstrated that while the spatial and temporal measures are concordant, there are 
minor differences in latency between the onset of activation and NIRS and MRI measures 
(Kleinschmidt, Obrig et al. 1996, Strangman, Culver et al. 2002, Hoge, Franceschini et al. 
2005, Huppert, Hoge et al. 2006). Using two types of NIRFI devices, fNIRSoft from 
BIOPAC Systems and NIRScout from NIRx, we will measure patterns of functional 
hemodynamic activity in healthy volunteers while they perform tasks which have been 
well-studied and reported in fMRI. Ultimately, we will compare our findings with 
corresponding fMRI results to form a benchmark for validation and calibration our fNIR 
system in assessing functional cerebral activity. As standard NIRFI devices, the light 
sources of both the fNIRSoft and NIRScout systems apply light intensities less than 100 
mW, well below the Federal Drug Administration’s (FDA) eye safety requirements for 
Class I devices. They are passive, non-invasive tools for the measurement of NIRFI data 
and do not require further FDA approval. The specifications of both devices are detailed in 
a subsequent section of this protocol. 
Additionally, electroencephalogram (EEG) measurement of cortical neural activity may be 
performed for the purposes of validating NIRS measurements as well as enhancing their 
temporal resolution. Specifically, we will use the actiCAP (Brain Products LLC), a 32-
channel electrode array, which is designed for use in combination with a variable number 
of NIRS source-detector arrangements. Like both the fNIR Devices and NIRScout devices, 
this is a safe, passive, and noninvasive method of measuring cortical brain activity which 
is commonly used for the functional assessment of coordinated neural activity. The 
specifications of this device are detailed in a subsequent section of this protocol. Utilization 
of these compatibly designed neuroimaging systems will be important for accomplishing 
the study objective to cross-validate the use of near infrared spectroscopy (NIRS) with 
other existing functional methodologies.

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 5 of 28
During functional task experiments, subjects will be fitted with the fNIR device including 
a specific array of sources and detectors. For each task, the subject will be given clear 
instructions before being presented with the task stimulus. As the subject completes each 
specified task, the NIRFI system will continuously record the light intensity data which 
corresponds to changes in oxyhemoglobin and deoxyhemoglobin concentrations in the 
brain tissues. We will utilize systems such as E-Prime to record and correspond the timing 
of stimulus presentation with the measured hemodynamic response. Each functional task 
will have its own base line task designed as a control level, lasting a duration sufficient for 
the resumption of resting state hemodynamic activity before the presentation of any 
subsequent stimuli. Examples and detailed descriptions of the functional tasks which may 
be used are provided in Section 3 of this protocol.
In testing, we will compare block and event related designs. We will also study the impacts 
of varying inter stimulus intervals and repetition rates of the stimulus. Once light intensity 
data is collected from the detectors, principal component analysis (PCA), and independent 
component analysis (ICA) techniques will be applied to remove the noise and motion 
artifact components and obtain the corresponding hemodynamic response data. 
Second, we intend to test the effect of subject behaviors and physiology on the collection 
of NIRFI data. Specifically, we aim to study factors which may confound the collection of 
representative NIRS data, such as the motion of the subject during testing. As in other 
neuroimaging modalities, motion during NIRFI can generate noise in the collected data 
and is considered a confounding issue and limitation. While NIRS imaging techniques are 
more tolerant of noise than fMRI or PET, understanding how motion affects the quality of 
NIRFI data is critical to validation of the technique. In this study, we will evaluate the 
efficacy of various techniques, such as the use of arrays of interlaced electronic sensors, in 
maximizing the system tolerance for motion and minimizing the confound of motion-
related noise in NIRFI data. This will assist in determining the standard design and 
requirements for a NIRFI system which can be used with populations for whom motion 
restriction is not possible.
The use of motion extraction techniques to improve localization of the NIR signal has been 
demonstrated numerically. Expanding upon this, we aim to examine how the mapping of 
NIRFI data to MNI space might be improved and enhanced through correlation with fMRI 
data. Such a study is important for addressing the question of whether NIRFI can be 
localized to the correct gyri, as previous studies have shown that localization to the 
expected functional area can be problematic due to artifact errors. This is significant in 
critically assessing the efficacy and validity of using NIRFI techniques in currently non-
imageable populations.
2
Study Objectives 
Primary Objectives: Our first primary objective is to collect functional optical imaging 
data from healthy volunteers, study patterns of near-surface cortical activation during 
performance of known tasks, and validate results with published literature. Our second 

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 6 of 28
primary objective is to validate NIRFI imaging data results with fMRI data and assess the 
uses of NIRS techniques as a developing neuroimaging modality.
Secondary Objectives: Our secondary objective is to assess neurovascular changes, 
including blood volume and blood oxygenation fluctuations, during functional events and 
validate observed changes with published literature. In addition, we will assess any 
significant issues associated with NIRS optode placement and evaluate the effect of subject 
motion on data collection, quality, and noise.
3
Subjects
a. Description of study populations 
We will recruit a sample of 250 healthy adults.
b. Inclusion criteria
Age 18 years or greater.
c. Exclusion criteria

Healthy volunteers with any skin disease.

Healthy volunteers with any past or present vascular disease.

Known adverse reaction to latex.

Any medical condition that, in the opinion of the Principal Investigator would 
preclude the inclusion of a patient onto this research study.

Unable or unwilling to give informed consent.
4
Study Design and Methods
a.
Study overview 
This study will be conducted in a group of Healthy Volunteers, all of which will 
present at two outpatient visits (1 visit for screening, consenting and general 
health check and 2 visit for experimental imaging measurement). Each healthy 
volunteer will be admitted for screening, consenting and general health check and 
imaging studies related to functional brain activation. The subject will go through 
neuropsychological tests first, and then will perform the selected protocol for 
functional imaging studies. 
The qualifications of each healthy volunteer will be discussed with the Principal 
Investigator, Study Chairperson, or an Associate Investigator involved with the 
medical conduct of the study. Once it has been determined that a healthy 
volunteer qualifies, has read the informed consent, has had an opportunity to 
discuss the protocol with Principal Investigator, Study Chairperson, or an 
Associate Investigator involved with the medical conduct of the study, and wishes 
to participate, informed consent will be obtained from the healthy volunteer, as 
documented by a signed statement of informed consent approved by the IRB.

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 7 of 28
This protocol is not linked to any other active or prospective protocols at the NIH, 
and study participants are not required to participate in any other protocols.
b. Recruitment
Adult participants will be recruited through the NIH Healthy Volunteer office. All 
recruitment materials and venues will be approved by the IRB. 
c.
Screening, consenting and general health check 
After expressing interest in participating in the study via e-mail or phone call, a 
member of the research staff will contact the subject to verify their identity. 
Questions will be asked to rule potential participants in or out of participation in 
the study, with respect to the basic inclusion and exclusion criteria. 
Prior to any research activities, participants will provide written, informed consent 
before study visit. After signing the consent, participants will have the first visit 
for general health check including: Medical History, physical exam and vital signs. 
Then, the participants will be asked to come back for another visit to do the 
experiments.
d. Study procedures
Behavioral measures: 
Subjects will be asked to take a series of behavioral questionnaires. The specific 
selection of questionnaires administered will depend on the functional tasks 
performed in the experiment. Subject responses to these tests will assist in 
controlling for between-subject differences in behavioral measures which may be 
relevant to task performance capability, ultimately reducing the confounding 
effect these differences may have in subsequent imaging analyses. Participants 
are free to refuse to answer any of the questions contained on any of the 
questionnaires they are given, which has been made clear on the informed consent 
document. These series of behavioral tasks/ questionnaires may include the 
following scales:

Wechsler Adult Intelligence Scale (WAIS) III or IV
WAIS yields three IQ scores (verbal IQ (VIQ), performance IQ, (PIQ) and full 
scale IQ (FSIQ)) and four Index scores (verbal comprehension, perceptual 
organization, working memory and processing speed). Verbal comprehension 
index (VCI) measures general verbal skills (verbal fluency, ability to 
understand and use verbal reasoning, and verbal knowledge). Perceptual 
organization index (PRI) measures non-verbal and in-the-moment reasoning 
skills (ability to examine a problem, draws upon visual-motor and visual-
spatial skills, organizes thoughts, create solutions and then test them). Working 
memory index (WMI) assesses the ability to memorize new information, hold 
it in short-term memory, concentrate, and manipulate that information to 
produce some result or reasoning processes. Processing speed index (PSI) 
assesses skills on focusing on attention and quickly scanning, discriminating 

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 8 of 28
between and sequentially ordering visual information. It refers to the speed at 
which cognitive processes can be carried out. Time varies depending on which 
tests will be administered. 

Kaufman Brief Intelligence Test, Second Edition (Kaufman & Kaufman, 
2004) 
The Kaufman Brief Intelligence Test a short norms-referenced intelligence test 
assessing verbal/crystallized and nonverbal/fluid intelligence. It is designed as 
an intelligence screener for individuals aged 4 to 90 years. Administration take 
approximately 20 minutes to complete the 3 subtests: verbal knowledge, 
riddles, and matrices. This test may be used as an alternative to the WAIS as a 
measure of intelligence.

Symptom Checklist 90-Revised
The Symptom Checklist-90-Revised instrument helps evaluate a broad range 
of psychological problems and symptoms of psychopathology.

Wisconsin Card Sort Test
The Wisconsin Card Sort Test measures neuropsychological functioning, 
assessing abstract thinking, cognitive flexibility and executive function. 

PhenX Broad Psychopathology Screen (Appendix A)
The PhenX Broad Psychopathology screen provides basic information 
regarding participants’ psychological functioning over the past two weeks. It 
is part of the PhenX Toolkit, a toolkit funded by the National Human Genome 
Research Institute (NHGRI) that consists of measures developed to standardize 
phenotypic 
measurement 
in 
biomedical 
research 
studies 
(https://www.phenx.org/). It is commonly used in research to ensure that 
participants have no current psychological symptoms (i.e., within the past two 
weeks). All participants who participate in this protocol are recruited through 
the healthy volunteer office at the NIH; however, this screen has been added 
to ensure that the investigators are as thorough as possible in controlling for 
any potential between-subject variance in our sample during data analysis.

Health History Questionnaire (Appendix B)
The health history questionnaire was created to provide a broad overview of 
participants’ past and current health issues that could interfere with their 
imaging measurements or introduce variance into the data at their time of 
participation. This information will be used to control for any health 
differences in our participants’ by using any relevant health variables as 
covariates in our analyses. Again, all participants who participate in this 
protocol are recruited through the healthy volunteer office at the NIH; 
however, this questionnaire has been added to ensure that the investigators are 
as thorough as possible in controlling for any potential between-subject 
variance in our sample during data analysis.

The Autism Spectrum Quotient (Appendix C)

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 9 of 28
The Autism Spectrum Quotient (AQ) is a validated, self-report questionnaire 
that provides a continuous measure of social functioning (Baron-Cohen et al., 
2001). This scale has been shown to reflect a continuum of social functioning 
ability in typically developing controls, as well as clinical populations. Since 
some of the imaging tasks involve social scenarios, such as the self-reflection 
task (see Section 3.2.2.3) it is important to take all participants’ level of social 
functioning ability into account. Despite its name, this scale is in no way a 
diagnostic tool and is only intended for research purposes. It will only serve as 
a continuous measure of social functioning to be controlled for in subsequent 
analyses, as has been shown to be important in many previous studies of 
typically developing populations (Ruzich et al., 2015; Nummenmaa et al., 
2012; Hasegawa et al., 2013; con dem Hagen et al., 2010).

The Cognitive Flexibility Inventory (Appendix D)
The Cognitive Flexibility Inventory (CFI) is a validated self-report 
questionnaire that measures participant’s day-to-day ability to adapt to changes 
in their environment and cognitive stressors (Dennis et al., 2010). The scores 
on this scale directly pertain to participants’ behavioral inhibition ability and 
will be important to factor into analyses considering their performance on 
behavioral inhibition tasks, such as the go/no-go (Section 3.2.2.4).

The Adult Repetitive Behavior Questionnaire, 2nd Edition (Appendix E)
Like the CFI, the Adult Repetitive Behavior Questionnaire, 2nd Edition (RBQ-
2A) is a validated self-report behavioral scale which measures participants’ 
daily repetitive behavior repertoire that are associated with behavioral 
inflexibility (Barrett et al., 2015) and was developed in neurotypical adults. It 
provides a continuous measure of repetitive behavior, which again is important 
information to consider during analyses involving tasks pertaining to 
behavioral flexibility, such as the Go/No-Go Task (Section 3.2.2.4)

The Edinburgh Handedness Inventory (Appendix F)
The Edinburgh Handedness Inventory (EDI; Old Field, 1971) is a commonly 
used, validated self-report measure of participants’ handedness as assessed by 
which hand their use to perform everyday activities. It provides a score which 
indicated participants’ degree of left or right handedness. It is important to take 
this into account in imaging analyses due to laterality differences in neural 
structure related to handedness (Willems et al., 2014).

Psychopathic Personality Inventory -Revised (PPI-R) (Appendix G)
The PPI-R score is a self report questionnaire to measure emotional 
callousness (coldheartedness), carefree non-planfulness, social influence and 
some other traits, which form global psychopathy traits, in adults (Lilienfeld, 
Widows et al. 2005). PPI-R has been designed to assess those traits in healthy 
people without considering any links to anti-social or criminal behaviors. It 
also can find a pattern in the responses to control for careless or random 
responses. Since some of the imaging studies on human decision making 
involve different factors such as social influences as well as personal traits it 
is important to take all participants’ level of social and personal functioning 
ability into account. Despite its name, this score is in no way a diagnostic tool 
and is only intended for research purposes. It will only serve as a continuous 

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 10 of 28
measure of social and personal functioning to be controlled for in subsequent 
analyses, and has been shown to be important in many previous studies of 
typically developing populations (Rilling, Glenn et al. 2007, Harenski, Kim et 
al. 2009).

Visual Aural Read/Write Kinesthetic (Appendix H)
VARK is a self-assessment learning style (© Copyright Version 7.8 (2014) 
held by VARK Learn Limited, Christchurch, New Zealand). The results of this 
test indicate preferences rather than strengths and can be used for assessing 
learning styles. This questionnaire consists of 16 multiple-choice questions 
regarding subjects’ preferences when conducting a variety of activities. For 
instance, subjects will choose how they prefer to learn something for the first 
time or communicate with others. The multiple choice would allow subjects to 
select the preferred methods to deal with the given situation. The VARK 
questionnaire and the results focus on how people like to receive information 
or deliver their communication. We use the VARK score to find the correlation 
between learning preferences and brain activation during performance of the 
cognitive tasks that use different stimuli such as auditory and visual.
Biological Measures:
fNIRS Devices & Application: After completing these questionnaires, subjects 
will be seated on an adjustable chair. An fNIR device will be applied to the 
appropriate head position on the head and held in place by an adjustable band. One 
of two fNIRS devices will be used to assess participants in this protocol. The 
optode array in Device 1 (fNIRSoft) is most commonly used for assessment of 
hemodynamic activity in the prefrontal region, while Device 2 (NIRScout) can be 
applied to measure activity from other areas of the cortex. Due to this specificity, 
both devices have been included in the protocol. Selection of the device used will 
depend on the aims of the experiment and nature of the tasks performed by the 
subject.
Device 1 – fNIRSoft: This NIRFI device consists of three rows of sources 
and detectors, organized into a band. The bottom and top row of this band 
each contain 5 detectors, while the middle row contains 4 sources. The 
system is designed to cover the forehead of the subject and capture 
activations that occur in the prefrontal cortex region. The current version of 
this device being used in the protocol is the fNIR Devices 2000S imager 
(fNIR Devices, LLC). Specifically, This device uses light emitting diodes 
(LEDs) provide highly monochromatic sources with very fast time 
sequencing (50 msec), available at 730, 805, and 850 nm. These three LEDs 
are contained in a 3 mm diameter housing and are operated as central 
clustered sources with circumferential detectors. A power consumption of 
approximately 0.2 watt, time shared at 1 msec is typical. The light intensities 
are within FDA requirements for Class I devices (<100 mW) for eye safety, 
and do not require further FDA approval. Published results from tests 
utilizing this device were performed at Drexel University, where they were 
also controlled by an assigned safety engineer and reviewed by the Office 

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 11 of 28
of Regulatory Research Compliance. This device has already been approved 
for human subject studies by the Drexel University Institutional Review 
Board. 
Device 2 - NIRScout: The NIRx NIRScout 8x12 fNIRS imaging system is 
a second NIRFI device which may also be used to measure cerebral 
hemodynamic activity. This device includes 8 laser sources (Class 1 lasers 
operating at 760nm and 850nm) which shine low-intensity levels of infrared 
light onto the participant’s scalp, and 12 detectors (Avalanche photodiode). 
The device also includes 8 additional small detectors of the same type used 
to create short source-detector separation channels. The sources and 
detectors will be fit into a comfortable elastic cap worn by the participant. 
Like the fNIR Devices system, this is a safe, passive, and noninvasive 
method of measuring the oxygenation content of cortical blood flow.
Device 3 – Brain Products actiCAP
In conjunction with NIRFI, electroencephalogram (EEG) measurement of 
cortical neural activity is important for the validation of NIRS as well as 
enhancing the temporal resolution of measured activity. The NIRx 
NIRScout system, previously discussed in detail, is designed to operate in 
an integrated fashion with EEG systems produced by Brain Products LLC.  
Specifically, the Brain Products actiCAP is a 32-channel silver chloride 
electrode array which can be worn in combination with a variable number 
of NIRS source-detector pairs integrated at key anatomical sites. Like both 
the fNIR Devices and NIRScout devices, this is a safe, passive, and 
noninvasive method of measuring cortical brain activity which is commonly 
used for the functional assessment of coordinated neural activity.
Physiological measures: Surface electrodes and/or a respiration monitor will 
also be applied to subjects to monitor various physiological signals, such are 
heart rate, respiration, skin conductance response, and blood flow. These signals 
are commonly collected in conjunction with neuroimaging data, particularly 
fMRI studies, as they can then be used to reduce noise in the neuroimaging 
signal due to physiological interference (Caballero-Gaudes & Reynolds, 2017). 
Physiological signals such as heart rate, respiration, skin conductance response 
will be collected through the BioPac MP160, a physiological data acquisition and 
analysis system. The BioPac MP160 can measure several physiological signals 
such as electrocardiogram (ECG), respiration rate, skin conductance response, 
through the use of surface electrodes and signal transducers (e.g., a respiratory 
effort transducer). The BioPac MP160 is a commercially available product from 
BioPac Systems, Inc (Goleta, CA). Electrodes and transducers will be applied to 
each subject to monitor physiological signals throughout their data collection 
session so that they can later be compared to their neuroimaging (fNIRS) data. 
Blood flow measurement: Blood flow will be measured using a Laser speckle 
imaging (LSI) device. LSI is a NIRS technology that is sensitive to flow. LSI 

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 12 of 28
consists of a coherent non-collimated light source (typically 785 nm) and a camera 
that images the illuminated tissue area. When biological tissue is illuminated with 
coherent light, an interference pattern will be formed at the detector, known as a 
speckle pattern. LSI is based on the dynamic change in this backscattered light as 
a result of interaction with red blood cells. Blood flow speed affects the pattern 
captured by the camera. Characterizing these changes in the pattern allows for a 
measurement of blood flow in the skin microvasculature as well as cardiac 
parameters. For this experiment, we are going to use a figure tip LSI, which was 
built in the Section on Biomedical Optics, NICHD. This device consists of a light 
source on one side of the finger or toe, and a small CMOS camera with a pinhole 
aperture making direct contact with the other side of the finger or toe. Blood flow 
and cardiac parameters measured by LSI will be used to remove systemic 
physiological noise from cerebral hemodynamic responses (fNIRS). Like other 
fNIRS techniques, LSI uses low-intensity levels of infrared light, which is safe, 
passive, and noninvasive. 
Eye tracking: 
While completing select cognitive tasks, including the Anti-saccade, emotional 
Anti-saccade, and Flanker tasks, the Eyegaze Edge eyetracker system (LC 
Technologies) will be used to monitor participant gaze and eye movement while 
looking at stimuli presented on a computer screen. This is necessary for the 
assessment of saccadic eye movements, which are rapid, simultaneous movement 
of both eyes in the same direction occurring between two or more phases of visual 
fixation, as well as tracking smooth pursuit eye movement and attention.
The Eyegaze Edge system utilizes a binocular, double fixed-camera system to 
observe the movement of both eyes at a sampling rate of 120 Hertz. Video cameras 
are positioned below a computer monitor, where a small, low power, infrared light 
emitting diode (LED) located at the center of the camera lens illuminates the eye. 
Gaze detection is measured using the pupil-center corneal reflection (PCCR) 
method, a noninvasive technique in which near-infrared light is shone towards the 
participant’s pupils, causing reflections in both the pupil and the cornea. The 
vector between the corneal reflection and the pupil center are detected and tracked 
by an infrared camera. The use of both infrared light sources and detectors is 
critical for the accuracy of detecting corneal reflection, as infrared light provides 
much higher levels of contrast. Importantly, LED illumination of the eyes is safe, 
as the illumination is approximately 20 percent of the HEW maximum permissible 
exposure at a range of 15 inches (LaMarre, 1977).
The Eyegaze Edge system and PCCR method is a well-used, standard eye tracking 
method which is passive, noninvasive, and has a negligible effect on the user. 
During eye tracking, participants will be seated in a comfortable position with the 
stimulus presentation screen fixed at eye level. The Eyegaze Edge system works 
with both glasses and contact lenses, as its automatic calibration procedure 
accommodates the reflective properties of lenses. It also tolerant of minor head 

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 13 of 28
movements and accommodates a wide range of variation in the eye image, 
including shape, brightness, and other features.
Tasks: We propose to conduct a wide battery of experiments using NIRFI to assess 
our ability to map functional events in the near-surface of the cortex. Each task 
will be based on no-risk models from existing fMRI literature for comparative 
purposes. Here we provide references of various example studies using executive 
activations. We will study CBF in the prefrontal cortex while the subject will judge 
event complexity task (Krueger et al., 2009), judge emotional stimulus related to 
the self (Johnson et al., 2002), and perform multitask (Koechlin et al, 1999), as 
well as a variety of cognitive tasks based on those previously used in the fMRI 
literature 
The task details are outlined below:
Event Complexity task
Subjects will be asked to make a dichotomous decision regarding the complexity 
of daily life activities. In particular, participants are asked to rate each activity in 
terms of the number of events involved in the activity. Each activity may consist 
of few events (e.g. ‘stirring a cup of coffee’) or may consist of many events (e.g. 
‘planning a wedding’).
Multi-task
Subjects will perform a letter-matching task in four conditions including a control, 
a delay, a dual-task and a branching condition. The subject will respond to visually 
presented letter (500ms duration) by pressing response buttons with their right or 
left hand. 
Self Reflection
Subjects will be asked to make decision about themselves on specific statement 
requiring self-evaluation in the domains of mood, social interactions, cognitive 
and physical abilities. In the control condition, participants will make decisions 
about statements of factual knowledge.
Go/No-Go Task
Subjects will be asked to complete go/no-go tasks on the computer. The go/no-go 
task is a commonly used cognitive task to assess behavioral inhibition abilities in 
neuroimaging studies. During the task, subjects will respond to visual stimuli 
presented on a computer by either pressing a button or inhibiting the button press 
response.
Cerebral Autoregulation Task: 

Sit-to-Stand Procedure
Subjects will undergo a passive orthostatic test to quantify cerebral 
autoregulation through an active sit-to-stand procedure. Cerebral autoregulation 
and cerebral blood perfusion have been studied through fMRI using arterial spin 

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 14 of 28
labeling; however, conducting such a measurement through fNIRS is less time 
consuming and more economical. Monitoring of fNIRS and physiological signals 
will be performed throughout, as previously described. In the sit-to-stand 
procedure, subjects will stand in an upright posture for a short, standardized time 
period (< 1 minute). They will then be instructed to transition from the standing 
to seated position, where they will hold an upright seated posture. This task will 
be repeated for multiple trials, with a short rest period between each trial (Lee at 
al., 2014). This is an active although noninvasive and nonstrenuous procedure 
which is both a safe and standard method for the evaluation of orthostatic 
tolerance. Moreover, physiological measures (such as heart rate) will be recorded 
throughout the procedure to monitor exertion levels.
A limitation of a sit-to-stand task is that measured cerebral activities using fNIRS 
and EEG could be contaminated by movement artifacts associated with the 
procedure because this experiment used wire-devices/equipment. An alternative 
cerebral autoregulation task which does not require body movement is a cold 
pressor test. 

Cold pressor test
Cold pressor test (CPT) is a long-standing, simple, and validated physiological 
tool to evoke temporary increases in the arterial blood pressure with an elevation 
in the sympathetic nerve activity (an autonomic neural control of the cerebral 
circulation) (Hines et al., 1936; Wood et al. 1984; Menkes et al. 1989; Wirch et al. 
2006; Mourot et sl. 2009; Washio et al., 2020). During this test, in which the 
subject immerses one hand or foot into an ice bath for a short time (e.g.1-3 
minutes), a vascular sympathetic response occurs that results in an increase in 
blood pressure. Due to its effects on blood pressure, CPT has been used 
frequently for predicting hypertension (Wood et al. 1984) and simulating cerebral 
vascular change (Washio et al. 2020). Previous studies have indicated a change in 
cerebrovascular response using fMRI (Hendriks-Balk et al. 2020) and neural 
activities using EEG (Chang et al. 2002). In this protocol, we will monitor 
hemodynamic responses using fNIRS and neural activities using EEG during a 
CPT, in which participants will put their limb in an ice bath (temperature ~ 33.8oF 
to 41oF) for as long as they can with a maximum time of 3 minutes. Temperature 
of the cold bath will be monitored with a standard thermometer. Participant will 
be allowed to remove their limb at any time and the test will be ended to prevent 
severe pain. Physiological signals will be monitored throughout the test to ensure 
a safe condition. Though it is not anticipated to happen, but in case any 
physiological parameter changes to a critical level, the test will be ended 
immediately. After the test, we will correlate our measured signals with literature 
fMRI and EEG data obtained during the task. If a high correlation is found, fNIRS 
can then be used to replace a high-cost fMRI system. 
Computerized Cognitive Tasks:
Subjects will be asked to complete one or more cognitive tasks, similar to the type 
that appear in the NIH Toolbox Cognition Battery. Tasks will be created and 
administered using EPrime Psychology Software Tools and will be based on 

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 15 of 28
established tasks that test various aspects of cognition and executive function (e.g., 
the Flanker test, the Dimensional Change Card Sort test, etc). All of the tasks are 
administered on a computer and consist of the subject being given a set of task 
instructions, then viewing a series of stimuli (consisting of different words, 
images, symbols, numbers, and/or sounds) and responding to these stimuli based 
on the task instructions through a variety of potential behavioral responses (such 
as a button press, mouse click, eye gaze, etc). Computerized cognitive tests such 
as these are common in the fMRI literature and thus comparing fNIRS data. 
Elaborated examples of two potential tasks are described below.
Antisaccade Task
The antisaccade task, commonly used as an assessment of frontal lobe functioning, 
tests a subject’s inhibition of reflexive eye movement towards a presented 
stimulus. Subjects will be asked to fixate their gaze on a non-moving target, such 
as a small dot or cross, presented on a computer screen. A stimulus will be rapidly 
presented without warning in the periphery of the screen, on varying sides of the 
fixation point. The subject will be instructed to make a saccade (a rapid shift in 
gaze) in the direction away from the stimulus.
Flanker Task 
Subjects will be asked to complete a series of Eriksen Flanker Tasks. During the 
task, the subject will be presented with a target stimulus surrounded by congruent, 
incongruent, or neutral flankers. Subjects will be instructed to make directional 
responses associated with the target stimulus by pressing right or left arrow buttons 
on a keyboard. In the congruent condition, flankers elicit the same response as the 
target stimulus. In the incongruent condition, flankers elicit the opposite response 
to the target stimulus. In the neutral condition, flankers have no relationship to the 
directional response. 
Finger tapping task
The finger tapping task is a neuropsychological task which assesses motor 
functioning. Subjects will be asked to perform real, imagined, and imitated finger 
tapping. All finger tapping tasks will be done on the right hand (right-handed 
subject preference). The subject will look at a screen which will display prompts 
to physically tap or imagine tapping their finger. During the imitated finger 
tapping, the subject will watch a video or live demonstration of another person 
tapping his or her finger and be instructed to imitate the same action.
e.
End of participation
After completion of the task, a summary of the task and main purpose of the 
experiment will be given to participants. 
5
Management of Data and Samples
a.
Storage

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 16 of 28
Electronic data records will be stored in a secured database located at NIH and 
paper records will be stored in a locked office in a locked filing cabinet. The data 
will only be accessible to investigators listed on the study protocol. At the 
completion of the study, all data will be stored by the Principal Investigator for 3 
years
b.
Data and sample sharing plan
Data may also be shared with collaborating laboratories at NIH or outside of NIH 
and/or submitted to NIH-designated databases if consent for sharing was 
obtained. Repositories receiving data from this protocol may be open-access or 
restricted access. 
Data will be stripped of identifiers and may be coded (“de-identified”) or unlinked 
from an identifying code (“anonymized”). When coded data is shared, the key to 
the code will not be provided to collaborators but will remain at NIH. Data may 
be shared with investigators and institutions with an FWA or operating under the 
Declaration of Helsinki (DoH) and reported at the time of continuing review. 
Sharing with investigators without an FWA or not operating under the DoH will 
be submitted for prospective IRB approval. Submissions to NIH-sponsored or 
supported databases will be reported at the time of Continuing Review. 
Submission to non-NIH sponsored or supported databases will be submitted for 
prospective IRB approval. 
Required approvals from the collaborating institution will be obtained and 
materials will be shipped in accordance with NIH and federal regulations.
6
Additional Considerations
a. Research with investigational drugs or devices
The system being used to collect fNIRS data is either the fNIR Devices 2000 or the 
NIRx NIRScout Imager. Laser speckle imaging (LSI) device. LSI is a NIRS 
technology and like other fNIRS techniques, LSI uses low-intensity levels of 
infrared light, which is safe, passive, and noninvasive.  ActiCap for the 
measurement of cortical electrical activity will be used conjunction with NIRS 
measurement. ActiCAP is a 32-channel electrode array designed to be worn in 
combination the NIRx NIRFI devices. This is a safe, passive, and noninvasive 
method of measuring cortical brain activity which is commonly used for the 
functional assessment of coordinated neural activity. All of these are commercially 
available, non-invasive devices that are being used for their intended purpose (i.e, 
to measure neural activity). For this reason, the devices are not investigational and 
the study is IDE exempt. 
b. Gene therapy
Not Applicable 

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 17 of 28
7
Risks and Discomforts
There are no known risks associated with the procedures described above, which are widely 
used in cognitive neuroscience studies. Members of the research team will ensure that 
participants remain comfortable throughout the course of the experiment. 
The cold pressor test may provoke pain and discomfort at the limb, which is emerged in 
the cold bath. However, we will emphasize to the participants that their participation is 
completely voluntary and that they can remove their limb from the cold bath at any time 
and we will end the test immediately.
Optical imaging – NIRS, LSI: There are no known risks associated with the procedures 
described above, which are extensively used in different biomedical optic fields. Near-
Infrared Spectroscopy (NIRS) is a noninvasive technique used widely in cognitive 
neuroscience studies.
The silicon cushioning material is used to cover all electronic parts as well as to provide 
comfort to subjects. However, it is conceivable (although never observed during 
experiment to date) that individuals with latex allergies or other abnormal skin conditions 
may experience adverse reactions. This will be made known at every major point of contact 
between the subject and the investigators up to and including the time of informed consent.
The silver-silver chloride (Ag-AgCl) disposable surface electrodes used to record 
physiological signals are peel-and-stick, applied through an adhesive backing, like a band-
aid. There may be mild redness due to removal of these surface electrodes at the end of the 
data collection session, similar to what one would expect when removing a band-aid. The 
respiratory effort transducer is an elastic band worn over the clothes, across the chest that 
is tightened to a snug fit. The snug fit may be mildly irritating to some subjects. If they do 
not tolerate either the electrodes of the respiration band, they are free to refuse to wear 
either or both at any point in the data collection session.
To minimize discomfort, the experimenter will ask subjects about their comfort and adjust 
the procedure as needed.
8
Subject Safety Monitoring
All subjects will be monitored during their data collection session by a member of the 
research team. All study personnel who are in contact with subjects will be First Aid and 
CPR-certified. In the case of a medical emergency, emergency services will be 
immediately contacted, and the participant will be attended to by one of the study personnel 
for proper acute management until emergency services arrive on scene. Any adverse events 
that may occur during the study procedures will be monitored by the Principal Investigator. 
While any risks incurred by participating in the study are minimal, any discomfort while 
wearing the fNIRS cap will be monitored during subjects’ data collection sessions. Since 
the research being conducted is behavioral research, we will not use any toxicity tables for 
safety monitoring. Participation in the present study is voluntary. Healthy adult volunteer 
subjects are free to withdraw from the study at any time without penalty. 

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 18 of 28
9
Outcome Measures
The main outcome measure of this study will be the neuroimaging measure. Specifically, 
graded changes in blood flow and oxygen, measured with NIRS, in response to different 
functional tasks.
10 Statistical Analysis
a.
Analysis of data/ study outcomes
The collated data for each protocol will be compared to the published results from 
pre-existing fMRI studies for both inter and intra patient studies. This will be 
done by comparing the Atlased locations of FNIR data to the Atlased locations 
of the equivalent fMRI study in the literature. We will determine the mean 
coordinates of activation along with the deviation in the population in the same 
manner as the original studies. The populations can then be tested using t-tests 
and ANOVA tests to determine if the population data are from the same or 
different populations.
11 Human Subjects Protection
a.
Subject selection
No selection will be based on race, ethnicity, or gender. Exclusion for healthy 
volunteers under 18 years is based on several issues related to the design of the 
current subject interface. This interface was designed for use on subjects with a 
fully adult sized head. Whilst it is conceivable that a smaller subject would be 
viable it introduces potential confounding factors into the data analysis. 
b.
Justification for inclusion of children 
No children will be included in this study.
c.
Justification for inclusion of other vulnerable subjects 
This study allows the participation of NIH employees, who are considered a 
vulnerable population as per policy 404: Research Involving NIH Staff as 
Subjects. It is a minimal risk study and there are no known risks associated with 
the use of fNIRS with this population. No other vulnerable subjects will be 
included. 
d.
Justification for sensitive procedures 
Not Applicable
e.
Safeguards for NIH Staff or family members of study team members
NIH staff and family members of study team members may be enrolled in this 
study as this population meets the study entry criteria.  Neither participation nor 
refusal to participate as a subject in the research will have an effect, either 
beneficial or adverse, on the participant’s employment or position at NIH.
Every effort will be made to protect participant information, but such 
information may be available in medical records and may be available to 

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 19 of 28
authorized users outside of the study team in both an identifiable an 
unidentifiable manner.
The NIH Frequently Asked Questions (FAQs) for Staff Who are Considering 
Participation in NIH Research will be made available.  Please see section 13 for 
consent of NIH Staff
12 Anticipated Benefit
The potential benefit is that the data collected will assist us in developing a functional brain 
imaging system which will be useable on a much broader patient base than fMRI.
13 Consent Documents and Process
a.
Designation of those obtaining consent
Study investigators designated as able to obtain consent will obtain informed 
consent from healthy adult volunteers.
b.
Consent procedures 
Consent will be obtained for all adult participants. Participants will have an 
opportunity to have their questions answered before being asked to sign the 
consent form. 
c.
Consent documents
The consent form contains all the elements required. The consent forms that have 
been included are as follows: a healthy adult volunteer consent form.
d.
Considerations for Consent of NIH staff, or family members of study team 
members
Consent for NIH staff will be obtained as detailed above with following additional 
protections:
Consent from staff members will be obtained by an individual independent of the 
staff member’s team whenever possible.  Otherwise, the consent procedure will 
be independently monitored by the CC Department of Bioethics Consultation 
Service in order to minimize the risk of undue pressure on the staff member.

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 20 of 28
14 Data and Safety Monitoring
Data and safety monitor
Because this study involves minimal risk to participants, data and safety will be 
monitored by the Principal Investigator, who will be responsible for ensuring validity 
of data collection methods as well as adherence to the protocol.
IRB and DSMC Documentation 

All IRB documentation can be found in iRIS. Principal Investigator, is responsible for 
maintaining IRB documentation, including records of all reviews of the study and 
submissions to the IRB.

Clinical Trial Studies (non IND/IDE) will have random audits performed by the 
NICHD 
Office 
of 
the 
Clinical 
Director 
(policy: 
https://science.nichd.nih.gov/confluence/display/ocd/Protocol+Navigation).
Study Completion
Principal Investigator will maintain subjects’ records for at least three years after 
completion of the study. 
Criteria for stopping the study or suspending enrollment or procedures
If enrollment or procedures for the study are suspended for any reason, the Principal 
and Associate Investigators will conduct a review to determine whether the research 
can resume. 
15 Quality Assurance
a.
Quality assurance monitor
Quality assurance will be monitored by the Principal Investigator and Associate 
Investigators, as well as the NICHD Quality Assurance Program, as the study is 
behavioral in nature and does not involve any intervention or drugs.
b.
Quality assurance plan
Oversight of monitoring will be performed by the NICHD Quality Assurance 
(QA) Program. The NICHD Quality Assurance Program performs annual audits 
randomly on 10% of actively accruing NICHD protocols. 
(https://science.nichd.nih.gov/confluence/display/ocd/Protocol+Navigation).
16 NIH Reporting Requirements
16.1 Reportable events definitions
Please refer to definitions provided in Policy 801: Reporting Research Events.
16.2 Expedited reporting
Please refer to the reporting requirements in Policy 801: Reporting Research Events.
16.3 IRB Requirements for PI Reporting at Continuing Review
Please refer to the reporting requirements in Policy 801: Reporting Research Events.

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 21 of 28
16.4 Reporting to Clinical Director (CD)
Non-compliance, major deviations, unanticipated problems, new information, and 
suspension or termination of research activities will be reported to CD as soon as possible 
and in writing not more than 7 days after the PI first learns of the event. Death will be 
reported within 24 hours after an investigator becomes aware. 
17 Alternatives to Participation 
Subjects do not receive any treatment in this study, nor do they forego any treatment 
in order to participate in this study. The alternative, therefore, is not to participate.
18 Privacy
All research activities will be conducted in as private a setting as possible. All research 
activities will be conducted by researchers who have been trained in the study 
protocol.
19 Confidentiality
a.
For research data and investigator medical records
All data obtained from the research subjects is considered confidential. The data 
will be coded to protect subject confidentiality; electronic records will be stored 
in a secured database located at NIH and paper records will be stored in a locked 
office in a locked filing cabinet. Only study investigators will have access to data.
b.
For stored samples
Not Applicable 
c.
Special precautions 
Results will be published as group data without identifying characteristics. All 
subjects’ information will be kept private. However, if, during the course of the 
study, there is any concern that a subject is at risk of being harmed, study staff 
will report it to the appropriate authorities, as obligated by law.
20 Conflict of Interest
a.
Distribution of NIH guidelines
NIH guidelines on conflict of interest have been distributed to all investigators.
b. Conflict of interest 
There are no conflicts-of-interest to report.
c.
Role of a commercial company or sponsor 
Not Applicable
21 Technology Transfer
Not Applicable 

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 22 of 28
22 Research and Travel Compensation 
Travel expenses for each participant and an accompanying parent(s) will be 
reimbursed according to standard NIH travel reimbursement policy. Subjects will be 
compensated $30 for the first hour and $20 for every hour thereafter if required.

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 23 of 28
23 References
1. Ayers MD, Lawrence DK, (2015). Near-infrared spectroscopy to assess cerebral 
perfusion during Head-up Tilt-table test in Patients with Syncope. Congenital Heart 
Disease. 10 (4), 333-339.
2. Bozkurt A, Onaral B, (2004). Safety Assessment of Near Infrared Light Emitting 
Diodes for Diffuse Optical Measurements. Biomedical Engineering Online, 3(9).
3. Boas, D.A., Dale, A.M., Franceschini, M. A., (2004), “Diffuse optical imaging of 
brain activation: approaches to optimizing image sensitivity, resolution, and 
accuracy,” NeuroImage 23 (Suppl. 1), S275–S288 (2004).
4. Chang, P. F., Arendt-Nielsen, L., & Chen, A. C. (2002). Dynamic changes and 
spatial correlation of EEG activities during cold pressor test in man. Brain 
research bulletin, 57(5), 667-675.
5. Elwell, C. E., Henty, J.R., Leung, T.S., Austing, T., Meek, J.H., Delpy, D.T., and 
Wyatt,J.S., (2005) “Measurement of CMRO2 in neonates undergoing intensive 
care using near infrared spectroscopy,” Adv. Exp. Med. Biol. 566, 263–268.
6. Erickson, S.J. and Godavarty, A., (2008), “Hand-held based near-infrared optical 
imaging devices: A review,” Med Eng Phys.
7. Fukui, Y., Ajichi, Y., and Okada, E., (2003) “Monte Carlo prediction of near-
infrared light propagation in realistic adult and neonatal head models,” Appl. Opt. 
42, 2881–2887.
8. Gratton, E., Toronov, V., Wolf, U., Wolf, M., and Webb, A., (2005), “Measurement 
of brain activity by near-infrared light,” J. Biomed. Opt. 10, 011008.
9. Gibson, A.P., J and Dehghani, H.(2009), “Diffuse optical imaging,” Philosophical 
Transactions of the Royal Society A: Mathematical, Physical and Engineering 
Sciences, 367, p3055-3072.
10. Hendriks-Balk, M. C., Megdiche, F., Pezzi, L., Reynaud, O., Da Costa, S., Bueti, 
D., ... & Wuerzner, G. (2020). Brainstem Correlates of a Cold Pressor Test 
Measured by Ultra-High Field fMRI. Frontiers in neuroscience, 14, 39.
11. Hillman, E.M., (2007), “Optical brain imaging in vivo: techniques and applications 
from animal to man,” J Biomed Opt, 12(5), p. 051402.
12. Hines, E. A., Jr., and Brown, G. E.:  The Cold Pressor Test for Measuring the 
Reactability of the Blood Pressure: Data Concerning 571 Normal and Hypertensive 
Subjects ,  Am. Heart J. 11: 1-9 ( (Jan.) ) 1936.
13. Hoge, R.D., Franceshini, M.A., Covolan, R.J., Huppert, T., Mandeville, J.B., and 
BOAS, D.A., (2005), “Simultaneous recording of task-induced changes in blood 
oxygenation, volume, and flow using diffuse optical imaging and arterial spin-
labeling MRI,” Neuroimage, 25, 701-7.
14. Huppert, T.J., Diamon, S.G., Franceschini, M.A., Boas, D. A., “HomER: a review 
of time-series analysis methods for near-infrared spectroscopy of the brain,” 
Applied Optics, Vol. 48, P. D280.
15. Huppert, T.J., Hoge, R.D., Diamond, S.G., Francechini, M.A., and Boas, D.A., 
(2006), "A temporal comparison of BOLD, ASL, and NIRS hemodynamic 
responses to motor stimuli in adult humans," Neuroimage, 29, 368-82.
16. Johnson, S, Baxter, L, Wilder, L, Pipe, J, Heiserman, J, Prigatano, G. (2002) 
Neural corrolates of self-reflection. Brain, 125, 1808-1814

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 24 of 28
17. Kleinschmidt, A., Obrig, H., Requardt, M., Merboldt, K. D., Dirnagl, U., Villringer, 
A., and Frahm, J., (1996), “Simultaneous recording of cerebral blood oxygenation 
changes during human brain activation by magnetic resonance imaging and near-
infrared spectroscopy,” J. Cereb. Blood Flow Metab. 16, 817–826.
18. Koechlin, E, Basso, G, Pietrini, P, Panzer, S, Grafman, J. (1999) The role of 
anterior prefontal cortex in human cognition. Nature, 13, 148-151.
19. Krueger, F., Spampinato, M.V., Barbey, A.K., Huey, E. D., Morland, T., and 
Grafman, J., (2009)“The frontopolar cortex mediates event knowledge complexity: 
a parametric functional MRI study,” NeuroReport.
20. Lee, SH., Jang, G., Jin, SH., Park, JH., Lee, YJ., Lee, JM., Kim, SJ., An, J., (2014) 
Cortical activities during a stand to sit movement using fNIRS. 2014 14th 
International Conference on Control, Automation and Systems (ICCAS 2014), 279-
281.
21. Mahoney, PJ., Panerai RB., Deverson ST., Hayes PD., Evans DH., (1999) 
Assessment of the thigh cuff technique for measurement of dynamic cerebral 
autoregulation. Stroke, 31(2), 476-480. 
22. Menkes MS, Matthews KA, Krantz DS, Lundberg U, Mead LA, Qaqish B, Liang 
KY, Thomas CB, Pearson TA. Cardiovascular reactivity to the cold pressor test as 
a predictor of hypertension. Hypertension. 1989 Nov;14(5):524-30. doi: 
10.1161/01.hyp.14.5.524. PMID: 2807514.
23. Mourot L, Bouhaddi M, Regnard J. Effects of the cold pressor test on cardiac 
autonomic control in normal subjects. Physiol Res. 2009;58(1):83-91. doi: 
10.33549/physiolres.931360. Epub 2008 Jan 17. PMID: 18198985.
24. Nagamitsu, S., Nagano, M., Yamashita, Y., Takashima, S., and Matsuishi, T., 
“Prefrontal cerebral blood volume patterns while playing video games–a near-
infrared spectroscopy study,” Brain Dev. 28, 315–321 (2006).
25. Okada, E., and Delpy, D.T., (2003) “Near-infrared light propagation in an adult 
head model. I. Modeling of low-level scattering in the cerebrospinal fluid layer,” 
Appl. Opt. 42, 2906–2914.
26. Roy, C. S. & Sherrington, C. S., On the regulation of the blood-supply of the brain, 
J. Physiol. 1890, 1, p. 85.
27. Sowell,E.R., Thompson, P.M., Leonard, C.M., Welcome, S.E., Kan, E., and Toga, 
A.W., (2004) “Longitudinal mapping of cortical thickness and brain growth in 
normal children,” J. Neurosci. 24, 8223–8231.
28. Strangman, G., Culver, J. P., Thompson, J. H., and Boas, D. A. (2002) “A 
quantitative comparison of simultaneous BOLD fMRI and NIRS recordings during 
functional brain activation,” Neuroimage, 17, 719-31.
29. Villringer, A., and Chance, B., (1997), “Noninvasive optical spectroscopy and 
imaging of human brain function,” Trends in Neuroscience, 20(10), p. 435.
30. Wilcox, T., Bortfeld, H., Woods, R., Wruck, E., and Boas, D. A., “Using near-
infrared spectroscopy to assess neural activation during object processing in 
infants,” J. Biomed. Opt. 10, 11010 (2005).
31. Yodh, A. and Boas, D., (2003) eds., Functional Imaging with Diffusing Light (CRC 
Press)

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 25 of 28
32. Zaramella,P., Freato, F., Amigoni, A., Salvadori, S., Marangoni, P., Suppjei, A., 
Schiavo, B., and Chiandetti,L. “Brain auditory activation measured by near-
infrared spectroscopy (NIRS) in neonates,” Pediatr. Res. 49, 213–219 (2001).
33. Baron-Cohen, S., Wheelwright, S., Skinner, R., Martin, J., and Clubley, E. (2001). 
The Autism Spectrum Quotient (AQ): Evidenec from Asperger Syndrome/High 
Functioning Autism, Males and Females, Scientists and Mathematicians. Journal 
of Autism and Developmental Disorders, 31, 5-17.
34. Ruzich, E., Allison, C., Smith, P., Watson, P., Auyeung, B., Ring, H., & Baron-
Cohen, S. (2015). Measuring autistic traits in the general population: a systematic 
review of the Autism-Spectrum Quotient (AQ) in a nonclinical population sample 
of 6,900 typical adult males and females. Molecular autism, 6(1), 2.
35. Nummenmaa, L., Engell, A. D., Von Dem Hagen, E., Henson, R. N., & Calder, A. 
J. (2012). Autism spectrum traits predict the neural response to eye gaze in typical 
individuals. Neuroimage, 59(4), 3356-3363.
36. Hasegawa, N., Kitamura, H., Murakami, H., Kameyama, S., Sasagawa, M., Egawa, 
J., ... & Someya, T. (2013). Neural activity in the posterior superior temporal region 
during eye contact perception correlates with autistic traits. Neuroscience letters, 
549, 45-50.
37. von dem Hagen, E. A., Nummenmaa, L., Yu, R., Engell, A. D., Ewbank, M. P., & 
Calder, A. J. (2010). Autism spectrum traits in the typical population predict 
structure and function in the posterior superior temporal sulcus. Cerebral cortex, 
21(3), 493-500.
38. Dennis, J. P., & Vander Wal, J. S. (2010). The cognitive flexibility inventory: 
Instrument development and estimates of reliability and validity. Cognitive therapy 
and research, 34(3), 241-253.
39. Barrett, S. L., Uljarević, M., Baker, E. K., Richdale, A. L., Jones, C. R. G., & 
Leekam, S. R. (2015). The adult repetitive behaviours questionnaire-2 (RBQ-2A): 
A self-report measure of restricted and repetitive behaviours. Journal of Autism and 
Developmental Disorders, (doi: 10.1007/s10803-015-2514-6)
40. Oldfield, R. C. (1971). The assessment and analysis of handedness: The 
Edinburgh inventory. Neuropsychologia, 9, 97-113.
41. Willems, R. M., Van der Haegen, L., Fisher, S. E., & Francks, C. (2014). On the 
other hand: including left-handers in cognitive neuroscience and neurogenetics. 
Nature Reviews Neuroscience, 15(3), 193.
42. Caballero-Gaudes, C., & Reynolds, R. C. (2017). Methods for cleaning the BOLD 
fMRI signal. Neuroimage, 154, 128-149.
43. Harenski, C. L., et al. (2009). "Neuroticism and psychopathy predict brain 
activation during moral and nonmoral emotion regulation." Cognitive, Affective, 
& Behavioral Neuroscience 9(1): 1-15.
44. Hoge, R. D., et al. (2005). "Simultaneous recording of task-induced changes in 
blood oxygenation, volume, and flow using diffuse optical imaging and arterial 
spin-labeling MRI." Neuroimage 25(3): 701-707.
45. Huppert, T. J., et al. (2006). "A temporal comparison of BOLD, ASL, and NIRS 
hemodynamic responses to motor stimuli in adult humans." Neuroimage 29(2): 
368-382.

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 26 of 28
46. Kleinschmidt, A., et al. (1996). "Simultaneous recording of cerebral blood 
oxygenation changes during human brain activation by magnetic resonance 
imaging and near-infrared spectroscopy." J Cereb Blood Flow Metab 16(5): 817-
826.
47. Lilienfeld, S. O., et al. (2005). "Psychopathic Personality InventoryTM-Revised." 
Social Influence (SOI) 61(65): 97.
48. Rilling, J. K., et al. (2007). "Neural correlates of social cooperation and non-
cooperation as a function of psychopathy." Biological psychiatry 61(11): 1260-
1271.
49. Strangman, G., et al. (2002). "A quantitative comparison of simultaneous BOLD 
fMRI and NIRS recordings during functional brain activation." Neuroimage 
17(2): 719-731.
50. Washio, T., Watanabe, H., & Ogoh, S. (2020). Dynamic cerebral autoregulation 
in anterior and posterior cerebral circulation during cold pressor test. The Journal 
of Physiological Sciences, 70(1), 1-8.
51. Wirch JL, Wolfe LA, Weissgerber TL, Davies GA. Cold pressor test protocol to 
evaluate cardiac autonomic function. Appl Physiol Nutr Metab. 2006 
Jun;31(3):235-43. doi: 10.1139/h05-018. PMID: 16770350.
52. Wood DL, Sheps SG, Elveback LR, Schirger A. Cold pressor test as a predictor of 
hypertension. Hypertension. 1984 May-Jun;6(3):301-6. doi: 
10.1161/01.hyp.6.3.301. PMID: 6735451.

Abbreviated Title: Non-Invasive Optical Imaging
Version Date: 04/01/2022
page 27 of 28
24 Attachments/ Appendices

PhenX Broad Psychopathology Screen (Appendix A)

Health History Questionnaire (Appendix B)

The Autism Spectrum Quotient (Appendix C)

The Cognitive Flexibility Inventory (Appendix D)

The Adult Repetitive Behavior Questionnaire, 2nd Edition (Appendix E)

The Edinburgh Handedness Inventory (Appendix F)

Psychopathic Personality Inventory -Revised (PPI-R) (Appendix G)

Visual Aural Read/Write Kinesthetic (Appendix H)