The FMT 4000 fluorescence tomography imaging system provides the greatest utility of the FMT Systems with the ability to quantitate up to four fluorophores simultaneously. It comes with four excitation laser channels (635, 680, 750, & 790 nm).
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For research use only. Not for use in diagnostic procedures.
FMT for Fluoresence Tomography: 3 Steps to Generate Fluoresence Tomographic Data in vivo
Quick to learn. Fast to image. Easy to quantify.
STEP 1: FMT Data Generation
FMT laser-driven transillumination generates paired absorption and fluorescence data maps throughout the animal.
The anesthetized mouse is comfortably placed in PerkinElmer's portable Animal Imaging Cassette. Imaging sessions are rapid (2-3 minutes per animal), animal handling is simple and the mouse remains stable and immobilized for consistent, repetitive imaging results
STEP 2: FMT Normalization
Insert the Animal Imaging Cassette into the FMT 2500. Easily position the anesthetized animal in the Imaging Cassette within the heated imaging chamber. Gas anesthesia is maintained.
STEP 3: FMT Reconstruction
Whatever your needs, PerkinElmer has a quantitative FMT platform to provide deeper insights to your biological questions. Select the FMT 1500 for quantitative imaging in individual laboratories, in smaller facilities or for single applications. The 4-channel FMT 2500 LX is available for laboratories requiring a broader range of applications and extensive multi-modality capabilities.
|Imaging Modality||Optical Imaging|
|Optical Imaging Classification||Fluorescence Imaging|
|Product Brand Name||FMT|
|Research Areas||In Vivo Research|
The accurate quantitation of gastric emptying is crucial,for understanding the physiological, pathological and,pharmacological mechanisms underlying gastric motility,both clinically and in animal studies. Current methods,used for determining gastric emptying rates in small animals,are either terminal or involve radioactive tracers, incurring,high financial and time cost. We have developed a novel,near-infrared (NIR) fluorescent imaging agent, GastroSense™ 750, to monitor and quantitate gastric,emptying rates in murine models in vivo, non-invasively, and in real time. The imaging agent is,comprised of an acid-stable NIR fluorophore conjugated to a non-absorbable pharmacokinetic,modifier designed to be formulated in liquid or incorporated into a solid meal. Using female,8 week old BALB/c mice recipients, 0.25 nmol GastroSense 750 was orally gavaged in liquid,formulation (PBS) or fed in egg yolk, an experimental approach designed to detect changes in,gastric emptying. In vivo 3D fluorescent imaging and quantitation using the FMT®
Non-invasive preclinical imaging techniques have become important tools in biomedical research, advancing the researcher’s ability to monitor disease progression and the impact of therapeutic intervention in a variety of disease areas. With the recent advances in NIR tomographic imaging technology it has been possible to demonstrate the capabilities of fluorescence molecular tomographic (FMT®) imaging to detect and quantify fluorescent biomarkers of disease in deep tissuesites of cancer, inflammation, and infectious disease. The development of the Multispecies Imaging Module (MSIM) now allows the power of deep tissue FMT imaging to be applied to preclinical rat research models.,The MSIM allows the researcher to rapidly switch between mouse and rat imaging through the addition or removal of the mouse cassette adapter. With the adapter in place, the standard mouse imaging cassette can be inserted for mouse imaging. Removal of the adapter allows the insertion of a larger imaging cassette suitable for rats or other animals of similar size (i.e. 200-450 g body weight), with gas anesthesia delivered effectively to the animal during imaging.
Drug induced liver injury (DILI) is a major reason,for late stage termination of drug discovery,research projects, highlighting the importance,of early integration of liver safety assessment in the drug development process. A technical,approach for in vivo toxicology determination was developed using Acetaminophen (APAP),a commonly used over-the-counter analgesic and antipyretic drug, to induce acute,hepatocellular liver injury. PerkinElmer imaging technology and near infrared (NIR) labeled,Annexin V (Annexin-VivoTM 750) were used to detect and quantify and necrosis associated,with this type of liver toxicity. Both fluorescent tomographic imaging (FMT® 4000 and IVIS®,SpectrumCT), and higher throughput epifluorescence imaging (IVIS SpectrumCT), provided,excellent detection of Annexin-Vivo fluorescence in the liver. Histology and plasma alanine,transaminase (ALT) confirmed the kinetics of tissue necrosis, and more extensive liver damage,was seen but with an apparent decrease in tissue PS and plasma ALT by 48 hours, suggesting,a decline in the induction of tissue destruction. Compared to conventional plasma/serum,assays, in vivo imaging can offer fast, quantitative imaging results directly assessing the tissue,of interest.
Current means of measuring disease in preclinical models of atherosclerosis include ex vivo assessment of disease tissues post-mortem and non-invasive imaging primarily of structural and anatomic features of lesions, in vivo. A non-invasive, quantitative means of imaging known biologic profiles associated with atherosclerotic disease, in vivo, would enable a robust additional understanding and analysis of disease progression and therapeutic response in research and drug development. We report the utility of the near infrared (NIR) protease-sensing, ProSense® 750 Fluorescent Pre-clinical Imaging Agent, in combination with the FMT® 2500 Quantitative Pre-clinical Imaging System for the non-invasive quantitative measurement of atherosclerotic disease biology and related response to therapy in apolipoprotein (apo) E-deficient mice in vivo. FMT (Fluorescence Molecular Tomography) imaging measured significant increases in aortic region protease activity with a range of values that were comparable to the range seen in the ex vivo aortic arches assessed by fluorescence reflectance imaging (FRI).
An in vivo non-invasive method to determine glomerular filtration rate (GFR). Glomerular filtration rate (GFR) is the gold standard in kidney function assessment and is used to determine progression of kidney disease and drug-induced kidney toxicity. One of the most accepted ways to assess GFR is by measuring the rate of disappearance of labeled inulin from the blood; as inulin is completely filtered at the kidneys’ glomeruli (but neither secreted nor reabsorbed by the tubules), this rate of disappearance is directly proportional to GFR. We have developed a near infrared (NIR) fluorescent-labeled form of inulin (GFR-Vivo™ 680) in a spectral region providing low background and high tissue penetration (ex/em = 670/685 nm) for in vivo application. Fluorescence molecular tomographic (FMT) imaging of the heart was used to detect and quantify blood levels of GFR-Vivo 680 at multiple time points, providing the necessary data to calculate the clearance rates in individual animals. Following an intravenous bolus of NIR-Inulin in SKH-1E mice, FMT® images were acquired at 1, 5, 15, 30, and 45 minutes post-injection. Clearance rates were calculated using a two-compartment curve fitting, yielding average rates of 270 + 6 mL/min in normal mice. GFR-Vivo 680, in combination with FMT heart imaging, provides a non-invasive fluorescent imaging approach to generate consistent GFR measurements in models of kidney disease, dysfunction, and drug toxicity.
Epifluorescence (2D) imaging of superficially,implanted mouse tumor xenograft models offers,a fast and simple method for assessing tumor,progression or response to therapy. This approach,for tumor assessment requires the use of near,infrared (NIR) imaging agents specific for different,aspects of tumor biology, and this Application,Note highlights the ease and utility of multiplex,NIR fluorescence imaging to characterize the,complex biology within tumors growing in a living,mouse. IntegriSense™ 750 detects avb3 integrin,expression, BombesinRSense™ 680 detects the upregulation of bombesin receptor,(associated with tumor proliferation), Transferrin-Vivo™ 750 detects increases in,transferrin receptor (due to increased iron metabolism), MMPSense® 680 is activated,by disease-related matrix-metalloproteases (secreted by tumor cells and tumor,associated macrophages), and ProSense® 750EX detects increases in cathepsin,activity (elevated in lysosomes of tumors and inflammatory cells). OsteoSense® 680,a bone turnover imaging agent, was used as a non-tumor imaging control. These,and multiple other PerkinElmer imaging agents, can be used to characterize tumor,biology, and in this set of studies the data shows that two different tumors, HeLa,(cervical cancer) and 4T1 (breast cancer), can differ in their pattern of labeling,intensities for six distinct biological imaging agents. Such an approach is likely to,prove valuable for the full biological characterization of tumors during progression,metastasis, or response to treatment.
Fluorescent dyes have been used for many years to label cells for microscopy studies, and a variety of dyes in the visible fluorescence spectrum are available to label different cellular compartments and organelles. Efficient delivery of the fluorophore to the cell without excessively modifying surface proteins or perturbing cell function is the major biotechnological challenge. In addition, researchers have taken on the challenge of in vivo imaging, focusing on near infrared (NIR) dyes that fluoresce in a spectral region better suited for in vivo imaging due to reduced background and higher tissue penetration. Here we introduce VivoTrack™ 680, a novel NIR dye that has a long aliphatic tail to facilitate intercalation into the lipid bilayer of the cell. This agent can label a variety of cells brightly and uniformly without requiring harsh organics such as DMSO or ethanol, chemicals known to affect cell function. The labeled cells retain excellent viability and normal proliferative capacity, and significant signal is retained in dividing cells in vitro for more than 7 days. Many different cell types can be labeled for in vivo transfer or implantation, and in vivo utility is illustrated,by non-invasively tracking fluorescent primary macrophages to sites of inflammation by NIR fluorescent imaging.
Asthma is an inflammatory disease process characterized by reversible airway obstruction and airway hyperresponsiveness. This disease process is driven by activated T lymphocytes and eosinophils that are recruited to the lung upon inhalation of triggering allergens. These cells release inflammatory mediators, activate mast cells and epithelial cells and stimulate mucus secretion, ultimately leading to airway obstruction. The incidence and severity of asthma is increasing worldwide, elevating the need for clinically relevant in vivo animal models that can be used to improve the understanding of asthma biology and the development of effective therapeutics. Here we illustrate the use of PerkinElmer’s FMT® 2500LX Quantitative Pre-clinical Imaging System in combination with ProSense® 680 Fluorescent Pre-clinical Imaging Agent, a near-infrared, protease-activatable agent, for the noninvasive in vivo imaging and quantitation of pulmonary inflammation.
Complete suite of in vivo fluorescence tomography imaging solutions from PerkinElmer lets you discover more about biological targets, processes and pathways, directly in the living animal.
WMIC 2013 poster,Fluorescence Molecular Tomography (FMT) is,commonly used in small animals, most,commonly mice. A significant challenge in,imaging larger anatomies is the computation,time required to perform both the forward,model for light propagation and the matrix,inversion for computing the distribution of,fluorophores in the animal. We present a,technique for reducing the size of the problem,through the use of hybrid computations in real,space and in Fourier space, which results in a,speed-up in computation time by several,orders of magnitude.
WMIC 2013 poster,Transferrin (Tf) transports iron to all tissues, particularly those with highly metabolic cells like tumors because of their high demand for iron (for heme synthesis and as cofactors of non–heme enzymes). Given the overexpression of Tf receptors (TfR) on malignant cells, targeting TfR has been a successful approach as a strategy for pharmacological intervention in both cancer diagnosis and therapy, providing a means for selective delivery and rapid receptor-mediated internalization in tumors. While most of the imaging agents targeting TfR have involved the use of radiochemicals, in this study we generated a novel TfR-targeted agent using near infrared fluorophore labeled transferrin, Transferrin-VivoTM 750 (TFV750).
As with any functional imaging, one of the critical factors in an investigator’s ability to interpret Fluorescence Molecular Tomography (FMT) data is the reliability of the tomographic reconstructions. False negatives (missing signal) and false positives (artifacts) both hamper the accurate interpretation of data. Our study addresses the most prominent false positive artifacts with a reliable way of reducing their effect without a need for a priori assumptions about the concentration of fluorescence in the reconstruction.
A novel NIR dye for in vivo temporal tracking of labeled macrophages to sites of acute inflammation
Imaging and quantification of bombesin receptor expression in vivo - WMIC 2013 Poster,Bombesin-like peptides, including the mammalian analogs,gastrin-releasing peptide (GRP) and neuromedin B (NMB),function as growth factors in normal and neoplastic tissues.,For example, GRP receptors are found in the,gastrointestinal tract where they mediate hormone release,and epithelial cell growth. These receptors are also,overexpressed in a variety of cancers and have been used,to develop imaging tracers and as targets in radiotherapy.,As a pre-clinical alternative to using ionizing radiation, we,developed a novel near infrared (NIR) fluorescent agent,BombesinRSenseTM 680 (BRS680), designed to target and,quantify bombesin receptors in vivo.
Imaging of Cathepsin K activity in rodent models of bone turnover and soft tissue calcification
WMIC2013 Poster_,The measurement of glomerular filtration rate (GFR) is the,gold standard in kidney function assessment and is used to,determine progression of kidney disease and drug-induced,kidney toxicity. This is often assessed indirectly in,preclinical animal models either by surrogate markers, like,plasma creatinine or blood urea nitrogen. GFR is best,quantified functionally by assessing plasma clearance rates,of labeled inulin or labeled inulin analogs, but to-date this,requires blood sampling - a fairly labor intensive process.,In an effort to eliminate this procedure, we developed a,near infrared (NIR) fluorescent-labeled version of inulin,(GFR-Vivo 680 [GFR680]; ex/em = 670/685 nm) in a,spectral wavelength that offers high tissue penetration and,low background suitable for in vivo imaging.
Bacterial infection is a serious and costly clinical issue, so there is significant need for preclinical tools for assessing infection and therapeutic intervention. Rapid and non-invasive imaging technologies and targeted imaging agents would prove invaluable in diagnosing and managing bacterial infections. To this end, we developed a unique type of fluorescent agent that can quickly target bacteria, providing a fluorescent method to quantify bacterial load during infection as early as one hour post-injection. A red cationic agent (BacteriSenseTM 645) targeting the negative charge at the surface of bacteria was developed
Targeted cancer therapy aims to block key signaling pathways that are critical for tumor cell growth and survival. The blockage eventually results in cell death via apoptosis and tumor growth suppression. This strategy has proven to be safe and efficacious, and the FDA has approved several targeted therapeutics in the past decade. Encouraged by the success in clinical development, many academic and pharmaceutical researcher are in active pursuit of the improvement of next generation targeted anti-cancer drugs. As a result, many new chemical and biological entities are emerging from initial screening of in vitro, in vitro and/or in silico selection processes. From the perspective of drug development, it poses a great challenge on the next stage of in vivo validation and demands a robust, accurate, and efficient method for assessment of these candidates in living animal models.,Cancer cells are known to have abnormally increased cellular metabolism, and in the early stages of effective drug treatment, cancer cells show decreases in metabolism and proliferation. These events occur prior to overt signs of cell destruction and bioluminescence imaging technology can be used to detect early changes in tumor viability in response to targeted therapy when using luciferase-expressing tumor cell lines. However, with many therapeutic treatments, particularly high-dose treatments, these early changes in tumor viability cannot be seen by bioluminescence imaging. We applied non-invasive near infrared (NIR) fluorescence imaging to address this challenge, using PerkinElmer’s imaging agents; highly metabolic cancer cells have accompanying elevations in receptors for bombesin and transferrin on their surface that can be readily imaged using our fluorescent agents, BombesinRSenseTM 680 [BR680] and Transferrin-VivoTM 750 [TV750], respectively. In this report, we demonstrate the synergistic use of fluorescence (FLI) and bioluminescence imaging (BLI) to profile tumor metabolism and vitality, respectively, in response to a targeted anti-cancer drug, sorafenib. Sorafenib is a clinically approved tyrosine kinase inhibitor effectively blocking VEGFR, PDGFR and Raf signaling in cancer. At a higher daily dose of 120 mg/kg, the drug effectively decreases viability of HCT116-luc human colon xenograft tumors within 48 hours, coinciding with the loss of bioluminescence as well as BR680 and TV750 fluorescent signals in tumors. Interestingly, at a lower dose of 40 mg/kg, BR680 signal reduction can be observed early (within 48 hours), but no significant reduction of tumor viability by BLI and tumor volume was observed until a week later. These results suggest the potential use of metabolic fluorescent imaging agents as robust, efficient and early biomarkers for pre-clinical development of targeted cancer drugs.
Folic acid (vitamin B9) is an essential nutrient required by eukaryotic cells for survival that is acquired and internalized via folate receptors (FR) on the cell surface. In particular, it is the highly metabolic cells that upregulate FR, and many different human tumor cells, including ovarian, breast, cervical, renal, colorectal and nasopharyngeal cancer cells show significant upregulation as compared to normal tissues. As such, folic acid has been successfully exploited as a cancer specific targeting moiety for the efficient delivery of chemotherapeutic agents, drug carriers, photosensitizers and diagnostic reporters. Critical to the success of such agents is the determination of the level of FR expression for a given tumor, since weak FR-expressing cancers will not respond well to folate-targeted therapies. There is therefore a need for specific and quantitative imaging agents
Ill-posedness of diffuse fluorescence tomography,inverse problems often leads to artifacts,localized near the points where excitation light,is injected into the subject. These artifacts are,especially problematic in systems with high,levels of non-specific dye, as is often,encountered after intravenous injection of,targeting agents in small animal models.,Earlier efforts to remove these artifacts have,utilized a priori structural information to guide,the reconstruction on the location of inclusions.,In this work for fluorescence tomography, an,algorithm to reduce excitation source coupling,artifacts near the air-tissue boundary using a,least-squares method in the absence of userimposed,expectation is described. This method,is based on a technique developed by Kempner,Ripoll and Yared (Poster P422, this conference),for the algebraic reconstruction technique.
Tumor neo-vasculature, characterized by the development of abnormal, leaky and tortuous blood vessels, represents a key target for cancer imaging and therapy. Among the various recognized tools for measuring microvessel density is tomato (Lycopersicon esculentum) lectin, a single polypeptide glycoprotein that binds to sugar-containing proteins present on the endothelium. The aim of this study was to develop a near infra-red tomato lectin imaging agent to non-invasively assess tumor vasculature in vivo. Conjugation of the near infra-red fluorophore VivoTag 680XL (epsilon=210,000/M/cm; abs/em max 665/688 nm) to tomato lectin was carried out by addition of the fluorophore in a DMSO solution to lectin in aqueous sodium bicarbonate. Yields of greater than 95% were achieved, based on absorbance, with a typical loading of 2 dyes per lectin. The resulting agent, TLectinSenseTM 680 (TL680), preferentially labeled primary human umbilical vein endothelial cells. Specificity of the binding was validated by control experiments using free dye and competitive blockade with excess unlabeled tomato lectin. In vivo, non-invasive, real-time imaging and quantification of tumor neo-vasculature was performed in two models: matrigel plugs containing bFGF, VEGF and heparin injected into the flank of SKH-1 mice and nude mice bearing Lewis Lung Carcinoma tumors. Using Fluorescence Molecular Tomography (FMT®) 6 hours after TL680 (4 nmoles) intravenous injection, tumor endothelium-associated fluorescence was detected in matrigel plugs and this...
The Xenogen XGI-8 Gas Anesthesia System is designed to work with the IVIS Imaging System, a technology from Xenogen that allows researchers to use r eal-time in vivo imaging to monitor and record cellular and genetic activity within a living organism.