Publications

Cryo‑Fluorescence Tomography as a Tool for Visualizing Whole‑Body Inflammation Using Perfluorocarbon Nanoemulsion Tracers

Leach, B.I., Lister, D., Adams, S.R. et al. 

Mol Imaging Biol (2024). https://doi.org/10.1007/s11307-024-01926-w

Summary

Role of Xerra
PFC nanoemulsions were visualized using CFT within tissues of the reticuloendothelial system and inflammatory lesions, consistent with immune cell (macrophage) labeling, as previously reported in in vivo magnetic resonance and nuclear imaging studies. The CFT signals show pronounced differences among fluorescence wavelengths and tissues, presumably due to autofluorescence, differential fluorescence quenching, and scattering of incident and emitted light.
CFT Advantage
High-resolution, three-dimensional mapping of probe localization in intact animals and tissue samples, enabling unbiased validation of probe biodistribution and minimizes reliance on laborious histological methods employing discrete tissue panels, where disseminated populations of PFC-labeled cells may be overlooked. This methodology can be used to streamline the development of new generations of non-invasive, cellular-molecular imaging probes for in vivo imaging.
Preclinical Applications
Localization, Probe Biodistribution
Probes Used In Xerra™

NIR, red, and green PFC nanoemulsions

Imaging Modalities Shown
CFT

Animal Model

C57BL/6 mice bearing colon adenocarcinoma MC38 tumors; murine breast cancer 4T1 cells implanted into female Balb/c mice; Female CD-1 mice; Wild-type C57BL/6 mice; C57BL/6 mice

MRI-Based Cell Tracking of OATP-Expressing Cell Transplants by Pre-Labeling with Gd-EOB-DTPA

Bhattacharyya, T., Mallett, C.L. & Shapiro, E.M.

Mol Imaging Biol 26, 233–239 (2024). https://doi.org/10.1007/s11307-024-01904-2

Summary

Role of Xerra
Cryo-Fluorescence Tomography showed that the areas of signal enhancement in MRI overlapped with areas of indocyanine green signal, indicating that MRI signal enhancement was due to the transplanted cells.
CFT Advantage
High resolution and high sensitivity imaging that contains white-light anatomical data to help with sub organ localization
Preclinical Applications
Localization
Probes Used In Xerra™

indocyanine green

Imaging Modalities Shown
CFT, T1-Weighted MRI

Animal Model

HEK293 cells were stably transduced to express human OATP1B3 and injected into chicken hearts

Intratumoral Biosynthesis of Gold Nanoclusters by Pancreatic Cancer to Overcome Delivery Barriers to Radiosensitization

Aaron S. Schwartz-Duval, Yuri Mackeyev, Iqbal Mahmud, Philip L. Lorenzi, Mihai Gagea, Sunil Krishnan, and Konstantin V. Sokolov

ACS Nano 2024, 18, 3, 1865–1881 (2024).  https://doi.org/10.1021/acsnano.3c04260

Summary

Role of Xerra
Cryo-Fluorescence Tomography showed a fluorescence from GNCs in the Au3+-treated group that was absent in the untreated control group (Figure 6B). GNC fluorescence across multiple tumor cross sections did not fluctuate significantly, indicating a uniform distribution of GNCs in the tumor.
CFT Advantage
CFT has the ability to track fluorescently labeled gold nanoclusters with high resolution and sensitivity
Preclinical Applications
Monitor and track uptake of GNC's into tumor's
Imaging Modalities Shown
CFT, Confocal Fluorescence

Animal Model

PANC1 tumor xenografts 48 h after intratumoral injection of either 0.00 or 1.00 mM Au3+ in 20 μL of PBS.

Characterization and biodistribution of under-employed gene therapy vector AAV7

Samantha A. Yost, Emre Firlar, Justin D. Glenn, Hayley B. Carroll, Steven Foltz, April R. Giles, Jenny M. Egley, Elad Firnberg, Sungyeon Cho, Trang Nguyen, William M. Henry, Karolina J. Janczura, Joseph Bruder, Ye Liu, Olivier Danos, Subha Karumuthil-Melethil, Sanjana Pannem, Valerie Yost, Yelena Engelson, Jason T. Kaelber, Hemi Dimant, Jared B. Smith, Andrew C. Mercer


 Journal of Virology. 2023 Oct 16; 10.1128/jvi.01163-23

Summary

Role of Xerra
Cryofluorescence tomography (CFT) of whole mice was performed using an Xerra imager (Emit Imaging) 3 weeks after IV administration of AAV7 or AAV9 carrying an EGFP transgene or co-administration of AAV7.CAG.EGFP and AAV9.CAG.tdTomato (2 ×1014 GC/kg).
CFT Advantage
CFT supports the biodistribution findings in the brain, showing the presence of weak fluorescence in the brain for both AAV7 and AAV9. A surprising finding from CFT was the presence of fluorescence in facial structures, some of it highly concentrated
Preclinical Applications
Protein Expression, Biodistribution, Drug Development
Probes Used In Xerra™

EGFP, TdTomato

Imaging Modalities Shown
CFT, Cryo-electron microscopy, Immunofluorescence

Animal Model

Female C57Bl/6 mice aged 6–8 weeks (Charles River Laboratories) were acclimated for 48 hours prior to AAV administration and divided into five groups (n = 3 per group): non-treated, AAV7.CAG.EGFP, AAV9.CAG.EGFP, AAV9.CAG.TdTomato, and an equimolar mixture of AAV7.CAG.EGFP and AAV9.CAG.TdTomato.

Whole CNS 3D Cryo-Fluorescence Tomography Shows CSF Clearance along Nasal Lymphatics, Spinal Nerves, and Lumbar/Sacral Lymph Nodes

Stokes, C.; White, E.F.; Toddes, S.; Bens, N.; Kulkarni, P.; Ferris, C.F. 
 
J. Imaging 2023, 9, 45. https://doi.org/10.3390/jimaging9020045

Summary

Role of Xerra
Cryo-Fluorescence Tomography (CFT) was used to follow the movement of tracers from the ventricular system of the brain down through the meningeal lining of the spinal cord and out to the spinal lymphatic nodes.
CFT Advantage
These studies using CFT enabled followingthe distribution, localization, and clearance of Qdots over the entire CNS.
Preclinical Applications
Biodistribution, Localization, and Clearance
Probes Used In Xerra™

QdotR 605 ITKTM amino (PEG)

Imaging Modalities Shown
CFT

Animal Model

Male C57BL/J6 mice

Volumetric imaging of optically cleared and fluorescently labeled animal tissue (VIOLA) for quantifying the 3D biodistribution of nanoparticles at cellular resolution in tumor tissue

Nikolai Rakhilin, Bing Yang, Mary E. Spilker, Lisa K. Manzuk, Mary Katherine Montgomery, Eyoung Shin, Nadira Prashad, Jungyeon Hwang, Youngho Song, Frank Loganzo, Anand Giddabasappa, Sripad Ram,

Journal of Controlled Release, Volume 354, 2023, Pages 244-259, ISSN 0168-3659, https://doi.org/10.1016/j.jconrel.2022.12.060.

Summary

Role of Xerra
Imaged tumor specimens using cryo-fluorescence tomography (CFT) which measured NP biodistribution across the entire thickness of the tumor specimen.
CFT Advantage
CFT was not limited to the depth limitations of experienced with VIOLA at 1mm.
Preclinical Applications
Biodistribution
Imaging Modalities Shown
CFT, Volumetric imaging of optically cleared and fluorescently labeled animal tissue (VIOLA)

Animal Model

KPC-GEM (LSL-KrasG12D/+; LSL-Trp53R172H/+; Pdx-1-Cre) mouse was used alongside BALB/c mice (Jax #000651) for CT26 subcutaneous injection xenograft model and EMT6 mammary gland injection model.

A Genetically Encoded Magnetic Resonance Imaging Reporter Enables Sensitive Detection and Tracking of Spontaneous Metastases in Deep Tissues

Nivin N. NyströmSean W. McRaeFrancisco M. MartinezJohn J. KellyTimothy J. SchollJohn A. Ronald; A 
Cancer Res 1 March 2023; 83 (5): 673–685. https://doi.org/10.1158/0008-5472.CAN-22-2770

Summary

Role of Xerra
Cryo-Fluorescence Tomography was utilized to identify 3D detection of micrometastases of a Luc-1B3 mouse.
CFT Advantage
CFT has the ability to identify on and off target metastases in3D with high resolution and sensitivity.
Preclinical Applications
Identify and confirm location of metastases within mouse models.
Imaging Modalities Shown
CFT, BLI, T1 weighted-MRI, Histology

Animal Model

Luc-1B3 mouse burdened with lung micrometastases

Biodistribution and delivery of oligonucleotide therapeutics to the central nervous system: Advances, challenges, and future perspectives

Goto, A.Yamamoto, S., & Iwasaki, S. 
Biopharmaceutics and Drug Disposition44(1), 2647. 2023 https://doi.org/10.1002/bdd.2338

Summary

Role of Xerra
Review Article comparing imaging modalities for analyzing biodistribution
CFT Advantage
Whole animal volumetric tissue imaging technique to monitor on and off target biodistribution.
Imaging Modalities Shown
CFT, PET/SPECT, In vivo confocal microscopy, LSFM

Preclinical characterization of novel radiolabeled and fluorescent-labeled Fibroblast Activation Protein (FAP)-targeting ligands using gamma counting, SPECT imaging and Cryo-Fluorescence Tomography (CFT)

Hesterman, Novicki, White, Stokes, Cordova, Toddes, Silva, Burke, Domarkas, Wright, Archibald, Yost, Heimann, DiMagno, Hillier, Patel, Orcut, Amor, Tully, Babich, Hoppin

 

 EANM 2023

Summary

Role of Xerra
Visualize the biodistribution of the FAP imaging candidate compounds
CFT Advantage
CFT imaging provided higher resolution whole-body information, yielding insights such as the local distribution of the ligand on the surface of the bone and heterogeneity of distribution within the tumor
Preclinical Applications
Tumor Biology, Biodistribution, Drug Development
Probes Used In Xerra™

ZW800-1

Imaging Modalities Shown
SPECT/CT, CFT

Animal Model

A 1ug mass dose of either [67Cu]RTX-1363 or [ZW800-1]RTX-1341 was administered in 100 uL via tail-vain injection to U-87 MG tumor-bearing mice

STING Agonist Delivery by Tumour-Penetrating PEG-Lipid Nanodiscs Primes Robust Anticancer Immunity

Eric L. Dane, Alexis Belessiotis-Richards, Coralie Backlund , Jianing Wang, Kousuke Hidaka, Lauren E. Milling, Sachin Bhagchandani, Mariane B. Melo, Shengwei Wu, Na Li, Nathan Donahue, Kaiyuan Ni , Leyuan Ma, Masanori Okaniwa , Molly M. Stevens , Alfredo Alexander-Katz, and Darrell J. Irvine

 

Nature Materials. 21, 710-720 (2022)

Summary

Role of Xerra
High resolution, high sensitivity imaging of new agents LND-CDN carriers that are clinically viable immunostimulatory drugs to increase the response rate of immunotherapy
CFT Advantage
3D validation of histology results; full body biodistribution of agents
Preclinical Applications
Tumor Biology, Biodistribution, Drug Development
Probes Used In Xerra™

cGAMP-sulfo-Cy5

Imaging Modalities Shown
None; some histology

Animal Model

B6 mice bearing MC38 flank tumours inoculated 7d previously were intravenously administered LND or liposome both labelled with 2.0nmol sulfo-Cy5 dye per mouse via retro-orbital injection, or left untreated

High-Contrast Detection of Somatostatin Receptor Subtype2 for Fluorescence-Guided Surgery

Servando Hernandez Vargas, Solmaz AghaAmiri, Sukhen C. Ghosh, Michael P. Luciano, Luis C. Borbon, Po Hien Ear, James R. Howe, Jennifer M. Bailey-Lundberg, Gregory D. Simonek, Daniel M. Halperin, Hop S. Tran Cao, Naruhiko Ikoma, Martin J. Schnermann, and Ali Azhdarinia

 

Mol. Pharmaceutics 2022, 19, 4241−4253

Summary

Role of Xerra
Map out the distribution and signal intensity of conjugates while maintaining anatomical context in a mouse tumor model
CFT Advantage
High resolution, high sensitivity imaging of MMC(FNIRTag)-TOC and its IR800 counterpart in SSTR2+ and SSTR2− tumors
Preclinical Applications
Imaging Agent Development, Tumor Biology
Imaging Modalities Shown
Bioluminescence, Confocal Microscopy, NIRF Imaging, PET/CT

Animal Model

Athymic female J:Nu nude mice with NEC913/NEC1452 xenografts
CFT Study Details

Acquisition parameters were Ex/Em = 730 and 794 nm, respectively, and auto exposure was +3×

Microenvironment-triggered multimodal precision diagnostics

Liangliang Hao, Nazanin Rohani, Renee T. Zhao, Emilia M. Pulver, Howard Mak, Olivia J. Kelada, Henry Ko, Heather E. Fleming, Frank B. Gertler & Sangeeta N. Bhatia


Nature Materials. 20, 1440–1448 (2021).

Summary

Role of Xerra
To detect the localization of fluorescent signal in a pattern perfectly aligned with the corresponding anatomical tumors throughout the lung (imaged whole animals after systemic administration of fluorescein amidite (FAM)-labelled PRISM)
CFT Advantage
CFT was able to detect anatomical tumor location of tumors and a protease-responsive imaging sensors for malignancy (PRISM). Analysis software was used to create fly-through animations and 3D models of the whole animal models.
Preclinical Applications
Oncology, Drug Discovery
Probes Used In Xerra™

FAM fluorophore in PRISM
λEx/Em: 470nm/511/20nm.

Imaging Modalities Shown
CFT, PET/CT, Confocal Microscopy, Immunofluorescence

Animal Model

Female BALB/c mice

Emergent Fluorous Molecules and Their Uses in Molecular Imaging

Chao Wang, Stephen R. Adams, and Eric T. Ahrens

 

Accounts of Chemical Research. 2021 Aug 3;54(15):3060-3070 

Summary

Role of Xerra Visualize nanoemulsion (NE) biodistribution
Role of Xerra
Visualize nanoemulsion (NE) biodistribution
CFT Advantage
3D imaging of NE, registration of WL and FL signal, high sensitivity compared to other fluorescence imaging modalities
Preclinical Applications
Gene Therapy, Oncology, Neurology, Immunology, Drug Discovery
Probes Used In Xerra™
Cy5
λEx/Em: 673/707
Imaging Modalities Shown
CFT, 19F and 1H MRI, Bimodal PET/MRI, PET, CT, CFT, IVIS

Animal Model

Acute Inflammation Murine Model

Immunotherapy-induced antibodies to endogenous retroviral envelope glycoprotein confer tumor protection in mice

Kang BH, Momin N, Moynihan KD, Silva M, Li Y, Irvine DJ, et al.

 

(2021) PLoS ONE 16(4): e0248903. https://doi.org/10.1371/journal.pone.0248903

Summary

Role of Xerra Visualize nanoemulsion (NE) biodistribution
Role of Xerra
Visualize 1E4 effectively targeting tumor cells
CFT Advantage
Ability to detect a strong signal in the tumor, indicating that 1E4 effectively targeted tumor cells.
Preclinical Applications
Biodistribution of IE4 in a mouse bearing a B16F10 tumor.
Probes Used In Xerra™

Alexa Fluor 750 NHS Ester (Life Technologies)

Imaging Modalities Shown
CFT, 2D immunoblot

Animal Model

1E4 was labeled with Alexa Fluor 750 NHS Ester (Life Technologies) and 500 μg was intravenously injected into a naive mouse bearing a B16F10 tumor

Transcriptomic Analysis of Diffuse Intrinsic Pontine Glioma (DIPG) Identifies a Targetable ALDH-Positive Subset of Highly Tumorigenic Cancer Stem-like Cells

Rachel K. SurowiecSarah F. FerrisApril ApfelbaumCarlos EspinozaRanjit K. MehtaKaramoja MonchampVeerin R. SirihorachaiKaran BediMats LjungmanStefanie Galban

 

Mol Cancer Res 1 February 2021; 19 (2): 223–239. https://doi.org/10.1158/1541-7786.MCR-20-0464

Summary

Role of Xerra To define 2D and 3D spatiotemporal pharmacokinetics of ASO penetration in CNS structures and demonstrate their interaction with multiple molecular movement paths.

Role of Xerra

3D reconstructions of the CFT imaging demonstrated a diffuse tumor cell infiltration in the implanted brains with the most densely populated DIPGs located near the implantation site (pons) in the 555 nm excitation maximum intensity projection images and becoming more diffuse moving through the midline and frontal cortex of the brain with no clear tumor boarders.

CFT Advantage

Cell localization of tumor cell infiltration in the implanted brains.

Preclinical Applications

Neurology

Probes Used In Xerra™

mCherry

Imaging Modalities Shown
CFT, Histology, BLI

Animal Model

Murine brain from mCherry-labeled DIPG 007 ALDH+ implanted cells was removed on day 91 postimplantation

Fluorescence-guided resection of tumors in mouse models of oral cancer

Demétrio de Souza França, P., Guru, N., Roberts, S. et al.

 

Sci Rep 10, 11175 (2020). https://doi.org/10.1038/s41598-020-67958-8

Summary

Role of Xerra To define 2D and 3D spatiotemporal pharmacokinetics of ASO penetration in CNS structures and demonstrate their interaction with multiple molecular movement paths.

Role of Xerra

A mouse with oral cancer and palpable bilateral neck metastases along with a control mouse were imaged to evaluate PARPi-FL uptake

CFT Advantage

Tumor Localization

Preclinical Applications

Oncology

Probes Used In Xerra™

PARPi-FL

Imaging Modalities Shown
CFT, Epifluorescence imaging, Stereoscope imaging, confocal microscopy

Animal Model

C57BL/6J black mice, athymic nude mice

Fluorescence labeling of a NaV1.7-targeted peptide for near-infrared nerve visualization

Gonzales, J., Pirovano, G., Chow, C.Y. et al.

 

EJNMMI Res 10, 49 (2020). https://doi.org/10.1186/s13550-020-00630-4

Summary

Role of Xerra To define 2D and 3D spatiotemporal pharmacokinetics of ASO penetration in CNS structures and demonstrate their interaction with multiple molecular movement paths.
CFT Advantage

High resolution biodistribution

Preclinical Applications

CNS

Probes Used In Xerra™

Hs1a-FL

Imaging Modalities Shown
CFT, Confocal Microscopy, Epifluorescence imaging

Animal Model

Female athymic nude mice

Brain Pharmacology of Intrathecal Antisense Oligonucleotides Revealed Through Multimodal Imaging

Mazur C, Powers B, Zasadny K, Sullivan JM, Dimant H, Kamme F, Hesterman J, Matson J, Oestergaard M, Seaman M, Holt RW, Qutaish M, Polyak I, Coelho R, Gottumukkala V, Gaut CM, Berridge M, Albargothy NJ, Kelly L, Carare RO, Hoppin J, Kordasiewicz H, Swayze EE, Verma A

 

Journal of Clinical Investigation Insight. 2019 Oct 17;4(20)

Summary

Role of Xerra To define 2D and 3D spatiotemporal pharmacokinetics of ASO penetration in CNS structures and demonstrate their interaction with multiple molecular movement paths.

Role of Xerra

To define 2D and 3D spatiotemporal pharmacokinetics of ASO penetration in CNS structures and demonstrate their interaction with multiple molecular movement paths.
CFT Advantage
2D/3D imaging of ASO brain penetration, registration of WL and FL signal, and collection brain sections on glass slides for fluorescence microscopy and IHC.

Preclinical Applications

Neurology, Drug Discovery
Probes Used In Xerra™
Cy7
λEx/Em: 756/779
Imaging Modalities Shown
CFT, SPECT/CT, 18F PET, Microscopy

Animal Model

Sprague Dawley Rats

Dynamic Dual-Isotope Molecular Imaging Elucidates Principles for Optimizing Intrathecal Drug Delivery

Wolf DA, Hesterman JY, Sullivan JM, Orcutt KD, Silva MD, Lobo M, Wellman T, Hoppin J, Verma A.

 

Journal of Clinical Investigation Insight. 2016 Feb 25;1(2)

Summary

Role of Xerra To determine the effects of the neuraxial exposure of drug surrogate molecules following an IT lumbar bolus in the rat.

Role of Xerra

To determine the effects of the neuraxial exposure of drug surrogate molecules following an IT lumbar bolus in the rat.

CFT Advantage
CFT was used to follow the IT distribution of EBD, a small molecule (MW 961 Da), which binds tightly to albumin and can move from the CSF into the ISF of CNS tissue. Analysis software was used to create fly-through animations and 3D models of the rat spinal column and to estimate CSF volume by application of blue color detectors.

Preclinical Applications

Neurology, Drug Discovery
Probes Used In Xerra™
Cy5
λEx/Em: 673/707
Imaging Modalities Shown
MRI, SPECT/CT, Autoradiography, Microscopy

Animal Model

Sprague Dawley Rats

Discover More with Cryo-Fluorescence Tomography