This will be a full day of live imaging where we will be examining the complimentary nature of three preclinical imaging modalities – bioluminescence, high-frequency ultrasound, and MRI. We will image the same tumor models on each of the imaging modalities; but will also have a normal mouse available to look at cardiovascular function using ultrasound, and brain anatomy using MRI.

1. Understanding the
Complementary Nature of
Imaging Modalities:
Live demonstration
October 1st, 2020
Tonya Coulthard – tcoulthard@scintica.com
Katie Parkins – kparkins@scintica.com

2. Images Acquired During The Live Demo
• Newton 7.0- Optical Imaging
• Prospect T1- Ultrasound Imaging
• M-series- MRI Imaging

3. COURTESY : Matthieu Germain , Nanobiotix/Curadigm
Newton 7.0 FT
In Vivo Optical Imaging System

4. Optical Imaging (Bioluminescence)
Signal Only Image Brightfield Image Overlay Image

5. IP metastasis model
(human ovarian
cancer)
Dual MFP Model
(human breast cancer)
Optical Imaging (Bioluminescence)

6. FLI Phantom @ 480nm Multiplex Phantom
Fluorescent
beads
Fluorescent
beads
Luminescent
beads
Optical Imaging (Bioluminescence)

7. • Lower binning improves resolution but lowers sensitivity; high binning leads to more signal but
more pixelated image quality; tail was covered to block signal from tracer (fluorescence)
Previously Acquired Images: Bioluminescence (Subcutaneous
Tumor)
Max greyscale: 8771
With hair; 4 minutes, 2x2 bin
Max greyscale: 26647
Shaved; 4 minutes, 2x2 bin
Binning= 2 Binning= 16

8. • Binning adjusts the sensitivity of the image, so much so that the acquisition time had to be
decreased from 4 minutes to 2 minutes to prevent saturation
Binning = 16Binning = 4
Previously Acquired Images: Bioluminescence (Orthotopic Lung
Tumors)

9. • These mice had an mCherry expressing tumor on the left side, and a GFP
expressing tumor on the right side; on both mice the GFP tumors were not visible
Previously Acquired Images
– mCherry Tumor on Left , GFP on Right

10. Prospect T1
Compact, High-Frequency, Tablet-based System
Small Animal Ultrasound

11. Tumor Imaging – 2D B-Mode Imaging
• Tumor identification, and basic linear and area
measurements
• Investigation of surrounding structures
• Longitudinal imaging to monitor tumor
progression or therapeutic response
Orthotopic Mammary Fat Pad
Tumor MDA-MB-231
• Power Doppler (not shown during the demo due to a technical issue with the demo system)
can be used to assess tumor vasculature using endogenous signal within the tissue
• Contrast Imaging (linear and subharmonic - not shown during the demo as no contrast
agent was available) can be used to assess tumor microvasculature using microbubbles

12. Tumor Imaging – 2D B-Mode Imaging
• The complex nature of tumors can be investigated using B-Mode imaging, as well as surrounding
structures
IP Injection of Ovarian Tumor Cells
SKOV-3
Tumor
Stomach
IP Injection of Ovarian Tumor Cells
SKOV-3
Kidney Splenic Vein
TumorIntestine

13. Tumor Imaging – 3D B-Mode Imaging
• 3D volume measurements
• Longitudinal imaging to monitor
tumor progression or therapeutic
response
Volume = 263mm3
Orthotopic Mammary Fat Pad
Tumor MDA-MB-231

14. Tumor Imaging – 3D B-Mode Imaging
• 3D volume measurements
• Complex tumor structures
can be visualized
• Longitudinal imaging to
monitor tumor progression or
therapeutic response
IP Injection of Ovarian Tumor Cells
SKOV-3
Tumor

15. Tumor Imaging – 3D B-Mode Imaging
• Due to the complex nature of this tumor model
the images were analyzed in VivoQuant
• Red Tumor = 4.7mm3
• Green Tumor = 16.0mm3

16. Cardiovascular Imaging – Par aster nal Long Axis View of LV
B-Mode Frame Rate = 30fps ECG Gated Kilohertz Visualization = 30fps

17. Cardiovascular Imaging – Par aster nal Long Axis View of LV

18. Cardiovascular Imaging – Pulsed Wave Doppler
Pulmonary Artery Mitral Valve

19. Cardiovascular Imaging – Left Carotid Artery
• 3D imaging is possible on blood vessels
• Here VivoQuant was used to outline both the left and right
carotid arteries

20. Cardiovascular Imaging – Color Doppler
• Color Doppler (not shown during the demo due to a technical issue with the demo system) can be used to identify and assess
blood flow velocity and direction within the heart and surrounding vasculature

21. Cardiovascular Imaging – Measurements
• Systolic cardiac function can be measured from an M-mode image
• Multiple instances of each measurement can be made, with the software outputting the individual values, average and standard deviation into a .csv file format
Measurement Unit
Value
Average STD
1 2 3
Heart Rate BPM 351 351 351 351 0
LVAW;d mm 0.99 1.07 0.99 1.01 0.04
LVID;d mm 3.36 3.36 3.49 3.41 0.06
LVPW;d mm 1.33 1.18 1.16 1.22 0.08
LVAW;s mm 1.8 1.58 1.63 1.67 0.09
LVID;s mm 1.78 1.88 1.88 1.85 0.05
LVPW;s mm 1.67 1.69 1.58 1.65 0.05
Calculation Unit
Value
Average STD
1 2 3
FS % 47.13 43.95 46.01 45.7 1.32
EDV uL 46.19 46.19 50.56 47.64 2.06
ESV uL 9.41 10.94 10.94 10.43 0.72
EF % 79.62 76.32 78.37 78.1 1.36
LV mass mg 121.65 116.49 114.4 117.51 3.05
SV uL 36.77 35.25 39.62 37.21 1.81
CO mL/min 12.92 12.37 13.91 13.07 0.63
• A full compliment of analysis options are available on the Prospect T1, a few are shown here

22. Cardiovascular Imaging – Measurements
• Systolic cardiac function can be measured from 2D B-Mode images by measuring the change in length in the long axis, and change in area in the short axis
• If multiple instances of each measurement were made then the .csv file would contain each value along with the average and standard deviation
Measurement Unit Value 1
Epi. Area;d mm2 25.82
Endo. Area;d mm2 9.64
LV Length;d mm 8.02
Epi. Area;s mm2 21.35
Endo. Area;s mm2 4.02
LV Length;s mm 7.04
Weight(ALM) gm 25
Calculation Unit Value 1
LVM(ALM) mg 138.67
LVMI(ALM) g/m2 18.02
LV EDV(ALM) uL 64.43
LV ESV(ALM) uL 23.58
SV(ALM) uL 40.84
EF(ALM) % 63.39
FS(ALM) % 35.44
Endo FAC(ALM) % 58.32

23. Cardiovascular Imaging – Measurements
• Diastolic cardiac function can be measured from the mitral valve inflow using PW Doppler and measuring the various velocity and time points
• If multiple instances of each measurement are made then the .csv file contains each value along with the average and standard deviation
Measurement Unit
Value
Average STD
1 2
A mm/s 254.89 181.1 218 36.89
Acc m/s2 38.43 28.72 33.57 4.85
DT ms 32.75 27.94 30.34 2.41
E mm/s 500.17 390.49 445.33 54.84
E/A 1.96 2.16 2.06 0.1
ET ms 45.28 39.01 42.14 3.13
IVCT ms 8.67 10.11 9.39 0.72
IVRT ms 18.78 17.82 18.3 0.48
MPI 0.61 0.72 0.66 0.05

24. M-Series
Compact, Self-Shielded, High Performance Permanent
Magnet
Small Animal MRI

25. Tumor Imag ing – Or thotop ic Mammar y Fat Pad Tumor
• MDA-MB-231 cell line
• T1 and T2 weighted image contrast helps
to identify anatomy; along with
pathological changes, including tumors
• 250µm in-plane resolution with 1mm slice
thickness
• Acquisition was around 4.5 minutes
T1 Weighted T2 Weighted

26. Tumor Imag ing – Or thotop ic Mammar y Fat Pad Tumor
• Tumor volume was found to be 273mm3
T1 Weighted T2 Weighted
MDA-MB-231

27. Tumor Imaging – IP Injected Ovarian Tumor Cells
• SKOV3 cell line, injected IP
• These tumors may form anywhere within the
peritoneal cavity
• Numerous tumors were located throughout
the abdomen
T2 WeightedT1 WeightedT1 Weighted T2 Weighted

28. Tumor Imaging – IP Injected Ovarian Tumor Cells
• VivoQuant was used to visualize the acquired images in 3D and
to quantify the tumor volumes.
• Various manual and semi-automatic tools are available
T2 WeightedT1 Weighted
Region Of
Interest
Color
Volume
(mm³)
Upper Tumor red 144
Mid Tumor green 6
Lower Tumor blue 9
Lower Tumor #2 cyan 7
Mid Tumor #2 magenta 64

29. Brain Imaging – Normal Brain
T2 WeightedT1 Weighted
• Various structures can be visualized within the brain
• 200µm in-plane resolution, 1mm slice thickness; 7.5-minute acquisition time

30. Sample images acquired during the live virtual
demo (Oct 1)
Understanding the Complementary Nature of
Imaging Modalities

31. U n d e r s t a n d i n g t h e C o m p l e m e n t a r y N a t u r e o f I m a g i n g M o d a l i t i e s
T1 Weighted
• Bioluminescence helps to identify where the tumors may be located, however differentiating tumors from one another, and
measuring tumor volume is better done using an anatomical imaging modality such as MRI or Ultrasound
Kidney Splenic Vein
TumorIntestine
IP Injection of SKOV3 Ovarian
Tumor Cells

32. Unders tanding the Comp lementar y Nature of Imag ing Modalities
T2 Weighted
• Bioluminescence helps to confirm viability of the tumor cells, as they express luciferase, approximate volumes may be
possible from the BLI signal; anatomical images help to confirm tumor volume - ultrasound (263mm3) or MRI (273mm3)
Orthotopic Mammary Fat Pad
Tumor (MDA-MB-231)