This free webinar hosted by Scintica Instrumentation introduced participants to some of the basics of high frequency ultrasound imaging and reviewed the most common preclinical research applications for this exciting technology. Our presenter, Ms. Tonya Coulthard, discussed the basic principles of ultrasound imaging, presented sample images from various organs and tissues as a way of surveying common research applications, and introduced a few novel techniques that will be of interest to many researchers. Throughout the webinar, technical information, product specifications and sample images where shown from the Prospect T1 compact, high-frequency, preclinical ultrasound imaging system.

1. Tonya Coulthard, MSc.
Team Leader
Scintica Instrumentation
Phone: +1 (519) 914 5495
tcoulthard@scintica.com
Overview of High
Frequency Ultrasound
Imaging and a Survey of
Preclinical Applications

2. • Basics of Ultrasound Imaging
• Prospect T1 System Overview
• Key Research Applications
Topics of Discussion

3. Basics of Ultrasound Imaging
• Understanding how ultrasound works
• Review of acquisition modes

4. • Ultrasound is a non-invasive imaging technique which does not required the use of ionizing radiation, instead it
uses sound waves
• The transducer both sends and receives the ultrasound waves and the computer interprets the returned signal
into an image
Ultrasound Imaging
Note – these images were not taken
with the Prospect T1 system

5. • High-frequency ultrasound waves are necessary to resolve the small anatomical targets in preclinical research
• Compromise is shorter penetration depth
Ultrasound Imaging

6. • B-Mode images are a 2D grey scale representation of anatomical structures – still or cine loop images
• The sound waves cannot penetrate through air or bone
• Ultrasound gel, or some other liquid, is used to couple the transducer to the surface
Acquisition Modes – Brightness (B) Mode

7. • M-Mode displays moving structures along a single line
• As data is only taken along a single line, the sampling frequency is very high (1000 frames per second)
• The user defines the sample volume on a B-mode image
Acquisition Modes – Motion (M) Mode

8. • Doppler modes detect moving structures, i.e. blood
• Velocity and direction of the blood can be determined by the “Doppler Shift”
• Color/Power Doppler are displayed as an overlay on a B-Mode image, while Pulsed Wave Doppler is displayed as
a spectrogram
Acquisition Modes – Doppler Modes

9. • Microbubble contrast agents may be used to image the vasculature or targeted cellular markers; using non-
targeted or targeted microbubbles respectively
• Reference subtracted or harmonic imaging may be used to visualize the contrast agents
Acquisition Modes – Contrast Mode

10. Prospect T1 System Overview
• System components and standard configuration
• Add-on hardware and software components

11. Prospect T1 System Components

12. Prospect T1
System Components
• The Prospect T1 is the first tablet based high-frequency
ultrasound system specifically designed for preclinical imaging
of small animals
• System components:
• Tablet
• Probe
• Scanning Platform

13. 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
• 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
• The intuitive workflow and touch screen allow researchers to start
acquire images and generating data quickly

14. Prospect T1
System Components:
Probes
• Two 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

15. Prospect T1
System Components:
Scanning Platforms
• 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

16. 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

17. Prospect T1 System Add-Ons

18. Add-Ons:
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

19. Add-Ons:
Image Guided Needle Injection
Mount
• 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
E15.5 mouse
embryo
Adult mouse
myocardium

20. Add-Ons:
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

21. Add-Ons:
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

22. Key Research Applications
• Cardiovascular Research
• Cancer Biology
• Abdominal & Anatomical Imaging
• Developmental Biology
• Ophthalmology
• Other Animal Models – Zebrafish, Chick Embryos

23. Cardiovascular Research

24. Cardiovascular Research:
Mouse Systolic Function – B-Mode
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
IVS : interventricular septum
AO : aortic orifice
LA : left atrium

25. Cardiovascular Research:
Mouse Systolic Function – B-Mode;
Area Length Measurement (ALM)
• End diastolic volume; End systolic volume
• Stroke volume
• Ejection fraction
• Fractional area change (from short axis)
• Fractional shortening
• Left ventricular mass
• Left ventricular mass index

26. Cardiovascular Research:
Mouse Systolic Function – M-Mode

27. 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

28. Cardiovascular Research:
Mouse Systolic Function – PW
Doppler & B-mode
• 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

29. Cardiovascular Research:
Mouse Diastolic Function – Color
& PW Doppler
LV
R
V
L
A
R
A
MV
T
V
LV : left ventricle
RV: right ventricle
LA: left atrium
RA: right atrium
MV: mitral valve
TV: tricuspid valve Mitral Valve
Tricuspid Valve

30. Cardiovascular Research:
Mouse Diastolic Function – Color
& 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)

31. Cardiovascular Research:
Mouse Diastolic Function – Tissue
Doppler
• Peak velocity of E & A waves
• Isovolumic relaxation/contraction time (IVRT & IVCT)
• Ejection time
• Filling time

32. Cardiovascular Research:
Mouse Aortic Arch
RPA
IA
LCCA
LSCA
AAr
AAr : Aortic Arch
RPA : Right Pulmonary Artery
IA: Innominate Artery
LCCA : Left Common Carotid Artery
LSCA : Left Subclavian Artery
AAr
AAo
IA
LCCA
LSCA

33. Cardiovascular Research:
Rat Carotid Artery
Intima-Media Thickness

34. Cardiovascular Research:
Mouse Carotid Artery
ICA
ECA RCCA
Right Common Carotid Artery (RCCA)
Internal Carotid Artery (ICA)
External Carotid Artery (ECA)

35. Cardiovascular Research:
Mouse Peripheral Vasculature –
Color & PW Doppler
• Automatic peak analysis is used to help with
peripheral vascular measurements
• Systolic:Diastolic ratio
• Peak systolic velocity
• End diastolic velocity
• Resistive index
• Pulsatility index

36. 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 injected into the myocardium
to study the effect on myocardial infarction lesion size, for
example

37. Cancer Biology

38. Cancer Biology:
Early detection of tumors
Primary liver tumors (mouse)

39. Cancer Biology:
Surrounding Structures
Mouse Axillary Lymph Node
• Visualizing the surrounding structures helps
visualize additional changes which may occur
• This lymph node is normal, however they may be
enlarged and have an altered appearance with
advanced disease

40. Cancer Biology:
Tumor measurement
• Linear and area measurement tools allow tumor
sizing on 2D B-mode images
Orthotopic Liver Tumor - Mouse
Orthotopic Breast Tumor - Mouse

41. Cancer Biology:
Tumor measurement
• The 3D motor add-on can be used to acquire 3D B-
mode images
• Orthotopic breast tumor in a mouse
• Volume = 211mm3

42. Cancer Biology:
Tumor Perfusion
• Microbubble contrast agents are inject
intravenously to study perfusion
• Two types of contrast imaging are possible
• Reference subtracted – with green color overlay
applied on cineloop image (20 or 40MHz
probe)
• Harmonic imaging – microbubble specific
harmonic signal is detected (20MHz probe)

43. Abdominal & Anatomical

44. Abdominal & Anatomical:
Mouse Liver and Gallbladder
Liver
Liver vessels
Gallbladder

45. Abdominal & Anatomical:
Mouse Spleen and Pancreas
Spleen
Splenic vein
Pancreas
Spleen
Kidney

46. Abdominal & Anatomical:
Mouse Kidney
Kidney

47. Abdominal & Anatomical:
Mouse Abdominal Aorta

48. Abdominal & Anatomical:
Mouse Knee and Lower Leg
Tibiofemoral tendon
Synovial membrane
Cartilage
Patella
Muscle
Lower Leg

49. Abdominal & Anatomical:
Mouse Ovary
Ovary Ovary
Cyst

50. Developmental Biology

51. Developmental Biology:
Mouse Embryos
Embryonic heart and
neural tube _E9.5
Embryos_E7.5
Embryonic brain_E12.5
Embryonic spinal cord_E12.5
Embryonic head and
forelimb_E14.5

52. Developmental Biology:
Mouse Embryos – Color and PW
Doppler
Mitral Valve
Umbilical cord
Dorsal Aorta

53. Developmental Biology:
Mouse Embryos – M-Mode
Heart Ventricles

54. Developmental Biology:
Image Guided Needle Injection;
Mouse Embryo
• 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

55. Opthamology

56. Ophthalmology:
Anterior Structures and Retinal
Vasculature; mouse eyeEyelid
CorneaLens
Cornea
Ciliary
body

57. Ophthalmology:
3D Imaging; rat eye
Volume = 57.6mm3

58. Ophthalmology:
Anterior Structures; rabbit eye
Normal
Thickened Cornea

59. Other Animal Models

60. Other Animal Models:
Zebrafish
Gills
Fin
Eye
Spinal Cord
Ventricular Inflow
E
A

61. Other Animal Models:
Chick Embryo
5 Day
7 Day
7 Day

62. • Basics of Ultrasound Imaging
• Prospect T1 System Overview
• Key Research Applications
Topics of Discussion

63. Tonya Coulthard, MSc.
Team Leader
Scintica Instrumentation
Phone: +1 (519) 914 5495
tcoulthard@scintica.com
Q&A
SESSION:
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