TECHNICAL BRIEF

3D optical andmicroCT data –The power ofmultimodality
imaging.

Introduction
The IVIS® Spectrum 2 in vivo imaging system provides highly
sensitive 2D and 3D molecular imaging using bioluminescenceand fluorescence reporter genes and/or probes; advanced
2D fluorescence functionality includes transillumination andspectral unmixing capabilities.
The Quantum GX3 microCT imaging system supports in vivo
anatomical, physiological, functional and metabolic imaging
via fast, low dose gated scans in a wide variety of animalmodels. Subvolume reconstructions allow users to redo thereconstruction of static images at higher resolution without theneed to rescan the subject.

Key benefits• Easily obtain molecular,functional, and anatomical datafrom a single subject• Seamlessly co-register 3D opticaland microCT data with justone click

The rationale behind the combination of the IVIS Spectrum 2
optical imaging system and Quantum GX3 microCT scanners
lies in the ability to combine 3D anatomical, functional andmolecular imaging modalities via automatic coregistrationof datasets. Coregistration enables accurate anatomical
localization of the optical signal and depth-corrected
quantification of signal intensity. In addition, each system canbe used alone as either a single high-throughput IVIS opticalimaging system or a high-resolution microCT scanner.

3D optical and microCT data – The power of multimodality imaging.

Figure 1: Schematic illustration of a multimodality kit to combine 3D optical and microCT data.
Table 1: Key Advantages of combining 3D optical and MicroCT imaging.
3D Optical only

MicroCT only

3D Optical + MicroCT

Non-invasive imaging technologythat affords researchers the abilityto longitudinally assess biologicalprocesses, disease progression andtherapeutic efficacy in the same
animal, thus reducing the numberof animals needed while generating
more robust data.

The Quantum GX3 microCT system provides high-resolution (2.3um) microCT scanning of specimens and live animals up to 5 kg
in weight. Multiple fields of view (18, 36, 72, and 86 mm) allow
for multispecies imaging, including mouse, rat, ferret, guinea
pig, rabbit, and others. With a 163 mm bore diameter and ascannable length of 385 mm, the Quantum GX3 can imageanimals as large as rabbits (<5 kg).

Co-registration enables researchers
to harness the capabilities of each
imaging modality to elucidate
molecular and functional informationfrom optical readouts and anatomical
context from microCT data, whichallows for more comprehensive
investigation of animal models.

Broadly applicable imaging
technique that has been used to
explore disease models including,but not limited to, oncology,
infectious diseases, neurobiology,
vasculature, diabetes, gene therapy,
tissue engineering, inflammationand immunology.

The Quantum GX3 has a highly sensitive, high‑resolution CMOS
flat panel X-ray detector, which allows for faster scan times withfewer X-rays compared to most other CT systems. PreclinicalmicroCT is often excluded from use in longitudinal studies due
to high radiation doses which could impact the biology of theanimal. The CMOS X-ray flat panel detector enables fast scantimes allowing for high signal-to-noise ratios and repeated CT
scans of animals with low X-ray exposure.

Co-registration of 3D optical sources
with microCT provides anatomical
context and allows for pinpoint
localization of optical signal at depth,
a powerful feature for virtually every
end-user application.

Provides molecular and functional
insights into preclinical diseasemodels.

The Quantum GX3 microCT system features an advanced
subvolume reconstruction workflow, creating high-resolution
images from original in vivo or ex vivo CT scans.. Thisfunctionality improves overall experimental workflows, enablinghigh-resolution images to be generated without the need torescan animals, reducing time and X-ray exposure to the animal.

Multimodal imaging with optical andmicroCT gives users the ability toimage multiple readouts in the same
animal in order to answer more
complex biological questions withfewer animals.

3D tomographic algorithms correct
for tissue absorption and scatteringof light and enable the mostaccurate location and quantificationof optical sources at depth,
and calibration curves translate
radiance units into pmol of dye orcell number.

The Quantum GX3 utilizes proprietary image‑based algorithms(instead of external monitoring) to reliably synchronize and,retrospectively, gate the microCT data to produce motion-freeimages; with a fast scan time and low radiation dose (4 min, 131mGy at 70 kV). This workflow is ideal for in vivo cardiac andrespiratory applications, while maintaining the Quantum GX3’s
advantageous high-throughput scanning capabilities.

Lung or cardiac gated CT scans canbe combined with 3D optical images
to more accurately identify opticalsignal and measure functional organreadouts.

3D reconstructions can elucidate
pinpoint localization of colocalizedprobes at depth.

The Quantum GX3 microCT control panel was designed withthe ease of use for the end user in mind. Thanks to constantinnovation and factoring in customer feedback, every iterationof the software is engineered to ensure researcher success inCT imaging studies.

The multimodality mouse imaging
shuttle has built-in fiducial markers toseamlessly co-register microCT with
3D optical data acquired on Revvity’s
IVIS Spectrum optical imaging
system with one-click automatedco-registration.

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3D optical and microCT data – The power of multimodality imaging.

AB

Figure 2: Study using microCT with (A) and without (B) coregistered 3D optical imaging.

A. Mice were injected with 1 million IVISbrite™ 4T1-rLuc
tumor cells directly into lungs 2 weeks prior andWhy combine 3D optical with 3D microCT?

IVISense™ Integrin Receptor 750 fluorescent probe 24 hThis study demonstrates how harnessing the power ofprior to imaging on the IVIS system. 3D bioluminescence

two standalone imaging modalities can provide a better

imaging (BLI) data demonstrates significant tumor burden

understanding of disease biology. In this case, 3D opticalin the lung, while 3D fluorescence imaging (FLI) detected

readouts (BLI and FLI) allow researchers to gain molecularan IVISense Integrin Receptor 750 in the lung cavity,and functional information from disease models, andspecifically in areas of heavy tumor burden as indicatedco-registration of 3D optical reconstructions with microCT

by colocalized signal. This data suggests αvβ3 Integrin

data adds anatomical context to answer more complex

upregulation in tumor cells, indicating growth, viability

biological questions.

and angiogenesis.
B.	Mice were then imaged using image-based retrospective
lung gated imaging on the Quantum microCT, and results

About Revvity In Vivo imaging

demonstrate that diseased lungs showed a decrease in

Whatever approach you choose, having a supplier that has

normal healthy lung tissue.

the resources and expertise to partner with you on your	Collectively, these results highlight the benefits ofmultimodality imaging: here, 3D optical imaging provides
molecular and functional data, such as the precise
number of cells in the diseased lungs and upregulation ofcertain biological events. Adding longitudinal microCT

in vivo imaging can help you avoid potential pitfalls, and get
the quality results you need, faster. Revvity offers a range ofimaging technologies to address the breadth of applications
that require a non-invasive, sensitive and/or high-resolution
imaging approach.

imaging shows the associated reduction in normal
healthy lung tissue in the diseased animal.

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