TY - JOUR AB - Botulinum neurotoxins (BoNTs) and tetanus toxin (TeTX) are the deadliest biological substances that cause botulism and tetanus, respectively. Their astonishing potency and capacity to enter neurons and interfere with neurotransmitter release at presynaptic terminals have attracted much interest in experimental neurobiology and clinical research. Fused with reporter proteins or labelled with fluorophores, BoNTs and TeTX and their non-toxic fragments also offer remarkable opportunities to visualize cellular processes and functions in neurons and synaptic connections. This study presents the state-of-the-art optical probes derived from BoNTs and TeTX and discusses their applications in molecular and synaptic biology and neurodevelopmental research. It reviews the principles of the design and production of probes, revisits their applications with advantages and limitations and considers prospects for future improvements. The versatile characteristics of discussed probes and reporters make them an integral part of the expanding toolkit for molecular neuroimaging, promoting the discovery process in neurobiology and translational neurosciences. AU - Bijjam, R.* AU - Shorter, S.* AU - Bratt, A.M.* AU - O'Leary, V.B.* AU - Ntziachristos, V. AU - Ovsepian, S.V.* C1 - 71871 C2 - 56403 CY - One New York Plaza, Suite 4600, New York, Ny, United States TI - Neurotoxin-derived optical probes for elucidating molecular and developmental biology of neurons and synaptic connections : Toxin-derived optical probes for neuroimaging. JO - Mol. Imaging Biol. PB - Springer PY - 2024 SN - 1536-1632 ER - TY - JOUR AB - The superb specificity and potency of biological toxins targeting various ion channels and receptors are of major interest for the delivery of therapeutics to distinct cell types and subcellular compartments. Fused with reporter proteins or labelled with fluorophores and nanocomposites, animal toxins and their detoxified variants also offer expanding opportunities for visualisation of a range of molecular processes and functions in preclinical models, as well as clinical studies. This article presents state-of-the-art optical probes derived from neurotoxins targeting ion channels, with discussions of their applications in basic and translational biomedical research. It describes the design and production of probes and reviews their applications with advantages and limitations, with prospects for future improvements. Given the advances in imaging tools and expanding research areas benefiting from the use of optical probes, described here resources should assist the discovery process and facilitate high-precision interrogation and therapeutic interventions. AU - Ergen, P.H.* AU - Shorter, S.* AU - Ntziachristos, V. AU - Ovsepian, S.V.* C1 - 68085 C2 - 54563 CY - One New York Plaza, Suite 4600, New York, Ny, United States SP - 799-814 TI - Neurotoxin-derived optical probes for biological and medical imaging. JO - Mol. Imaging Biol. VL - 25 IS - 5 PB - Springer PY - 2023 SN - 1536-1632 ER - TY - JOUR AB - Intraoperative imaging (IOI) is performed to guide delineation and localization of regions of surgical interest. While oncological surgical planning predominantly utilizes x-ray computed tomography (CT), magnetic resonance imaging (MRI), and ultrasound (US), intraoperative guidance mainly remains on surgeon interpretation and pathology for confirmation. Over the past decades however, intraoperative guidance has evolved significantly with the emergence of several novel imaging technologies, including fluorescence-, Raman, photoacoustic-, and radio-guided approaches. These modalities have demonstrated the potential to further optimize precision in surgical resection and improve clinical outcomes for patients. Not only can these technologies enhance our understanding of the disease, they can also yield large imaging datasets intraoperatively that can be analyzed by deep learning approaches for more rapid and accurate pathological diagnosis. Unfortunately, many of these novel technologies are still under preclinical or early clinical evaluation. Organizations like the Intra-Operative Imaging Study Group of the European Society for Molecular Imaging (ESMI) support interdisciplinary interactions with the aim to improve technical capabilities in the field, an approach that can succeed only if scientists, engineers, and physicians work closely together with industry and regulatory bodies to resolve roadblocks to clinical translation. In this review, we provide an overview of a variety of novel IOI technologies, discuss their challenges, and present future perspectives on the enormous potential of IOI for oncological surgical navigation. AU - Alam, I.S.* AU - Steinberg, I.* AU - Vermesh, O.* AU - van den Berg, N.S.* AU - Rosenthal, E.L.* AU - van Dam, G.M.* AU - Ntziachristos, V. AU - Gambhir, S.S.* AU - Hernot, S.* AU - Rogalla, S.* C1 - 53674 C2 - 44942 CY - 233 Spring St, New York, Ny 10013 Usa SP - 705-715 TI - Emerging intraoperative imaging modalities to improve surgical precision. JO - Mol. Imaging Biol. VL - 20 IS - 5 PB - Springer PY - 2018 SN - 1536-1632 ER - TY - JOUR AB - PurposeDiabetes is associated with a deterioration of the microvasculature in brown adipose tissue (BAT) and with a decrease in its metabolic activity. Multispectral optoacoustic tomography has been recently proposed as a new tool capable of differentiating healthy and diabetic BAT by observing hemoglobin gradients and microvasculature density in cross-sectional (2D) views. We report on the use of spiral volumetric optoacoustic tomography (SVOT) for an improved characterization of BAT.ProceduresA streptozotocin-induced diabetes model and control mice were scanned with SVOT. Volumetric oxygen saturation (sO(2)) as well as total blood volume (TBV) in the subcutaneous interscapular BAT (iBAT) was quantified. Segmentation further enabled separating feeding and draining vessels from the BAT anatomical structure.ResultsScanning revealed a 46% decrease in TBV and a 25% decrease in sO(2) in the diabetic iBAT with respect to the healthy control.ConclusionsThese results suggest that SVOT may serve as an effective tool for studying the effects of diabetes on BAT. The volumetric optoacoustic imaging probe used for the SVOT scans can be operated in a handheld mode, thus potentially providing a clinical translation route for BAT-related studies with this imaging technology. AU - Ron, A. AU - Dean-Ben, X.L. AU - Reber, J. AU - Ntziachristos, V. AU - Razansky, D. C1 - 54636 C2 - 45741 CY - 233 Spring St, New York, Ny 10013 Usa SP - 620-625 TI - Characterization of brown adipose tissue in a diabetic mouse model with spiral volumetric optoacoustic tomography. JO - Mol. Imaging Biol. VL - 21 IS - 4 PB - Springer PY - 2018 SN - 1536-1632 ER - TY - JOUR AB - Atherosclerotic plaques can remain quiescent for years, but become life threatening upon rupture or disruption, initiating clot formation in the vessel lumen and causing acute myocardial infarction and ischemic stroke. Whether and how a plaque ruptures is determined by its macroscopic structure and microscopic composition. Rupture-prone plaques usually consist of a thin fibrous cap with few smooth muscle cells, a large lipid core, a dense infiltrate of inflammatory cells, and neovessels. Such lesions, termed high-risk plaques, can remain asymptomatic until the thrombotic event. Various imaging technologies currently allow visualization of morphological and biological characteristics of high-risk atherosclerotic plaques. Conventional protocols are often complex and lack specificity for high-risk plaque. Conversely, new imaging approaches are emerging which may overcome these limitations. Validation of these novel imaging techniques in preclinical models of atherosclerosis is essential for effective translational to clinical practice. Imaging the vessel wall, as well as its biological milieu in small animal models, is challenging because the vessel wall is a small structure that undergoes continuous movements imposed by the cardiac cycle as it is adjacent to circulating blood. The focus of this paper is to provide a state-of-the-art review on techniques currently available for preclinical imaging of atherosclerosis in small animal models and to discuss the advantages and limitations of each approach. AU - Vigne, J.* AU - Thackeray, J.* AU - Essers, J.* AU - Makowski, M.* AU - Varasteh, Z.* AU - Curaj, A.* AU - Karlas, A. AU - Canet-Soulas, E.* AU - Mulder, W.J.* AU - Kiessling, F.* AU - Schäfers, M.* AU - Botnar, R.M.* AU - Wildgruber, M.* AU - Hyafil, F.* C1 - 54403 C2 - 45555 CY - 233 Spring St, New York, Ny 10013 Usa SP - 869-887 TI - Current and emerging preclinical approaches for imaging-based characterization of atherosclerosis. JO - Mol. Imaging Biol. VL - 20 IS - 6 PB - Springer PY - 2018 SN - 1536-1632 ER - TY - JOUR AB - PURPOSE: A primary enabling feature of near-infrared fluorescent proteins (FPs) and fluorescent probes is the ability to visualize deeper in tissues than in the visible. The purpose of this work is to find which is the optimal visualization method that can exploit the advantages of this novel class of FPs in full-scale pre-clinical molecular imaging studies. PROCEDURES: Nude mice were stereotactically implanted with near-infrared FP expressing glioma cells to from brain tumors. The feasibility and performance metrics of FPs were compared between planar epi-illumination and trans-illumination fluorescence imaging, as well as to hybrid Fluorescence Molecular Tomography (FMT) system combined with X-ray CT and Multispectral Optoacoustic (or Photoacoustic) Tomography (MSOT). RESULTS: It is shown that deep-seated glioma brain tumors are possible to visualize both with fluorescence and optoacoustic imaging. Fluorescence imaging is straightforward and has good sensitivity; however, it lacks resolution. FMT-XCT can provide an improved rough resolution of ∼1 mm in deep tissue, while MSOT achieves 0.1 mm resolution in deep tissue and has comparable sensitivity. CONCLUSIONS: We show imaging capacity that can shift the visualization paradigm in biological discovery. The results are relevant not only to reporter gene imaging, but stand as cross-platform comparison for all methods imaging near infrared fluorescent contrast agents. AU - Deliolanis, N.C. AU - Ale, A.B.F AU - Morscher, S. AU - Burton, N.C. AU - Schaefer, K. AU - Radrich, K. AU - Razansky, D. AU - Ntziachristos, V. C1 - 30809 C2 - 33885 CY - New York SP - 652-660 TI - Deep-tissue reporter-gene imaging with fluorescence and optoacoustic tomography: A performance overview. JO - Mol. Imaging Biol. VL - 16 IS - 5 PB - Springer PY - 2014 SN - 1536-1632 ER - TY - JOUR AB - During the European Molecular Imaging Meeting (EMIM) 2013, the fluorescence-guided surgery study group held its inaugural session to discuss the clinical implementation of fluorescence-guided surgery. The general aim of this study group is to discuss and identify the steps required to successfully and safely bring intraoperative fluorescence imaging to the clinics. The focus group intends to use synergies between interested groups as a tool to address regulatory and implementation hurdles in Europe and operates within the intraoperative focus group of the World Molecular Imaging Society (WMIS) that promotes the same interests at the WMIS level. The major topics on the critical path of implementation identified within the study group were quality controls and standards for ensuring accurate imaging and the ability to compare results from different studies, regulatory affairs, and strategies to increase awareness among physicians, regulators, insurance companies, and a broader audience. These hurdles, and the possible actions discussed to overcome them, are summarized in this report. Furthermore, a number of recommendations for the future shape of the fluorescence-guided study group are discussed. A main driving conclusion remains that intraoperative imaging has great clinical potential and that many of the solutions required are best addressed with the community working together to optimally promote and accelerate the clinical implementation of fluorescence imaging towards improving surgical procedures. AU - Snoeks, T.J.* AU - van Driel, P.B.* AU - Keereweer, S.* AU - Aime, S.* AU - Brindle, K.M.* AU - van Dam, G.M.* AU - Löwik, C.W.* AU - Ntziachristos, V. AU - Vahrmeijer, A.L.* C1 - 28391 C2 - 33353 CY - New York SP - 147-151 TI - Towards a successful clinical implementation of fluorescence-guided surgery. JO - Mol. Imaging Biol. VL - 16 IS - 2 PB - Springer PY - 2014 SN - 1536-1632 ER - TY - JOUR AB - Elevated expression of cathepsins, integrins and matrix metalloproteinases (MMPs) is typically associated with atherosclerotic plaque instability. While fluorescent tagging of such molecules has been amply demonstrated, no imaging method was so far shown capable of resolving these inflammation-associated tags with high fidelity and resolution beyond microscopic depths. This study is aimed at demonstrating a new method with high potential for noninvasive clinical cardiovascular diagnostics of vulnerable plaques using high-resolution deep-tissue multispectral optoacoustic tomography (MSOT) technology METHODS AND RESULTS: MMP-sensitive activatable fluorescent probe (MMPSense 680) was applied to human carotid plaques from symptomatic patients. Atherosclerotic activity was detected by tuning MSOT wavelengths to activation-dependent absorption changes of the molecules, structurally modified in the presence of enzymes. MSOT analysis simultaneously provided morphology along with heterogeneous MMP activity with better than 200 micron resolution throughout the intact plaque tissue. The results corresponded well with epi-fluorescence images made from thin cryosections. Elevated MMP activity was further confirmed by in situ zymography, accompanied by increased macrophage influx. CONCLUSIONS: We demonstrated, for the first time to our knowledge, the ability of MSOT to provide volumetric images of activatable molecular probe distribution deep within optically diffuse tissues. High-resolution mapping of MMP activity was achieved deep in the vulnerable plaque of intact human carotid specimens. This performance directly relates to pre-clinical screening applications in animal models and to clinical decision potential as it might eventually allow for highly specific visualization and staging of plaque vulnerability thus impacting therapeutic clinical decision making. AU - Razansky, D. AU - Harlaar, N.J. AU - Hillebrands, J.L.* AU - Taruttis, A. AU - Herzog, E. AU - Zeebregts, C.J.* AU - van Dam, G.M.* AU - Ntziachristos, V. C1 - 3757 C2 - 28886 SP - 277-285 TI - Multispectral optoacoustic tomography of matrix metalloproteinase activity in vulnerable human carotid plaques. JO - Mol. Imaging Biol. VL - 14 IS - 3 PB - Springer PY - 2012 SN - 1536-1632 ER - TY - JOUR AB - PURPOSE: Real-time intraoperative near-infrared fluorescence (NIRF) imaging is a promising technique for lymphatic mapping and sentinel lymph node (SLN) detection. The purpose of this technical feasibility pilot study was to evaluate the applicability of NIRF imaging with indocyanin green (ICG) for the detection of the SLN in cervical cancer. PROCEDURES: In ten patients with early stage cervical cancer, a mixture of patent blue and ICG was injected into the cervix uteri during surgery. Real-time color and fluorescence videos and images were acquired using a custom-made multispectral fluorescence camera system. RESULTS: Real-time fluorescence lymphatic mapping was observed in vivo in six patients; a total of nine SLNs were detected, of which one (11%) contained metastases. Ex vivo fluorescence imaging revealed the remaining fluorescent signal in 11 of 197 non-sentinel LNs (5%), of which one contained metastatic tumor tissue. None of the non-fluorescent LNs contained metastases. CONCLUSIONS: We conclude that lymphatic mapping and detection of the SLN in cervical cancer using intraoperative NIRF imaging is technically feasible. However, the technique needs to be refined for full applicability in cervical cancer in terms of sensitivity and specificity. AU - Crane, L.M.A.* AU - Themelis, G. AU - Pleijhuis, R.G.* AU - Harlaar, N.J. AU - Sarantopoulos, A. AU - Arts, H.J.* AU - van der Zee, A.G.J.* AU - Vasilis, N. AU - van Dam, G.M.* C1 - 5742 C2 - 27663 SP - 1043-1049 TI - Intraoperative multispectral fluorescence imaging for the detection of the sentinel lymph node in cervical cancer: A novel concept. JO - Mol. Imaging Biol. VL - 13 PB - Academy of Molecular Imaging PY - 2011 SN - 1536-1632 ER - TY - JOUR AB - Purpose: The increasing availability of fluorescent probes for in vivo optical imaging enables the interrogation of complex biological processes in small animals serving as models for human-like tissue function and disease. However, the validation of probe bio-distribution during their development or the study of different disease models, in support of in vivo imaging studies, is not straightforward. Procedures: The imaging system developed consists of a customized multispectral planar imager that has been adapted on a commercial cryomicrotome and provides a powerful modality for ex vivo imaging of small animals. Results: The ability to capture 3D anatomical (color) and fluorescence volumetric distributions of multiple fluorescent markers in high resolution is showcased. Conclusions: Serving both as a method for accurately imaging the bio-distribution of multiple fluorescent agents inside organisms and as a modality for the validation of non-invasive methods, multispectral cryoslicing imaging offers useful insights to ex vivo optical imaging of molecular probes. AU - Sarantopoulos, A. AU - Themelis, G. AU - Ntziachristos, V. C1 - 5743 C2 - 27664 SP - 874-885 TI - Imaging the bio-distribution of fluorescent probes using multispectral epi-Illumination cryoslicing imaging. JO - Mol. Imaging Biol. VL - 13 IS - 5 PB - Academy of Molecular Imaging PY - 2011 SN - 1536-1632 ER -