A Feasibility Study for the Use of Multispectral Optoacoustic Tomography in the Detection of Solid Tumors and Lymph Nodes (MSOT)
Phase: NA
Conditions: Solid Tumor
Sponsor: University of Oklahoma

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CLINICAL STUDY PROTOCOL 
A Feasibility Study for the use of Multispectral Optoacoustic Tomography in the 
Detection of Solid Tumors and Lymph Nodes (MSOT) 
 
 
Principal Investigator: 
Lacey McNally, PhD 
 
Professor 
 
Stephenson Chair Cancer Imaging 
 
Department of Surgery 
 
University of Oklahoma Health Sciences Center  
 
Stephenson Cancer Center 
 
755 Research Park Rm 471 
 
Oklahoma City, OK 73104 
 
lacey-mcnally@ouhsc.edu 
 
 
 
 
 
 
 
Co-Investigator(s): 
Kathleen Moore, MD 
 
Juan C Claros, MD 
 
William C. Dooley, MD 
 
Ronald Squires, MD  
 
Barish H. Edil, MD, FACS 
 
Ajay Jain, MD 
 
Katherine T. Morris, MD 
 
 
 
Sponsor: 
The University of Oklahoma Health Sciences Center 
Stephenson Cancer Center 
 
800 10th Street 
 
Oklahoma City, OK 73104 
 
IDE Number:                              G180105 
NCT Number:                             NCT04365413 
 
 
 
 

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Table of Contents 
 
1.0 
SCHEMA ..................................................................................................................... 4 
2.0 
Introduction and Background ........................................................................................ 5 
2.1 
Introduction and Background .................................................................................... 5 
2.2 
Clinical Studies......................................................................................................... 6 
3.0 
Objectives .................................................................................................................... 8 
3.1 
Primary Objective(s) ................................................................................................. 8 
3.2 
Secondary Objective(s) ............................................................................................ 8 
4.0 
Patient Selection .......................................................................................................... 9 
4.1 
Inclusion Criteria....................................................................................................... 9 
4.2 
Exclusion Criteria ..................................................................................................... 9 
4.3 
Inclusion of Women and Minorities............................................................................ 9 
5.0 
Subject Registration and Enrollment ........................................................................... 10 
5.1 
Required protocol specific regulatory documents..................................................... 10 
5.2 
Patient Registration and Enrollment ........................................................................ 10 
6.0 
Study Outcomes and Study Measures ........................................................................ 10 
6.1 
Primary Outcome ................................................................................................... 10 
6.2 
Secondary Outcomes ............................................................................................. 10 
7.0 
Study Plan ................................................................................................................. 11 
7.1 
Study-Related Activities .......................................................................................... 11 
Study Procedures ........................................................................................................... 11 
7.2 
Duration of Follow Up ............................................................................................. 15 
7.3 
Criteria for Removal from Study .............................................................................. 15 
8.0 
Adverse Events List and Reporting Requirements....................................................... 16 
8.1 
Adverse Event List for MSOT.................................................................................. 16 
8.2 
Adverse Event Characteristics ................................................................................ 16 
8.3 
Serious Adverse Events (SAE) ............................................................................... 16 
8.4 
Unanticipated Adverse Device Effects Reporting Requirements .............................. 17 
8.5 
Reporting of Unanticipated Problems, Adverse Events or Deviations ....................... 17 
8.6 
Post-imaging Procedural Events for MSOT ............................................................. 18 
9.0 
Data and Safety Monitoring ........................................................................................ 18 
9.1 
DSMC Auditing....................................................................................................... 19 
9.2 
Data Handling and Record Keeping ........................................................................ 19 

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9.2.1 Data Quality Assurance .......................................................................................... 19 
9.2.2 Electronic Database and Case Report Forms.......................................................... 19 
9.2.3 Data Disclosure and Subject Confidentiality ............................................................ 19 
9.2.4 Record Retention ................................................................................................... 20 
9.3 
Study Monitoring .................................................................................................... 20 
10.0 
Statistical Considerations ........................................................................................... 20 
10.1 
Analysis of Primary Objective ................................................................................. 20 
10.2 
Analysis of Secondary Objective ............................................................................. 21 
10.3 
Power and Sample Size ......................................................................................... 22 
10.4 
Estimated Accrual Rate .......................................................................................... 23 
10.5 
Estimated Study Length.......................................................................................... 23 
11.0 
ETHICAL AND REGULATORY CONSIDERATIONS................................................... 23 
11.1 
Ethical Conduct of the Study................................................................................... 23 
11.2 
Informed Consent ................................................................................................... 23 
References............................................................................................................................ 25 
Appendix A – Imaging and Tissue Data Collection Form ........................................................ 26 
Appendix B – Image Analysis Form ....................................................................................... 30 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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1.0 
SCHEMA 
 
 
Consent and pre-study data collection 
Measure skin temperature 
MSOT imaging 
Study follow-up 
Measure skin temperature 

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2.0 
Introduction and Background 
2.1 
Introduction and Background 
The accurate detection and localization of cancers in vivo are critical to medical decisions 
and improved treatments. Unfortunately, limitations of contrast (reporter) agents, 
resolution, and restrictions of depth reduce the ability of most imaging methods to detect 
and localize multiple contrast agents simultaneously. Alternatively, optical-fluorescent 
imaging can simultaneously detect multiple biomarkers using fluorescently labeled 
peptides or antibodies as contrast agents; however, these approaches are restricted to 
superficial detection, as light scattering degrades the spatial resolution at increased 
penetration depths greater than 8 mm. Multispectral optoacoustic tomography (MSOT) is 
emerging as an alternative modality that is not restricted by many of the limitations of the 
imaging used in diagnosis and treatment of diseases1. MSOT was initially developed for 
research, but has been adapted for clinical uses2-4 (see Table 1). 
 
Optoacoustic imaging is based upon a “light-in, sound-out,” i.e. shining a light into the 
tissue and when the light hits the tissue, it can send a sound wave which we will capture 
using the MSOT device, approach through which absorption of near-infrared light (NIR) 
within biological tissues generates ultrasonic waves with much less scattering, longer 
range of detection, and higher accuracy compared with traditional optical imaging. The 
optoacoustic approach is unique, with increased optical contrast and signal-to-noise 
ratios1. Optoacoustic imaging retains the advantages of optical imaging, including high 
specificity to identify functional and molecular processes in living organisms with high 
sensitivity. Most tissues are relatively transparent to NIR light in the range of 600 to 900 
nm; therefore, use of NIR light excitation and ultrasound signals renders photon scattering 
irrelevant to image formation, enabling high-resolution images of the biological function 
of tumors and organs. Once the sound waves are generated, they obey the physical laws 
of sound transmission; the intensity of the sound increases with the number of molecules 
excited, but is reduced by distance and the extent of ultrasound diffraction due to different 
densities of tissue. In MSOT, multiple spectral components of NIR light are varied 
automatically to excite specific molecules, permitting accurate tomographic images to be 
constructed from the resulting ultrasonic 
signals. MSOT is also unique in its ability 
to 
detect 
multiple 
contrast 
agents 
simultaneously based upon differential 
spectral shape. 
Changes 
in 
vasculature 
are 
often 
associated with both tumor development 
and 
progression, 
but 
microvascular 
changes occur below the resolution of 
common 
clinical 
imaging 
modalities. 
MSOT identifies 
microvascularity 
and 
tissue 
oxygenation 
by 
hemoglobin 
absorption of multiple wavelengths of light 
to generate high optoacoustic contrast1,4-6. 
Figure 1  Spectra of endogenous 
absorbers that serve as endogenous 
contrast for MSOT 

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Because oxy- and deoxy-hemoglobin each generate a unique optoacoustic signal, both 
oxy- and deoxy-hemoglobin can be observed simultaneously without the addition of 
exogenous contrast agents using MSOT (Figure 1). MSOT can distinguish between 
oxygenation states of hemoglobin, i.e. oxy-hemoglobin vs deoxy-hemoglobin, allowing 
visualization of differential blood saturation by oxygen within tissues, including the 
capability to differentiate between tumor and the surrounding tissue2,7 as well as 
inflammation associated with Crohn’s disease3.  
Imaging of tumors and cancer-related morphologic changes in tissues by MSOT is 
facilitated through exogenous contrast agents, including clinically approved optical dyes 
(e.g., isosulfan blue or indocyanine green)8, markers targeted to cell surface molecules, 
for example, EGFR 11, the tumor microenvironment (e.g., pH)9, and endogenous 
absorbers (i.e., oxyhemoglobin)2,10. Use of MSOT in multiple tissue types and at varying 
depths, i.e., at least 5 cm3,8, can provide functional real-time three-dimensional (3D) 
information at high spatial resolution in vivo1. This ability will have a significant impact on 
clinical care in systemic diseases, including cancer involving multiple organs. The ability 
of MSOT to identify tumors indicates great potential for clinical applicability for solid 
tumors, such as melanoma, head and neck, breast, pancreatic, prostate, colon, and 
potentially, liver cancer. 
 
2.2 
Clinical Studies 
MSOT has been used in research1,6,11,12 and clinical studies2,3,7,8 (including 6 clinical trials 
described in Table 1) and there has been no evidence of serious adverse events 
associated with the use of the device. Using a similar instrument in breast cancer, an 
optoacoustic imaging system from Seno Medical Instruments has received a CE mark in 
April 2014. Confirmatory studies for product approval of this particular device and specific 
indication have been started during the last years (with a focus on safety and 
performance).  Presently, there has not been any association of phototoxicity associated 
with the use of MSOT in any clinical trial (Table 1). 
 
 

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Table 1 
Clinical Trials using MSOT 
Indication 
Purpose of trial 
Patients 
References 
Status 
Peripheral 
athero- 
sclerosis 
– 
healthy 
volunteers 
The current project comprises a proof of principal 
study to test the technical feasibility of the use of 
Multispectral Optoacoustic Tomography (MSOT) 
in 
healthy 
volunteers 
for 
non-invasive 
measurements and imaging of perfusion and 
oxygenation in the lower legs. By measuring 
peripheral blood oxygenation and perfusion with a 
prototype handheld MSOT probe and comparing 
that to standard ultrasound imaging, pulse-oximetry 
and standard ankle-brachial index as a gold 
standard, the diagnostic accuracy and measurement 
ranges in physiologic conditions as determined by 
MSOT will be tested. 
10  
(The 
Netherlands) 
NTR4125 
http://w w w.trialregis
ter.nl/trialreg/admin
/rctview .asp?TC=4
125 
Published 
in 
Radiology 
Limb ischemia, 
athero- 
sclerosis 
This study aims to investigate the feasibility and 
clinical performance of MSOT imaging of the feet 
in Peripheral Vascular (PVD) patients. Our main 
objective is to investigate and obtain new 
information about the state of arteries in PVD 
patients using MSOT. Our second objective is to 
investigate the use of MSOT compared to duplex 
ultrasound 
and 
standard 
diagnostic work-up 
(including Fontaine classification) in providing 
more information on microvascular perfusion and 
oxygenation in the extremities in relation to severity 
of the PVD. The patients will be imaged on both feet 
at particular landmarks, which include large arteries 
(arteria tibialis posterior and dorsalis pedis) and 
microvasculature (on each of the toes). The images 
will be compared to duplex ultrasound images of the 
large arteries, which are acquired as part of the 
standard of care, as well as other available 
diagnostic information, including the Fontaine 
classification. 
24 
(The 
Netherlands) 
NTR4461 
http://w w w.trialregis
ter.nl/trialreg/admin
/rctview .asp?TC=4
461 
Closed; 
Data 
analysis 
ongoing 
Melanoma 
This prospective study tests if photoacoustic 
tomography (PAT) and high-resolution ultrasound 
in combination with an indocyanine green injection 
is the first valid method to exactly assess the sentinel 
node status in melanoma patients in a non- invasive 
way 
and 
without exposing the patient to 
radioactivity. Therefore, preoperative, in vivo PAT 
results will be compared to the in vivo results based 
on standard diagnostics. Additionally, postoperative 
ex vivo PAT results followed by histological 
assessment will be compared. 
40/120 
(Germany) 
DRKS00006329 
http://w w w.drks.de/
drks_w eb/navigate.
do?navigationId=tri
al.HTML&TRIAL_I
D=DRKS00006329 
 
DRKS00005447 
http://w w w.drks.de/
drks_w eb/navigate.
do?navigationId=tri
al.HTML&TRIAL_I
D=DRKS00005447 
First 
20 
patients: 
Published 
in Sci. 
Transl. 
Med. 
http://stm.
sciencema
g.org/cont
ent/7/317/
317ra199 
Next 
60 
patients: 
Data 
analysis 
Crohn’s 
disease/ 
Ulcerative 
colitis 
The proposed study is designed as a pilot study to 
evaluate the usefulness of MSOT for the evaluation 
of disease activity in IBD patients. As current 
methods 
for 
the 
evaluation 
of 
intestinal 
inflammation in IBD have relevant restrictions, 
new, non- invasive, quantitative, and accurate 
modalities are urgently needed. In comparison to 
other techniques, 
MSOT 
provides 
relevant 
advantages such as non-invasive imaging, high 
spatial resolution (this is of special importance for 
the evaluation of the intestine, which can be only a 
344 
(Germany) 
 
 
NCT02622139 
https://clinicaltrials.
gov/ct2/show /NCT0
2622139?term=MS
OT&amp;rank=1 
Recruiting; 
First 
case 
study 
published 
in Gastro- 
enterology 
in motion 

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few millimeters thick), and highly sensitive 
detection of specific molecules such as total 
hemoglobin or oxy- /deoxygenated hemoglobin. 
These advantages suggest MSOT as an outstanding 
technique for evaluating disease activity in IBD. 
Breast Cancer 
Evaluation of breast cancer based upon oxy- and 
deoxy-hemoglobin. MSOT provided high special 
resolution of breast tissue at centimeter depths.  
Germany 
 
Clinical 
Cancer 
Research 
Alopecia 
Dermal Papilla cells (DPCs) of hair follicle units 
(HFU) derived from balding and non-balding 
regions of the scalp have differences in terms of 
their 
genetic, 
morphologic 
and 
functional 
characteristics. 
One of the study goals is to evaluate the ability of a 
new technology called multi-spectral optoacoustic 
tomography (MSOT) to non-invasively image the 
structural information of hairs and surroundings. 
20 
(Singapore) 
Not available 
First 
case 
study 
submitted 
for 
publication 
Basal 
cell 
carcinoma 
The purpose of this research study is to evaluate if 
MSOT 
can 
accurately 
map 
non-melanoma 
carcinomas with the objective to determine lesion 
dimensions to aide in surgical removal. Two- and 
three-dimensional handheld MSOT probes are 
compared in terms of their performance in mapping 
the non-melanoma lesions. 
Not 
available 
(Singapore) 
Not available 
First 
case 
study 
submitted 
for 
publication 
3.0 
Objectives 
This IDE FDA approved study will involve the interaction with patients that are scheduled 
for routine standard of care surgery. It will be a single-arm study that is designed to 
provide safety information regarding the use of the Acuity MSOT device in the clinical 
setting and the ability of MSOT imaging data to correlate with clinical findings identified 
via pathology. We will initially study the safety data in 10 patients, and will expand the 
study with FDA approval to include 40 more patients after an interim analysis (refer to 
section 10.3 for more details).  
3.1 
Primary Objective(s) 
 
3.1.1 To collect safety data on patients in whom MSOT was used to 
image tumor or lymph nodes  
3.1.2 To evaluate skin temperature pre and post imaging (pre- and post-
surgery) as part of the safety evaluation of MSOT 
3.2 
Secondary Objective(s) 
3.2.1 To detect tumor or lymph node by MSOT before and after surgical 
removal  
3.2.2 To determine tumor positivity based on tissue pathology and 
detection of oxy- and deoxy-hemoglobin using MSOT localization  
3.2.3 To determine tumor volume by MSOT before and after surgical 
resection 

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4.0 
Patient Selection 
4.1 
Inclusion Criteria  
4.1.1 Patients with an identified solid tumor, i.e. breast (Stage I-IV), 
melanoma (Stage I-IV), HNSCC (Stage I-III), pancreatic (Stage I-III), 
ovarian (Stage I-IV) that is scheduled for surgical removal of the 
tumor and completed standard imaging prior to surgery  
4.1.2 Have acceptable hematologic status [total hemoglobin (tHb) ≥ 10 
mg/dL]  
4.1.3 Patients ≥ 18 yrs of age  
4.1.4 Patient provided a signed and dated informed consent  
4.1.5 Willing to comply with study procedures and be available for the 
duration of the study 
4.1.6 Ability to understand and the willingness to sign an IRB-approved 
informed consent document.  
4.2 
Exclusion Criteria  
 
4.2.1 Patients with central nervous system tumors  
4.2.2 Patients with a tattoo over the surgical site  
4.2.3 Pregnant women  
4.2.4 Women who are breastfeeding  
4.2.5 Systemic or local infection  
4.2.6 Any systemic anomaly during the pre-op assessment preventing 
patient participation in the study 
4.2.7 Any febrile illness that precludes or delays participation 
preoperatively  
4.2.8 Anything that would put the participant at increased risk or preclude 
compliance with the study 
4.2.9 Patients with Stage IV pancreatic cancer, Stage IV HNSCC are not 
surgical candidates and therefore excluded from this study  
4.3 
Inclusion of Women and Minorities 
• Women and individuals of all races and ethnicities who meet the above-described 
eligibility criteria are eligible for this trial  
• The study consent form will also be provided in Spanish for Spanish-speaking 
participants. Based on population estimates, we expect approximately 52% of 
participants to be women. Translating this to our expanded sample size estimate 
of 50, we plan to enroll at least 26 women. We plan to enroll at least 6% Native 
American, Hispanic/Latino, or African American (N=3). This would result in at least 
6 patients of Fitzpatrick Skin Type I, 40 patients of Fitzpatrick Skin Type II-IV, and 
4 patients of Fitzpatrick Skin Type V-VI.  Because of the population distribution of 
our area we do not expect to accrue many individuals of Asian/Pacific Islander 
ancestry.  

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5.0 
Subject Registration and Enrollment 
5.1 
Required protocol specific regulatory documents 
This protocol, the Informed Consent document, any information to be given to the 
patient, and relevant supporting information must be submitted to the IRB by the 
Principal Investigator and reviewed and approved by the IRB before the study is 
activated. 
5.2 
Patient Registration and Enrollment 
 
Patients must have signed and dated all applicable consents and authorization 
forms to be registered in the Clinical Trials Management System (CTMS) system, 
which is sponsored by The University of Oklahoma Stephenson Cancer Center 
(OU-SCC). 
First, the study site should send the required documentation to SCC-IIT-
Office@ouhsc.edu for confirmation of eligibility. A Screening identification 
(Screening ID) number will be provided by OU-SCC research staff. After all 
screening procedures and assessments have been completed and eligibility has 
been established, the subject Study ID number will be generated by OU-SCC. 
Once the patient has been provided a Study ID number, only the Study ID number 
should be used.  
Patients must not start protocol procedures prior to registration and enrollment. A 
patient will start protocol procedures only after the pre-treatment evaluation is 
complete and eligibility criteria have been met.  
NOTE: Per the Institutional Review Board (IRB) reporting, a patient is considered 
accrued once he or she signs a consent form for the study. A patient is considered 
enrolled once the patient begins treatment.  
6.0 
Study Outcomes and Study Measures 
6.1 
Primary Outcome 
 
6.1.1 Adverse events as characterized by CTCAE v5.0 in patients that may 
result from MSOT imaging (≥44oC)13  
6.1.2 Measurement of skin temperature pre- and post-MSOT imaging (pre- 
and post-surgery 4 measurements total) with a touch thermometer 
as part of the safety evaluation of the MSOT device. The 
thermometer will be placed onto the skin until a temperature appears, 
about 1 minute, and the temperature will be recorded. 
6.2 
Secondary Outcomes 
 

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6.2.1 Tumor positivity as based on standard clinical laboratory techniques 
and signal detection of oxy- and deoxy-hemoglobin using MSOT 
localization  
6.2.2 Tumor volumes as estimated by MSOT and by standard clinical 
laboratory techniques 
6.2.3 Identification of tumor and/or lymph nodes based upon either 
Isosulfan Blue or IC Green dye 
 
7.0 
Study Plan 
7.1 
Study-Related Activities  
Study Procedures 
• Upon receipt of a signed consent form, the 
research study team will generate a unique 
identification number so that subjects are 
de-identified 
for analysis of data. Any 
standard imaging measurements will be 
added in Appendix A and B. This unique 
identification number will be used on the 
case report form (Appendix A and B).  
• Patients will go through standard care 
procedures to prepare for surgery. Medically 
established diagnostic procedures will be 
used to detect tumor and lymph node 
margins.  
• To evaluate the safety of MSOT (Figure 2), 
the device will be used to obtain images of 
the tumor or lymph node margins for investigational use only to compare to clinical 
pathology and patient’s medical record.  
• As part of the standard of care surgery preparation for cancer resection for all tumor 
types, patients will be injected with standard doses of Isosulfan Blue/ Indocyanine (IC) 
Green for either lymph node or tumor identification as part of the standard surgical 
procedure. The MSOT device will detect the presence of dye at the standard dose of 
dye used for surgical evaluation of tissue. 
Figure 2 The MSOT Acuity 

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• Images will be obtained pre- and post-surgery (example simulation of operating room 
shown Figure 3 ). All 
imaging will be done 
in a closed surgical 
patient; 
no imaging 
will be performed on 
an 
open 
surgical 
patient. 
Dr. 
Lacey 
McNally 
has 
been 
trained in the use of 
the MSOT Acuity by 
the manufacturer, and 
she will be the MSOT 
operator for all MSOT 
imaging for this study. 
Dr. McNally will train 
the 
surgeon 
and 
surgical staff in the 
proper procedure for 
using the handheld 
detector and will maintain training records for this purpose. Only Dr. McNally and her 
research staff will perform post-imaging analysis on the de-identified digital images 
stored on the MSOT Acuity. The pathologist and clinician will perform all clinical 
evaluations of tumor samples as well as post-imaging assessment of treatment area 
and report where indicated on the imaging case report form. MSOT imaging will be for 
research only and no treatment decisions will be based on the MSOT images 
obtained. 
• Logistics: The MSOT device will be used in The University of Oklahoma Health 
Sciences Center (OUHSC), and will reside in a temperature controlled and locked 
room. 
Table 2 
Clinical Protocol for MSOT Imaging 
Step 
Description 
1 
Prior to the time of surgery, the MSOT operator will ensure that the MSOT Acuity has 
been cleaned according to the manufacturer’s recommendations found in the Instruction 
Manual -MSOT Acuity Cleaning & Disinfection. 
2 
A case report form with a unique identification number will be generated for each study 
participant (Appendix A). 
3 
The MSOT operator will maneuver the MSOT Acuity to the operating room and plug unit 
into proper electrical outlet. The MSOT Acuity will be positioned to allow easy access to 
the region of interest for imaging. The wheels will be locked into place to prevent 
acquisition unit from rolling during imaging. 
Figure 3: Demonstration of MSOT scanning 
procedure.A. Handheld detector covered with 
drape; B. surgeon; C. MSOT Acuity; D. trained 
MSOT operator. 

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4 
The adjustable arm holding the monitor and keyboard 
will be locked into position for use by the MSOT 
operator for image acquisition. 
5 
The MSOT operator will turn 
the key on the front panel to 
switch on the Acuity system, 
wait until the logon screen 
appears and log in. 
  
 
6 
Following the login, 
ViewMSOT™ automatically 
starts, initializes the 
hardware and opens the 
Select patient dialog. 
 
 
7 
The MSOT operator will create a new patient record using the unique identification 
number as the Patient ID. 
8 
The MSOT operator will Click on the 
Examination tab in the task bar to open 
the Examination screen. 
 
9 
The Select preset* window will open, 
and the MSOT operator will choose the 
Hb, HbOmelanin application to access the 
Examination screen. Check box when 
complete on Imaging Case Report 
form. 
*NOTE: 
Wavelengths and power 
settings are factory preset and cannot 
be changed manually by the user. 
 
 
 
 
 
 
 
 
 
 
10 
The system performs an automatic system self-test each time the Examination screen is 
accessed for the first time after the system was switched on. 
11 
The transducer must remain in the magnetic probe holder fixed to the acquisition unit until 
the system self-test has been completed. If the ultrasound probe is not in the probe holder, 
the MSOT operator will make sure that it is placed there. 
12 
The staff will don safety goggles. The MSOT operator can 
control laser activation and deactivation via touch screen or 
keyboard in combination with the connected foot pedal. 
 
Hb, HbOmelanin 

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13 
The MSOT operator will hold the foot pedal until the self-test routine is finished. If the 
self-test is interrupted by releasing the foot pedal before the system self-test has been 
finalized, the test routine will need to be performed again. 
14 
A blue indicator in the signaling device on top of the 
system and in the front panel shows that the laser is ready 
for emission. 
 
 
 
15 
The MSOT operator will record the self-test completion on the case report form and note 
the Start scan time. 
NOTE:  THE FOOT PEDAL WILL BE RELEASED ONCE SELF-TEST ROUTINE IS COMPLETE. 
 
16 
The MSOT operator will tell the surgical team when the self-test routine has been 
successfully completed.  
17 
The MSOT operator will remove the probe from the magnetic 
holder to disinfect it using a germicidal disposable wipe (Sani-
Cloth AF3, PDI, Orangeburg, NY), ensuring that the entire surface 
of the probe is moistened with the disinfectant. 
 
18 
The MSOT operator will remove a second wipe from the package and repeat the 
disinfection process, ensuring that the entire surface of the probe is moistened with the 
disinfectant and allowed to dwell for at least 3 minutes.  
NOTE: MSOT Operator will note dwell time on Imaging Case Report Form (Appendix A). 
 
19 
After the 3-minute exposure time of the disinfectant, the MSOT operator will dry the probe 
surface with a commercially available, single-use, lint-free, low-germ towel. 
20 
A member of the surgical team will cover the handheld probe with a single-use ultrasonic 
probe drape (US.10.150.080, SonoGuard, Promecon GmbH, Hamburg, Germany), which 
will be the only part of the Acuity device to make contact with the patient, see (Figure 3). 
 
21 
Once time-out procedures have been completed, the surgical team will prepare the 
patient/region of interest on the skin for the initial MSOT scanning. 
22 
The skin surface will be cleaned following standard procedures. The skin will be dry 
without any alcohol pooling per manufacturer instructions, then ultrasound gel (#03-034, 
Aquasonic clear ultrasound gel, Parker Laboratories, Inc., Fairfield, NJ) will be applied to 
the area to be scanned. 
23 
The surgeon will place the imaging probe on the prepped skin area to be imaged BEFORE 
the laser is activated to avoid excess laser radiation. 
24 
Once the surgeon has the probe in place, the MSOT operator will press the foot pedal to 
activate the laser radiation for imaging. The MSOT operator will communicate with the 
surgeon as an image is acquired and coordinate the timing of the laser activation for 
additional scans. Release of the foot pedal will immediately shut down the laser system 
and stop the imaging procedure. 
25 
The MSOT operator will record Scan comments on the screen to be saved for each image 
acquired in the image sequence.  
NOTE:  Total number of images acquired in the image sequence will be recorded on the Imaging Case Report Form 
(Appendix A). 

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26 
A yellow indicator in the signaling device on top of the 
system and in the front panel shows that laser light is 
emitted through the aperture of the handheld probe. Keep 
the aperture of the probe on the region of interest or 
towards the floor. NEVER direct the aperture of the probe 
towards eyes while this indicator is on. 
Once the foot pedal is released laser emission is stopped 
and the laser emission indicator will turn off. 
 
 
27 
The patient will undergo surgery for resection of the identified volume of tissue according 
to standard care. 
28 
After tumor or lymph node removal and incision has been closed, the area of interest will 
be reimaged by MSOT to obtain post-surgery images in the operating room by repeating 
steps 16 through 23. 
29 
An estimate of the percent amount of tumor remaining will be estimated by algorithms 
within in the MSOT computer programming for investigational use only. 
30 
The resected tumor will be taken to Pathology for volume assessment and pathological 
identification per medically established diagnostic standard care. 
31 
Data from the MSOT imaging is for research purposes only and not for treatment, and 
digital images will be kept on the MSOT computer, which will be stored in a secured room 
accessible only to study personnel. 
32 
Key parameters will be recorded on (Appendix A and B). 
7.2 
Duration of Follow Up 
 
Subjects will be actively enrolled only for the duration of pre-op through 
surgical recovery, and until post-op discharge unless any study-related 
adverse event occurs. Medical record review will occur up to 6 months to allow 
for follow-up pathology assessments to be gathered and provide information 
to allow for analyses to be performed as previously described. 
7.3 
Criteria for Removal from Study 
 
Patients can request to withdraw from the study at any time. Should patients 
consent to the study, but have tumor progression that prevents them from 
being a surgical candidate within 30 days, these patients may be removed at 
the Investigator’s discretion. There is no reason to remove a patient 
otherwise from the study. 

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8.0 
Adverse Events List and Reporting Requirements 
An adverse event (AE) is any untoward medical occurrence in a patient or clinical 
investigation subject administered an investigational product, and which does not 
necessarily have to have a causal relationship with this treatment. An AE can 
therefore be any unfavorable and unintended sign (including an abnormal 
laboratory finding, for example), symptom, or disease temporally associated with 
the use of a medical product, whether or not considered related to the medical 
product. All AEs are assessed by the center for severity, causality, outcome, 
seriousness, and if unanticipated. 
8.1 
Adverse Event List for MSOT  
 
A slight, reversible reddening and temperature increase of sensitive skin. 
All adverse events and expected treatment effects, including mild events, 
will be monitored throughout the study until discharge.  
8.2 
Adverse Event Characteristics 
 
• 
CTCAE term (AE description) and grade:  The descriptions and 
grading scales found in the revised NCI Common Terminology Criteria 
for Adverse Events (CTCAE) version 5.0 will be utilized for AE reporting. 
All appropriate treatment areas should have access to a copy of the 
CTCAE version 5.0. A copy of the CTCAE version 5.0 can be 
downloaded from the CTEP web site (http://ctep.cancer.gov). 
 
• 
‘Expectedness’: AEs can be ‘Unexpected’ or ‘Expected’ (see Section 
8.1 above) for expedited reporting purposes only.   
 
• 
Attribution of the AE: 
 
- 
Definite – The AE is clearly related to the study treatment. 
- 
Probable – The AE is likely related to the study treatment. 
- 
Possible – The AE may be related to the study treatment. 
- 
Unlikely – The AE is doubtfully related to the study treatment. 
- 
Unrelated – The AE is clearly NOT related to the study treatment. 
 
8.3 
Serious Adverse Events (SAE) 
 
Serious is defined as an adverse event that results in the following outcome: 
• 
Death  
• 
Life-threatening event (i.e. an event that places the patient, in the 
view of the investigator, at immediate risk of death from the event as it 
occurred). This does not include an event that, had it occurred in a more 
severe form, might have caused death  

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• 
Requires or prolongs hospitalization  
• 
Results in persistent or significant disability/incapacity  
• 
Congenital anomaly/birth defect.  
• 
Important medical event that, based upon appropriate medical 
judgement, may jeopardize the patient, and may require medical or 
surgical intervention to prevent one of the outcomes listed above. 
 
8.4 
Unanticipated Adverse Device Effects Reporting Requirements 
 
Unanticipated Adverse Device Effect (UADE) means any serious adverse 
effect on health or safety or any life-threatening problem or death caused 
by, or associated with, a device, if that effect, problem, or death was not 
previously identified in nature, severity, or degree of incidence in the 
investigational plan or application (including a supplementary plan or 
application), or any other unanticipated serious problem associated with a 
device that relates to the rights, safety, or welfare of subjects.  
 
A medical review shall immediately occur with all reported Unanticipated 
Adverse Device Effects (UADE). The UADE shall be assessed for an 
unreasonable risks’ determination. If an unreasonable risk has been 
determined, then the trial or the parts that cause the risk shall be stopped 
between 5-15 working days after making the determination.  
 
8.5 
Reporting of Unanticipated Problems, Adverse Events or 
Deviations 
 
Any unanticipated problems, serious and unexpected adverse events, 
deviations or protocol changes will be promptly reported by the principal 
investigator to the IRB and sponsor or appropriate government agency, if 
appropriate and per institutional guidelines. 
 
A description of all AEs and UADEs will be recorded on REDCap. All 
UADEs should be reported using the Sponsor Form (UADE Reporting 
Form) and MedWatch3500A to the following email or fax within 24 hours of 
the site notification of the event:  
 
Email: SCCIITreporting@ouhsc.edu or  
Fax: 1-405-271-1416  
 
In compliance with FDA regulations 21 CFR 812.150, any UADEs occurring 
during the investigation should be reported to the sponsor and reviewing 
IRB within 10 days of first notice of the event. The sponsor will conduct an 

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evaluation of the UADE and report the results of such evaluation as an 
UADE Report to FDA, all reviewing IRBs, and participating investigators 
within 10 working days after the sponsor first receives notice of the effect.  
 
An UADE Report will consist of a completed MedWatch 3500A form and a 
cover letter that provides an analysis of the event. Thereafter the sponsor 
shall submit such additional reports concerning the effect as per requests 
from FDA. 
 
8.6 
Post-imaging Procedural Events for MSOT 
 
The post-imaging procedural events for MSOT are those events that are 
expected and standard to post-imaging as for ultrasound and include slight, 
reversible reddening and temperature increase of sensitive skin for a 
duration of ≤ 24 hours post-surgery. All unexpected events shall be 
captured. Post-imaging procedural events that occur in increased frequency 
and severity over what should normally occur in the medical opinion of the 
investigator will be considered adverse events (AE). AEs will be reported. 
9.0 
Data and Safety Monitoring 
Safety oversight will be performed by Stephenson Cancer Center’s (SCC) internal Data 
and Safety Monitoring Committee (DSMC). The DSMC is composed of individuals with 
the appropriate expertise in adult and pediatric hematology and medical oncology, 
radiation oncology, translational and correlative science, pharmacy, nursing and 
biostatistics. The DSMC operates under the rules of an approved data safety monitoring 
plan which complies with the National Cancer Institute (NCI) guidelines published as 
Essential Elements of a Data and Safety Monitoring Plan for Clinical Trials Funded by 
NCI as of January 2005 and the “NIH Policy for Data and Safety Monitoring,” NIH Guide 
for Grants and Contracts, http://grants.nih.gov/grants/guide/notice-files/not98-084.html. 
The Data Safety Monitoring Committee is charged with oversight of participant safety, 
study conduct and the validity and integrity of data for clinical trials at SCC. While the 
focus of the DSMC is to monitor interventional investigator-initiated trials (IITs) that are 
not subject to external monitoring, it has the authority to monitor any SCC protocol when 
potential concerns are identified. The DSMC also has the authority to suspend or close a 
study until the principal investigator addresses any issues that may cause harm or 
increase risks to subjects.  The DSMC reports all findings to the Institutional Review Board 
(IRB).  

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Under the direction of the DSMC chair, a full board meeting is convened on a quarterly 
basis to review the accumulated safety data, accrual information, and additional 
information as stated in the DSMC plan. 
9.1 
DSMC Auditing 
 
In addition to monitoring, the DSMC oversees an internal auditing process to ensure 
subject safety and data quality. All cancer-related clinical trials active at the SCC are 
eligible for audit; however, priority is placed on those clinical trials that are not monitored 
or audited by an outside entity. If an external entity conducts an audit of a clinical trial at 
the SCC, then the findings of that audit are reported to the DSMC, either through the 
formal audit report provided by the external auditing entity, if available, or from the PI, 
who will report any findings communicated during the audit process. 
 
9.2 
Data Handling and Record Keeping 
9.2.1 Data Quality Assurance 
 
Stephenson Cancer Center (SCC) will be responsible for clinical 
monitoring of all data for this study. 
 
9.2.2 Electronic Database and Case Report Forms 
 
The Principal Investigator and designated team will develop a study-specific 
electronic database in REDCap and study-specific case report forms for 
study data entry.  All study data will be recorded into the REDCap database 
sponsored by OU-SCC and stored in a 21 CFR 11-compliant database. 
Appendix A and B will be paper based and later scanned into REDCAP.  
Only the Investigator and assigned research staff will have access to the 
study data.  All case report forms will be available to Sponsor, IRB or 
regulatory authorities in event of an audit or inspection.   
 
9.2.3 Data Disclosure and Subject Confidentiality 
 
Subject medical information obtained as a result of this study is considered 
confidential. Disclosure to third parties other than those noted below is 
prohibited. All reports and communications relating to subjects in this study 
will identify each subject only by their initials and number. Medical 
information resulting from a subject’s participation in this study may be given 
to the subject’s personal physician or to the appropriate medical personnel 
responsible for the subject’s welfare. Data generated as a result of this 
study are to be available for inspection on request by FDA or other 
government regulatory agency inspectors, the SCC clinical trial office 
auditors and monitors, Office of Compliance, and the Institutional Review 
Board (IRB). 

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All evaluation forms, reports, and other records that leave the site will be 
identified by a coded number, and/or initials to maintain subject 
confidentiality. All study records will be kept in a locked file cabinet or other 
secured area. All computer entry and networking programs will be 
identifiable only by coded numbers and/or initials. Subject personal medical 
information may be reviewed by representatives of the Sponsor, IRB, or 
regulatory authorities in the course of monitoring the progress of the clinical 
trial. Every reasonable effort will be made to maintain such information as 
confidential. 
9.2.4 Record Retention 
 
In accordance with 21 CFR 812.140, the Sponsor and Investigator will retain 
all research documents relating to the participation in the clinical trial.  
These documents must be retained for a period of two years after the date 
on which the investigation is terminated or completed, or the date that the 
records are no longer required for purposes of supporting a premarket 
approval application, a notice of completion of a product development 
protocol, a humanitarian device exemption application, a premarket 
notification submission, or a request for De Novo classification. 
 
If the transfer of records custody during the retention period is accepted by 
any other person, notice of transfer must be given to FDA not later than 10 
working days after transfer occurs. 
 
9.3 
Study Monitoring 
 
All aspects of the study will be carefully monitored at periodic intervals throughout 
the study per FDA/ICH “Guidance for Industry – E6 Good Clinical Practice: 
Consolidated Guidance” dated April 1996. All Case Report Forms (CRFs) will be 
up to 100% source verified against corresponding source documentation (e.g., 
office and clinical laboratory records) for each subject. The monitoring visits 
provide the PI with the opportunity to evaluate the progress of the study, to verify 
appropriate 
consent form procedures, and to verify the accuracy and 
completeness of CRFs, to resolve any inconsistencies in the study records and to 
assure that all protocol requirements, applicable FDA regulations, other 
requirements, and Investigator’s obligations are being fulfilled. 
Furthermore, this study will fall under the purview of the Stephenson Cancer 
Center Data Safety Monitoring Committee (DSMC). 
10.0 Statistical Considerations 
10.1 Analysis of Primary Objective 
 
 

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10.1.1 The primary objectives are to estimate the rate of adverse events 
and measure the skin temperature change following MSOT imaging 
both pre- and post-surgery.  The adverse events will be estimated by 
grade, type and relatedness.  The first analysis will pool all grade 3 
unexpected events that are possibly, probably or definitely related to 
MSOT together and then construct a 95% exact Clopper-Pearson 
binomial 
confidence 
interval 
for this estimate. Additionally, 
measurement of skin temperature will be evaluated where skin 
temperature greater than 44oC will be recorded13.  A 95% exact 
Clopper-Pearson binomial confidence interval for the evaluation of 
skin temperature will be conducted 
10.2 Analysis of Secondary Objective 
 
10.2.1 There are three secondary outcomes of interest, two of these will be 
based upon a positivity (yes/no) (secondary outcomes 1 and 3) and 
one of these will assess on a continuous scale tumor size/ volume 
(secondary outcome 2).  In each of these analyses, we will compare 
the information gathered from the MSOT approach (size, positivity 
tumor (yes/no), positivity of dye (yes/no)) for each patient and then 
compare these measures to those that are available from information 
gathered in the standard of care approach (i.e., from pathology 
reports after resection). 
 
For continuous outcomes we will examine three characteristics 
based on comparing the MSOT and standard of care (“gold 
standard”) assessments.  These include: 1) the mean difference in 
the measure (i.e., tumor volume or size), 2) the correlation between 
the measures taken using MSOT or standard of care; and 3) the 
comparison of variability of the measures from the two approaches 
(MSOT and standard of care).  For the first approach we will calculate 
a paired t-test, for the second we will estimate a Pearson correlation 
and for the third we will perform an F-test to compare the equality of 
variances.  With the expanded sample size of 50 patients we can 
detect a difference in values of volume (or size) equal to 0.4 standard 
deviations or less of the measure of difference in tumor volumes (or 
size) with 80% power using a paired t-test with alpha=0.05 (2-sided).  
In addition, there is 80% power to detect a correlation of 0.38 (or 
larger) with an alpha=0.05 (2-sided).  Finally, there is 80% power to 
detect a ratio of 2.24 (or larger) when comparing variances between 
the two groups (note: this ratio corresponds to a ratio of 1.5 on the 
standard deviation scale).   
 
If the study is terminated early with 45 patients scanned, we can still 
detect a difference in values of volume (or size) equal to 0.43 
standard deviations or less of the measure of difference in tumor 
volumes (or size) with 80% power using a paired t-test with 

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alpha=0.05 (2-sided).  In addition, there is 80% power to detect a 
correlation of 0.40 (or larger) with an alpha=0.05 (2-sided). 
Additionally, there is 80% power to detect a ratio of 2.35 (or larger) 
when comparing variances between the two groups (note: this ratio 
corresponds to a ratio of 1.53 on the standard deviation scale) 
 
For binary outcomes (positivity and presence of dye), we will 
estimate the MSOT approach and the corresponding 95% Clopper-
Pearson exact binomial confidence interval.  Accuracy is defined as 
the number of patients where the MSOT and standard of care 
approach are in agreement (i.e., both are positive or both are 
negative).  In addition, we can examine other characteristics of the 
two approaches including sensitivity, specificity, positive predictive 
value and negative predictive values. 
10.3 Power and Sample Size  
 
10.3.1 We will initially accrue 10 patients for safety analysis regarding the 
use of the MSOT device pre- and post-surgery. If there are no grade 
III-V AEs observed in the patients as defined in CTCAE v5.0 per 
Section 8.2, we will submit an IDE supplement to request an 
enrollment expansion for an additional 40 patients, for a total of 50 
patients. Up to an additional 55 patients will be accrued (to account 
for patient dropouts) to enroll 40 patients. Thus, a total of 50 patients 
will be enrolled and scanned (10 patients in the initial analysis and 
40 patients following the interim analysis) in order to gather sufficient 
data to evaluate the proportion of patients with adverse events that 
may be associated with the MSOT approach.  The MSOT approach 
is not expected to be related to any adverse events except for a 
possible slight, reversible reddening and temperature increase of 
sensitive skin.  Nevertheless, this study will allow us to estimate the 
proportion of unexpected adverse events of grade 3 or higher that 
are possibly, probably, or definitely related to the MSOT approach.  
With 50 patients, the upper bound of a 95% Clopper-Pearson exact 
confidence interval will be less than 20% if the observed rate of 
adverse events is 8% or less (4 out of 50 or less).  We expect that 
the actual value will be 0 out of 50 – and if this is observed the upper 
bound of this confidence interval will be 7.1% which would provide a 
useful upper bound for the potential of adverse events.  
 
The study will be terminated early, after 45 patients in total are 
enrolled and scanned, if 3 or fewer AEs are observed in the patients 
scanned by the MSOT device, based on the assumption that the 
upper limit of the 95% Clopper-Pearson exact confidence interval will 
be less than 20%.  The upper limit of the 95% Clopper-Pearson exact 
confidence interval will be 7.9%, 11.7%, 15.1% or 18.3% respectively 
for 0, 1, 2, or 3 observed AEs. 

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10.3.2 Interim analysis: This study will initially accrue 10 patients to identify 
any potential grade III or IV AEs defined in CTCAE v5.0 per Section 
8.2. As yet, no study using MSOT in over 600 patients has observed 
any grade III or IV AEs2,3,14,15. An interim analysis will be conducted 
after the first 10 patients. If there are no grade III or IV AEs observed 
in the patients, we will submit an IDE supplement with preliminary 
safety data, to request an enrollment expansion to 50 patients. If any 
grade V AEs occur during the investigation, the FDA will be notified 
immediately and the study halted for further accrual until discussed with 
FDA. 
10.4 Estimated Accrual Rate  
 
10.4.1 The estimated accrual rate is 10-20 patients per month after the 
initial accrual of 10 patients. 
 
10.5 Estimated Study Length  
 
10.5.1 Subjects will be actively enrolled only for the duration of pre-op 
through surgical recovery, and until post-op discharge unless any 
study-related adverse event occurs.  Medical record review will occur 
up to 6 months to allow for follow-up pathology assessments to be 
gathered and provide information to allow for analyses to be 
performed as previously described. 
 
11.0 ETHICAL AND REGULATORY CONSIDERATIONS 
 
11.1 Ethical Conduct of the Study 
 
The study will be conducted in full conformance with the ICH E6 guideline for Good 
Clinical Practice and the principles of the Declaration of Helsinki in addition to the 
requirements of the ICH E2A guidelines. This study will also comply with U.S. FDA 
regulations under a U.S. Investigational Device Exemption (IDE) application in addition 
to local, state, and federal laws. 
 
11.2 Informed Consent 
 
The informed consent document will be in compliance with ICH GCP, local regulatory 
requirements, and legal requirements. The informed consent document used in this study, 
and any changes made during the course of the study, will be prospectively approved by 
the IRB of record. 

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The investigator must ensure that each study patient is fully informed about the nature 
and objectives of the study and possible risks associated with participation. The 
investigator, or a person designated by the investigator, will obtain informed consent from 
each patient before any study-specific activity is performed. The study site will retain the 
original of each patient’s signed consent document. A copy of the informed consent 
document must be provided to the subject or the subject's legally authorized 
representative.  If applicable, it will be provided in a certified translation of the local 
language. 
 
 
 

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References 
 
1. 
McNally LR, Mezera M, Morgan DE, et al. Current and Emerging Clinical Applications of 
Multispectral Optoacoustic Tomography (MSOT) in Oncology. Clin Cancer Res. 
2016;22(14):3432-3439. 
2. 
Diot G, Metz S, Noske A, et al. Multispectral Optoacoustic Tomography (MSOT) of 
Human Breast Cancer. Clinical cancer research : an official journal of the American 
Association for Cancer Research. 2017;23(22):6912-6922. 
3. 
Knieling F, Neufert C, Hartmann A, et al. Multispectral Optoacoustic Tomography for 
Assessment of Crohn's Disease Activity. The New England journal of medicine. 
2017;376(13):1292-1294. 
4. 
Waldner MJ, Knieling F, Egger C, et al. Multispectral Optoacoustic Tomography in 
Crohn's Disease: Noninvasive Imaging of Disease Activity. Gastroenterology. 
2016;151(2):238-240. 
5. 
Bhutiani N, Kimbrough CW, Burton NC, et al. Detection of microspheres in vivo using 
multispectral optoacoustic tomography. Biotechnic & histochemistry : official publication 
of the Biological Stain Commission. 2017;92(1):1-6. 
6. 
Zeiderman MR, Morgan DE, Christein JD, Grizzle WE, McMasters KM, McNally LR. 
Acidic pH-targeted chitosan capped mesoporous silica coated gold nanorods facilitate 
detection of pancreatic tumors via multispectral optoacoustic tomography. ACS Biomater 
Sci Eng. 2016;2(7):1108-1120. 
7. 
Becker A, Masthoff M, Claussen J, et al. Multispectral optoacoustic tomography of the 
human breast: characterisation of healthy tissue and malignant lesions using a hybrid 
ultrasound-optoacoustic approach. Eur Radiol. 2018;28(2):602-609. 
8. 
Stoffels I, Morscher S, Helfrich I, et al. Metastatic status of sentinel lymph nodes in 
melanoma determined noninvasively with multispectral optoacoustic imaging. Sci Transl 
Med. 2015;7(317):317ra199. 
9. 
Kimbrough CW, Khanal A, Zeiderman M, et al. Targeting Acidity in Pancreatic 
Adenocarcinoma: Multispectral Optoacoustic Tomography Detects pH-Low Insertion 
Peptide Probes In Vivo. Clin Cancer Res. 2015;21(20):4576-4585. 
10. 
Bhutiani N, Grizzle WE, Galandiuk S, et al. Noninvasive Imaging of Colitis Using 
Multispectral Optoacoustic Tomography. Journal of nuclear medicine : official 
publication, Society of Nuclear Medicine. 2017;58(6):1009-1012. 
11. 
Gurka MK, Pender D, Chuong P, et al. Identification of pancreatic tumors in vivo with 
ligand-targeted, pH responsive mesoporous silica nanoparticles by multispectral 
optoacoustic tomography. J Control Release. 2016;231:60-67. 
12. 
Hudson SV, Huang JS, Yin W, et al. Targeted noninvasive imaging of EGFR-expressing 
orthotopic pancreatic cancer using multispectral optoacoustic tomography. Cancer Res. 
2014;74(21):6271-6279. 
13. 
Yarmolenko PS, Moon EJ, Landon C, et al. Thresholds for thermal damage to normal 
tissues: an update. Int J Hyperthermia. 2011;27(4):320-343. 
14. 
Maximilian J. Waldner, Ferdinand Knieling, Cornelia Egger, et al. Multispectral 
optoacoustic tomography in Crohn’s disease: Non-invasive imaging of disease activity. 
Gastroenterology. 2016. 
15. 
Stoffels I, Morscher S, Helfrich I, et al. Metastatic status of sentinel lymph nodes in 
melanoma determined noninvasively with multispectral optoacoustic imaging. Science 
Translational Medicine. 2015;7(317):317ra199-317ra199. 
 

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Appendix A – Imaging and Tissue Data Collection Form 
 
 
Study Number: OU-SCC-MSOT 
 PID:  0 1 - __ __ __ 
 
Investigator: Lacey McNally, PhD 
 Surgery Date: __ __/__ __/__ __ 
 
1) Tumor or Lymph Node Assessments:    
 
a) Target tissue 
 
 Tumor    
 
 
 Lymph Node    
 
b) Standard of care method of imaging 
 
 MRI ______________tumor amount 
 
 PET ______________tumor amount 
 
 CT ______________tumor amount 
 
 Other: __________________________________ tumor amount 
 
c)  Did patient receive neoadjuvant therapy?  
 
 Yes 
   
 No   
 
d)  MSOT Image acquired?   
 
 Yes 
   
 No   
 
e) Was an indicator dye used during surgery? 
 
 Yes 
   
 No 
 
f) If an indicator dye used during surgery, please indicate which dye. 
 

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 Isosulfan Blue       
 IC Green  
2) Location of Imaging 
 
a) Please circle the location of the imaging (below) and provide further description as necessary 
__________________________________________________________________________________
__________________________________________________________________________________
__________________________________________________________________________________ 
 
 
b) Erythema post-MSOT Imaging (RTOG) 
 
i. 
Time after imaging: (hr: min) _______________ 
 
ii. 
Grade (RTOG) 
 0        No erythema present 
 1  
Follicular, faint or dull erythema, epilation, dry desquamation, decreased 
sweating 
 2    
Tender or bright erythema, patchy moist desquamation, moderate edema 
 3  
Confluent, moist desquamations other than skin folds, pitting, edema 
 4 
Ulceration, hemorrhage, necrosis 
 
3) Clinical Pathology Assessment 
 
a)   Date or diagnosis (month / year):  
 
 
b)   Positive tumor tissue: 
 
 
 
https://anatomyclass123.com / 

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 Yes           
 No   
c)  For 
 Pancreatic    
 melanoma    
 breast     
 HNSCC   
 ovarian (check one) indicate stage  
 
     at diagnosis (TNM classification): 
 
 
 Tx    
 T0       
 Tis       
 T1      
 T2        
 T3       
 T4 
 
 
 Nx    
 N0       
 N1      
 N2        
 N3        
 
 Mx    
 M0       
 M1      
 
 
d) Tumor subtype (if applicable) _______________________________________________________ 
 
4) Skin temperatures 
 
 
 
5) Patient Monitoring (***please note that patients must be monitored daily for adverse events 
until discharge). Please answer yes or no if Erythema at grade III or higher or change in skin 
pigmentation occurs.  
 
 Yes           
 No   
 
____________Post surgery 
 
 
___________ 8 day 
               ____________1 day                                                  ____________9 day 
____________2 day                                                  ____________10 day 
____________3 day                                                  ____________11 day 
____________4 day                                                  ____________12 day 
____________5 day                                                  ____________13 day 
____________6 day                                                  ____________14 day 
               ____________7 day                                                  ____________15 day 
              
             
               Discharge date: _______________ 
 
Pre-Surgery MSOT imaging 
Pre-imaging Skin 
Temperature (oC) 
 
Time  
 
Post-imaging Skin 
Temperature (oC) 
 
Time 
 
Post-Surgery MSOT imaging 
Pre-imaging Skin 
Temperature (oC) 
 
Time  
 
Post-imaging Skin 
Temperature (oC) 
 
Time 
 

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6) Adverse event. Please answer yes or no if Erythema at grade III or higher or change in skin 
pigmentation occurs. 
 
 
 Yes           
 No   
 
If yes, please fill out details below: 
 
____________ Erythema                ________ Time   __________Date 
 
____________ Changes in skin pigmentation   ____________   Date 
 
 
7)  MSOT Scan Times: 
 
MSOT Scan Time (hr: mm) 
Select the preset Hb, HbO, IsoBlue, 
and/or ICG for wavelengths settings  
 
 Check when complete 
 
Pre-Surgery 
Scan Start Time: 
 
Scan End Time:  
 
Total Scan time:  
 
Post-Surgery 
Scan Start Time: 
 
Scan End Time:  
 
Total Scan Time: 
 
TOTAL scan time 
(Pre-Surgery+ Post-Surgery Scan times)  
 
 
 
 
 
Treating Physician Signature: ______________________ Date: ___ ___ / ___ ___ / ___ ___ 
 
PI Signature: ____________________________                  Date: ___ ___ / ___ ___ / ___ ___ 
 
 

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Appendix B – Image Analysis Form 
 
 
Study Number: OU-SCC-MSOT 
 PID:  0 1 - __ __ __ 
 
Investigator: Lacey McNally, PhD 
 Surgery Date: __ __/__ __/__ __ 
 
In vivo MSOT Size/Volume Assessment:  
 
Tumor depth (mm): 
MSOT parameters 
Hb, a.u. 
 
Pre-Surgery:                           Post-Surgery: 
Hbtotal 
 
Pre-Surgery:                           Post-Surgery: 
HbO 
Pre-Surgery:                           Post-Surgery: 
Isosulfan Blue 
 Yes                                              Tissue:  
   
 No                                               Value: 
IC Green 
 Yes                                              Tissue:  
   
 No                                               Value: 
 
Duration of disease control: The duration of time from the date measurement criteria are first met for 
irCR, irPR, or irSD, until the first date that irPD is confirmed by independent radiology review or death, 
whichever comes first. 
 
 
Treating Physician Signature: ______________________ Date: ___ ___ / ___ ___ / ___ ___ 
 
PI Signature: ____________________________                  Date: ___ ___ / ___ ___ / ___ ___