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
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
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
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
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
Biopharmaceutics and Drug Disposition, 44(1), 26–47. 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 |
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 |
