Hands-on demonstration of the Prospect T1, high-frequency ultrasound system During this live demonstration we demonstrated the animal preparation steps and positioning for both mice and rats on the Prospect T1. We also showed the following cardiac views on both species: Long Axis, in B-Mode Short Axis, in B-Mode and M-Mode Pulmonary Artery, in B-Mode, Color Doppler, Pulsed Wave Doppler Apical 4-Chamber to Measure Mitral Valve Flow, in B-Mode, Color Doppler, Pulsed Wave Doppler, and Tissue Doppler We also examined the aortic arch, carotid arteries, abdominal aorta, and other abdominal organs throughout the demonstration.

1. Prospect T1
Live Demonstration
Compact, High-Frequency, Tablet-based
Small Animal Ultrasound System
Presenter:
Tonya Coulthard - tcoulthard@scintica.com
Manager, Imaging Division
Scintica

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Topics of Discussion
• Prospect T1 System Overview
• Pre-Recorded System Demonstration
• Example Images and Sample Measurements
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Prospect T1 System Overview
• System components and standard configuration
• Add-on hardware and software components
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Prospect T1 System Components
• The Prospect T1 is the first tablet based high-frequency ultrasound system
specifically designed for pre-clinical imaging of small animals
• System components:
• Tablet
• Probe
• Scanning Platform
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Prospect T1 System Components: Tablet
• The powerful tablet reduces the footprint of the system, taking up less lab
space, making it easy to move when necessary
• The intuitive workflow and touch screen allow researchers to start acquire
images and generating data quickly
• Multiple data formats exist for either still, cine loop, or RAW data storage
• Offline software analysis is possible to preserve time on the system for
imaging
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Prospect T1 System Components: Probes
• Three single element probes are available:
• 20 MHz (user selectable between 15-30 MHz)
• Primarily used for rat imaging, as well as harmonic contrast
imaging
• 40 MHz (user selectable between 30-50 MHz)
• Primarily used for mouse imaging, and superficial anatomical
targets in larger species like rats
• 50 MHz (user selectable between 30-50 MHz)
• Primarily used for superficial anatomical targets in both mice
and rats
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Prospect T1 System Components: Scanning Platform
• The platform is compact in design, again to limit the footprint of the system
• The scanning platform has been designed for ergonomical positioning of the
probe
• Animal beds have integrated heating, and ECG and respiratory monitoring
• Interchangeable beds are available for mice or rats
• Animal beds can be precisely adjusted in the X, Y, and Z axis
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Standard System Configuration
• Standard system configuration for mouse
• B-Mode
• M-Mode
• Pulsed Wave / Color / Power / Tissue Doppler
Mode
• Contrast Mode
• Comprehensive Measurement and Analysis
Tools
• Scanning Platform – with mouse bed
• 40 MHz probe
• Standard system configuration for rat
• B-Mode
• M-Mode
• Pulsed Wave / Color / Power / Tissue Doppler
Mode
• Contrast Mode
• Comprehensive Measurement and Analysis
Tools
• Scanning Platform – with rat bed
• 20 MHz probe
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Add-On Hardware: 3D Motor
• The 3D motor expands the capabilities of the Prospect T1 to acquire 3D B-
mode images
• Add-on includes the software analysis package to view the 3D images and
perform volume calculations
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Add-On Hardware: Image Guided Needle Injection
• The image guided needle injection mount integrates with probe
• Injections may be performed with a regular syringe and steel needle, or
pulled glass capillary needle
• Injections may be made into developing embryos, adult myocardium, or
abdominal/muscle targets
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E15.5 mouse
embryo
Adult mouse
myocardium

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Add-On Hardware: Shear Wave Elastography
• Shear wave elastography is used to quantify mechanical and elastic
properties of tissues
• The acoustic radiation force is generated by a push probe mounted on the
side of the imaging probe
• The software analysis generates a colored elastogram which is overlaid on
a B-mode image
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Add-On Hardware: Integrated Sonoporation
• Sonoporation is the controlled cavitation or bursting of microbubbles with
the intention of increasing the permeability of the cell membrane or to
open to blood brain barrier
• Sonoporation is performed by a secondary, non-imaging, probe directed
at the anatomical target
• Software integration and control of the sonoporation probe is included with
this add-on
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Key Research Applications
• Cardiovascular Research
• Cancer Biology
• Abdominal & Anatomical Imaging
• Developmental Biology
• Ophthalmology
• Other Animal Models – Zebrafish, Chick Embryos
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• Cardiovascular Research
• Systolic function
• Diastolic function
• Left Ventricle, Right Ventricle
• Aorta, Pulmonary Artery
• Mitral Valve, Tricuspid Valve
• Peripheral Vessels
• Image Guided Needle Injections

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Cardiovascular Research: Mouse Systolic Function
B-Mode
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Long Axis View
IVS
LV
LVPW
AO
LA
PM Mitral valve
Short Axis View
IVS LV
LVPW
LVAW
PM
LV : left ventricle
LVAW: left ventricular anterior wall
LVPW : left ventricular posterior wall
PM: papillary muscle
VS : interventricular septum
AO : aortic orifice
LA : left atrium

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Cardiovascular Research: Mouse Systolic Function
ECG Gated Kilohertz Visualization (EKV) Mode
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B-Mode Frame Rate = 30fps ECG Gated Kilohertz Visualization = 30fps

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Cardiovascular Research: Mouse Systolic Function
B-Mode; Area Length Measurement
• End diastolic; End systolic
volume
• Stroke volume
• Ejection fraction
• Fractional area change (from
short axis)
• Fractional shortening
• Left ventricular mass
• Left ventricular mass index
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Cardiovascular Research: Mouse Systolic Function
B-Mode; LV Volume Integral Technique
• End diastolic; End systolic
volume
• Stroke volume
• Ejection fraction
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Cardiovascular Research: Mouse Systolic Function
B-Mode; Teichholz Formula
• Can be done on either the long
or short axis B-mode image
• LV mass
• LV mass index
• Fractional shortening
• End diastolic volume; end
systolic volume
• Stroke volume
• Ejection fraction
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Cardiovascular Research: Mouse Systolic Function
B-Mode; Modified Simpson’s Rule
• Fractional shortening
• End diastolic volume; end
systolic volume
• Stroke volume
• Ejection fraction
• Fractional area change
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Cardiovascular Research: Mouse Systolic Function
M-Mode
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Cardiovascular Research: Mouse Systolic Function
M-Mode
• Can be done on either the long or
short axis M-mode image
• LV mass
• LV mass index
• Fractional shortening
• End diastolic volume; end
systolic volume
• Stroke volume
• Ejection fraction
• Cardiac output
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Cardiovascular Research: Mouse Systolic Function
B-Mode and PW Doppler – Ascending Aorta
• Stroke Volume is calculated as a
function of the Velocity Time Interval
(VTI) and vessel diameter
• Cardiac Output is simply stroke
volume x heart rate
• VTI can be manually or automatically
traced on the PW Doppler:
• Peak velocity
• Peak pressure gradient
• Mean velocity
• Mean pressure gradient
• Acceleration & Deceleration
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Cardiovascular Research: Mouse Systolic Function
B-Mode and PW Doppler – Pulmonary Artery
• Stroke Volume is calculated as a
function of the Velocity Time Interval
(VTI) and vessel diameter
• Cardiac Output is simply stroke
volume x heart rate
• VTI can be manually or automatically
traced on the PW Doppler:
• Peak velocity
• Peak pressure gradient
• Mean velocity
• Mean pressure gradient
• Acceleration & Deceleration
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Cardiovascular Research: Mouse Diastolic Function
Color and PW Doppler
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Mitral Valve
Tricuspid Valve
LV
RV
LA
RA
MV
TV
LV : left ventricle
RV: right ventricle
LA: left atrium
RA: right atrium
MV: mitral valve
TV: tricuspid valve

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Cardiovascular Research: Mouse Diastolic Function
Color and PW Doppler
• Acceleration rate of E wave
• Peak velocity of E & A waves
• Deceleration time of E wave
• E:A ratio
• Isovolumic relaxation/contraction
time (IVRT & IVCT)
• Ejection time
• Myocardial performance index
(Tei index)
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Cardiovascular Research: Mouse Diastolic Function
Tissue Doppler
• Peak velocity of E & A waves
• Isovolumic relaxation/contraction
time (IVRT & IVCT)
• Ejection time
• Filling time
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Cardiovascular Research: Mouse Aortic Arch
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RPA
IA
LCCA
LSCA
AAr
AAr
AAo
IA
LCCA
LSCA
AAr : Aortic Arch
RPA : Right Pulmonary Artery
IA: Innominate Artery
LCCA : Left Common Carotid Artery
LSCA : Left Subclavian Artery

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Cardiovascular Research: Mouse Carotid Arteries
B-Mode
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Cardiovascular Research: Mouse Carotid Arteries
M-Mode, Color and PW Doppler
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Vessel diameter through systole and diastole may be
measured using M-mode.
Color Doppler is used to visualize direction of
flow within vessels. Aliasing shows highest
velocity flow at bifurcation of carotid artery.
Resistive and pulsatility indices may be measured on
peripheral vessels using PW Doppler.

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Cardiovascular Research: Rat Systolic Function
B-Mode
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380g rat 600g rat

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Cardiovascular Research: Rat Carotid Arteries
B-Mode
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Cardiovascular Research: Image Guided Needle Injection – Adult
Mouse Myocardium
• Image guided injection may
be done into the myocardium
or other anatomical target
• Stem cells or other therapy
may be injected into the
myocardium to study the
effect on myocardial infarction
lesion size, for example
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• Cancer Biology
• Tumor detection
• 3D volume measurements
• Surrounding tissue investigation – i.e.
lymph nodes
• Blood flow monitoring
• Power Doppler
• Linear contrast agent imaging
• Non-linear contrast agent imaging

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Cancer Research – Preclinical Solid Tumor Models
• A number of different types of solid
tumor models are used commonly in
preclinical research:
• Cell line-derived models
• Patient Derived Xenograft (PDX)
models
• Environmentally induced models
• Genetically Engineered Mouse
(GEM) models
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Figure from Gengenbacher et al. Nature Reviews Cancer (2017) 17:751-765.

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Cancer Research – Tumor Detection and Monitoring
• Tumors of all types are visible,
whether subcutaneous or orthotopic
• Standard 2D B-mode imaging is used
to provide a greyscale image, tumors
often show up as a different
echogenicity than the surrounding
tissue
• 2D measurements can be done to
measure linear or area measurements
of tumor size
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Subcutaneous tumor model
Transgenic liver tumor model

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Cancer Research – Tumor Detection and Monitoring
• Complex tumor models can also be
investigated using ultrasound
• Normal tissues must be identified,
followed by the identification of
abnormal tissues
• Changes in nearby tissues may also
be investigated
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IP injection of ovarian tumor cells (SKOV-3)
Tumour
Stomach
Kidney Splenic Vein
Tumour
Intestine

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Cancer Research – 3D Tumor Volume Measurements
• 3D volume measurements can be made on any
visualized tumor using the 3D motor add-on
• Volume measurements may be used to follow
the same tumor over a longitudinal study to
monitor tumor progression or therapeutic
response
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Orthotopic Mammary Fat Pad Tumor (MDA-MB-231)
Volume = 263mm3

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Cancer Research – 3D Tumor Volume Measurements
• Complex tumor structures can be
visualized in 3D
• Longitudinal imaging to monitor
tumor progression or therapeutic
response
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IP injection of ovarian tumor cells (SKOV-3)
Tumor

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Cancer Research – 3D Tumor Volume Measurements
• Due to the complex nature of this
tumor model the images were
analyzed in VivoQuant
• Red Tumor = 4.7mm3
• Green Tumor = 16.0mm3
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IP injection of ovarian tumor cells (SKOV-3)

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Cancer Research – Surrounding Tissue Investigation
• Surrounding tissues may be
investigated, as can other tissues and
organs which may be affected
• Lymph nodes may show signs of
involvement, and may indicated
spread of the disease
• The spleen may be involved and
show an altered appearance or
larger size
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Lymph Node
Skeletal Muscle
Spleen

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Cancer Research – Blood Flow Monitoring
Power Doppler
• Surrounding tissues may be
investigated, as can other tissues and
organs which may be affected
• Lymph nodes may show signs of
involvement, and may indicated
spread of the disease
• The spleen may be involved and
show an altered appearance or
larger size
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Cancer Research – Blood Flow Monitoring
Linear Contrast Agent Imaging
• Microbubble contrast agents are
injected i.v. to study microvascular
perfusion
• Microbubbles are typically 2-3µm in
diameter, and mimic red blood cells
when they are non-targeted; may also
be targeted to bind to biomarkers
• Linear contrast agent imaging uses
reference subtraction to create a
green overlay, and is available on all
probes
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Cancer Research – Blood Flow Monitoring
Non-Linear (Harmonic) Contrast Agent Imaging
• Harmonic imaging is used to acquire
a more specific signal coming from
the microbubbles
• Using only the 20MHz probe, the
system will transmit at 20MHz, but
listen for the 1st harmonic – 40MHz. In
doing this, tissue signal is removed,
and only microbubbles are visualized
• Time vs. Intensity curves can be
created for numerous ROIs in both
modes
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• Abdominal & Anatomical
Imaging
• Organ visualization
• 3D volume measurements
• Surrounding tissue investigation, including
visualization of oedema
• Blood flow monitoring
• PW Doppler
• Color & Power Doppler
• Linear contrast agent imaging
• Non-linear contrast agent imaging

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Abdominal Imaging – Liver and Gallbladder (Mouse)
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Liver
Liver vessels
Gallbladder

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Abdominal Imaging – Spleen and Pancreas (Mouse)
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Spleen
Splenic vein
Pancreas
Spleen
Kidney

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Abdominal Imaging – Kidney (Mouse)
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Kidney

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Abdominal Imaging – Uterine Horn and Testis (Mouse)
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Bladder
Right Horn Left Horn
Testis
Caput Epididymis

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• Developmental Biology
• Confirmation of pregnancy
• Embryo counting
• Monitoring embryonic developmental
stages
• Monitoring cardiac function and flow
within the embryos
• Placental and umbilical cord flow
measurements
• Image guided needles injections into
externalized embryos

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Developmental Biology – Development (Mouse)
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Embryos_E7.5
Embryonic brain_E12.5
Embryonic spinal cord_E12.5
Embryonic head and
forelimb_E14.5
Embryonic heart and neural
tube _E9.5

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Developmental Biology – Cardiac Function in Pups (Mouse)
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Mitral Valve
Umbilical cord
Dorsal Aorta

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Developmental Biology – Image Guided Injection (Mouse)
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• Image guided injection may be done into a variety of anatomical targets
within the embryo
• The uterine horn is exposed from the dame and injections done into the
exposed embryos

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• Other Animal Models
• Same types of analysis can be completed
with all of the standard modes, as long as
the ultrasound signal can penetrate the
imaging subject
• Examples of animal models
• Zebrafish
• Chick embryo
• Other small animals
• Ultrasound gel or water may be used to
allow for imaging – i.e. zebrafish in their
tank water with an anesthetic

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Other Animal Models - Zebrafish
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Gills
Fin
Eye
Spinal Cord
Ventricular Inflow
E
A

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Other Animal Models – Chick Embyro
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5 Day
7 Day
7 Day

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