Watch our hands-on demonstration of the iNSiGHT, DXA imaging system for body composition analysis. During this live demonstration, we showed the full imaging and measurement capabilities of the iNSiGHT system. These measurements included: Bone mineral density in g/cm2 Bone mineral content in g Bone area in cm2 Tissue area in cm2 Fat tissue as percentage and weight in % and g Lean tissue as percentage and weight in % and g Total weight g Additionally, specific bone length measurements can be drawn on the 2D x-ray image

1. Live Virtual
Demonstration
iNSiGHT
Preclinical DXA system for in
vivo body composition and
bone mineral density
measurements

2. Tonya Coulthard, MSc
Manager, Imaging Division
Scintica Instrumentation
Val Fajardo, PhD
Canadian Research Chair – Tissue Remodeling and Plasticity
Assistant Professor
Brock University

3. WWW.SCINTICA.COM
Topics of Discussion
• What is DXA and How Does it Work
• iNSiGHT System Overview
• Live Virtual Demonstration – *animal change normal mice*
• Acquiring Live Data, and data analysis review
• Previously acquired data – discussion with Dr. Val Fajardo
• Q&A Session
3

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What is DXA and How Does it Work
• Dual Energy X-Ray Absorptiometry
• Acquired Images
• Available Measurements
• Comparison to Other Techniques
4

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Dual Energy X-Ray Absorptiometry (DXA or DEXA)
• DXA is used to assess body composition
• Different tissues in the body have varying mass attenuation
coefficients (µn)
• Two different x-ray beams, having different energies are
generated at the source, passing through the body, hitting
the x-ray detector
• Equations are used to determine which type of tissue each
pixel on the image represents
• Bone
• Soft tissue – fat mass or lean tissue mass
Figure from Luo, Yunhua. 2017. Chapter 3 – Bone Imaging for
Osteoporosis Assessment
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Acquired Images
Images acquired using the iNSiGHT DXA system 6
X-Ray Attenuation
Image
Bone Mineral Density
Image
Color Image
Fat (Orange) and Lean (Green) Mass

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Available Measurements
7
Parameter
Unit of
Measure
Description
(available on whole animal, or from each ROI)
BMC g
Bone Mineral Contents (Bone Mass)
BMC = bone density x bone area
Fat g Fat mass
Fat Ratio % Fat Ratio = Fat/Total Mass
Lean g Fat free mass
Lean Ratio % Lean Ratio = Lean/Total Mass
Total Mass g Total Mass = Fat + Lean + Bone
BMD g/cm2 Bone Mineral Density
Bone Area cm2 Bone Area in Image
Tissue Area cm2 Tissue Area in Image
Images acquired using the iNSiGHT DXA system

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iNSiGHT System Overview
• System Components
• System Features and Benefits
• Technical Specification
• Analysis Software
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iNSiGHT: System Components
• X-Ray Benchtop Cabinet
• Self-shielded
• Integrated anesthesia
• Standard electrical connection
• Software Workstation
• Windows 10 operating system
• Offline Analysis Software
• License based for offline analysis

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iNSiGHT: System Features and Benefits
10
• Longitudinal studies
• Non-invasive data acquisition allows studying changes in body composition
over time
• Easy data acquisition
• No preparation steps other anesthesia required
• Low dose radiation
• Minimizing effects on animals during each imaging session
• Fast scan times (~25 seconds/scan)
• Allows for high throughput studies
• High-resolution images (100µm)
• Study changes in body composition on small animals, incl. mice, rats, etc.
• Wide scan area (16.5 x 25.5 cm)
• Allows for a variety of animal models to be imaged, from 10 ~ 500g

11. Hands On Demonstration
of iNSiGHT DXA System

12. iNsiGHT Small Animal DXA
Validation and Application
Val Fajardo, June 9, 2021

13. Outline
Validation
 CVs: repeated measures with and without repositioning the mouse or rat
 Application
 Low-dose lithium supplementation: an update from last webinar (May 19th,
longitudinal body composition analyses)
 DBA/2J mdx mice body composition analyses (model of Duchenne muscular
dystrophy)
 Ex vivo murine femur bone analyses (Collaboration with NASA)
13

14. Validation: CVs of female mouse
14

15. Validation: CVs of male mouse
15

16. Validation: CVs of male repositioned mouse
 Male mouse, measured 6x where the mouse was repositioned before
each scan
16
Number of values
Mean
Std. Deviation
Std. Error of Mean
Coefficient of variation
BMD (g/cm2)
6
0.06550
0.001049
0.0004282
1.601%
BMC (g)
6
0.4985
0.01540
0.006286
3.089%
Bone Area (cm2)
6
7.624
0.1570
0.06411
2.060%
Tissue Area (cm2)
6
18.37
0.3475
0.1419
1.891%
Fat (%)
6
15.60
0.1285
0.05245
0.8234%
Fat (g)
6
4.630
0.05465
0.02231
1.180%
Lean %
6
83.00
0.1595
0.06512
0.1922%
Lean(g)
6
25.04
0.1133
0.04626
0.4525%
Total Weight(g)
6
30.17
0.1593
0.06501
0.5279%

17. Validation: CVs of male rat (3-4 months old)
17
Non-repositioned CV
RepositionedCV
Number of values
Mean
Std. Deviation
Std. Error of Mean
Coefficient of variation
BMD (g/cm2)
7
0.1799
0.0003780
0.0001429
0.2101%
BMC (g)
7
5.463
0.01793
0.006777
0.3282%
Bone Area (cm2)
7
30.36
0.1376
0.05199
0.4531%
Tissue Area (cm2)
7
108.2
0.2582
0.09759
0.2386%
Fat (%)
7
11.62
0.1153
0.04360
0.9929%
Fat (g)
7
33.53
0.3294
0.1245
0.9825%
Lean %
7
86.74
0.1110
0.04197
0.1280%
Lean(g)
7
255.1
0.3781
0.1429
0.1482%
Total Weight(g)
7
294.1
0.1301
0.04916
0.04423%
Number of values
Mean
Std. Deviation
Std. Error of Mean
Coefficient of variation
BMD (g/cm2)
7
0.1769
0.002035
0.0007693
1.151%
BMC (g)
7
5.340
0.07211
0.02725
1.350%
Bone Area (cm2)
7
30.18
0.1727
0.06529
0.5723%
Tissue Area (cm2)
7
107.4
0.6508
0.2460
0.6062%
Fat (%)
7
11.61
0.1119
0.04228
0.9634%
Fat (g)
7
33.71
0.2540
0.09599
0.7535%
Lean %
7
86.79
0.1206
0.04559
0.1390%
Lean(g)
7
256.6
1.251
0.4728
0.4875%
Total Weight(g)
7
295.6
1.118
0.4226
0.3782%

18. Application: low-dose lithium and body
composition
 Project in collaboration with Dr. Rebecca
MacPherson (Associate Professor, Dept.
Health Sciences)
 Stimulate energy expenditure to combat
combat obesity
18

19. Low-dose lithium ↑ energy expenditure
 10 mg/kg/day (serum concentration of 0.02 mM) for 6-12 weeks
 Dose shown to reduce high-fat diet induced weight gain (Choi et al., 2010Vascular Pharmacology, 53: 264-272)
19
D
ark
Light
D
aily
0
1000
2000
3000
VO
2
(ml/kg
body
mass/min)
*
**
**
Control LiCl
A
8
:
0
0
1
0
:
0
0
1
2
:
0
0
1
4
:
0
0
1
6
:
0
0
1
8
:
0
0
2
0
:
0
0
2
2
:
0
0
0
:
0
0
2
:
0
0
4
:
0
0
6
:
0
0
1500
2000
2500
3000
3500
4000
VO
2
(ml/kg
body
mass/min)
Control
LiCl
Light Dark
B
Geromella et al., unpublished data

20. Does this translate to a change in body
composition?
20
Geromella et al., unpublished data
*non-invasive longitudinal studies
4
6
1
0
0
5
10
15
20
Weeks
Fat
(%)
Time, p = 0.01
Treatment, p = 0.03
4
6
1
0
60
70
80
90
100
Weeks
Lean
(%)
Time, p = 0.008
Treatment, p = 0.04
Control LiCl (0.02)

21. Energy expenditure relative to FFM
 Still elevated even after normalizing to FFM.
 What are the cellular mechanisms?
21
Geromella et al., unpublished data
D
a
r
k
L
i
g
h
t
D
a
i
l
y
0
1000
2000
3000
4000
VO
2
(ml/kg
fat-free
mass/min)
*
** *
Control LiCl
A
8
:
0
0
1
0
:
0
0
1
2
:
0
0
1
4
:
0
0
1
6
:
0
0
1
8
:
0
0
2
0
:
0
0
2
2
:
0
0
0
:
0
0
2
:
0
0
4
:
0
0
6
:
0
0
2000
2500
3000
3500
4000
4500
VO
2
(ml/kg
fat-free
mass/min)
Control
LiCl
Light Dark
B

22. Duchenne muscular dystrophy
 Duchenne muscular dystrophy (DMD): a severe
muscle wasting disease caused by the absence of
a structural protein named dystrophin.
 No cure: studying novel interventions in the mdx
mouse model is an important first step in
discovering novel therapeutic strategies.
 D2 mdx mouse is a more severe and perhaps
more clinically relevant mouse model vs the
traditionalC57 mdx mouse model.
 We will use the iNSIGHT small animal DXA to
track body composition longitudinally with
without an intervention.
22

23. D2 mdx mouse body composition
 Lower body mass vs. wild-type
 Lower lean mass vs wild-type
 Lower fat mass vs wild-type
 Hypermetabolic/catabolic?
23
Cleverdon, Silvera, et al., unpublished data

24. Muscle and bone are connected
 Often muscle wasting is associated with bone loss…
 BMD and BMC are important outcome measures for us:
 Most common treatment for DMD (i.e., corticosteroids  osteoporosis)
24
Silvera, et al., unpublished data

25. Spaceflight – if you don’t use it you lose it
Without gravity, the loss of
mechanical load leads to loss of bone
and muscle mass.
 Need effective countermeasures for
long-duration spaceflight (i.e. Mars).
 In collaboration with NASA, Dr.
WendyWard (Professor, Dept.
Kinesiology) and I are targeting
glycogen synthase kinase 3.
25

26. NASA samples - femur bones from male
mice flown in space ~30 days
 Ground control – housed on Earth in
NASA flight cages
Vivarium control – housed on Earth in
standard lab cages
 Flight – housed aboard the ISS in
NASA flight cages
 ↓ BMC ↓ BMD
 Ex vivo bone CV:
 BMC = 3.2%; BMD = 1.8%
 Is glycogen synthase kinase 3
involved in the bone and muscle
26
Silvera, et al., unpublished data

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Audience Poll

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Q&A Session
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Please enter your questions in
the Q&A section
Tonya Coulthard,
MSc
Manager, Imaging
Division,
Scintica
Val Fajardo, PhD
Canadian Research Chair –
Tissue Remodeling and Plasticity
Assistant Professor,
Brock University

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