Longitudinale imagerie est utilisée dans les études pré-cliniques de suivre la progression d&#39;une maladie ou de mesurer l&#39;effet d&#39;une thérapeutique. En oncologie, les méthodes optiques de fournir des outils rigoureux pour surveiller la croissance tumorale et de livrer quantification précise de la croissance cellulaire ou l&#39;expression des gènes à chaque point du temps dans une telle étude. Les changements anatomiques peuvent être mesurés en utilisant une technique à haute résolution comme microCT, mais pour l&#39;imagerie longitudinale une faible dose de rayons X doit être utilisé pour éviter les artefacts biologiques. Les images optiques et microCT peuvent être co-enregistré pour fournir une combinaison de données fonctionnelles et anatomiques veillant à ce que le maximum d&#39;informations sont extraites des modèles animaux. 1. Préparation de cellules <ol><li> Caliper propose une gamme de luciférase exprimant des lignées cellulaires de cancer pour la recherche pré-clinique dans des modèles murins. </li><li> MDA-MB-231-luc-D2H2LN est un être humain ligne de cellules tumorales mammaires exprimant le gène de la luciférase, qui peut être utilisé comme un indicateur optique de tumorigenèse in vivo. Cette lignée cellulaire est créé à partir d&#39;une métastase ganglionnaire spontanée qui provient d&#39;une tumeur D3H1 pad mammaires graisse et est connu pour former des métastases agressive. </li><li> Les cellules sont fournies en tant que culture exempts d&#39;agents pathogènes congelés qui pousse facilement dans les supports standard sans avoir besoin de marqueurs de sélection. </li><li> Pour vérifier l&#39;activité luciférase avant injection dans l&#39;animal une fiole de 90% de confluence est récoltée par trypsinisation. L&#39;activité luciférase est mesurée par dosage 50.000 cellules dans une plaque de microtitration et performants de dilution en série. </li></ol> 2. Injection intracardiaque de cellules chez les animaux <ol><li> Avant l&#39;injection, les animaux sont anesthésiés à l&#39;aide de 3% d&#39;isoflurane. </li><li> 1 à 3.000.000 cellules sont injectées dans le volume 50ul dans le ventricule gauche. Suspension de cellules contient 150ug/ml D-luciférine de valider la technique d&#39;injection. </li><li> Pour confirmer une souris correcte injection intracardiaque sont imagés dans le système IVIS. Les souris qui montrent le signal à travers le corps de BLI ont été injectés correctement. Si le signal est localisé les cellules ont été injectées pas correctement dans le cœur. </li></ol> 3. BLI imagerie pour surveiller les métastases <ol><li> La navette imagerie supporte la souris pour saisir les données dans les deux spectre IVIS et le système QuantumFX microCT. </li><li> La navette permet de maintenir l&#39;animal dans la même position pour l&#39;imagerie et intègre les signaux repères afin d&#39;aider à la co-registration automatique. La chambre est reliée à l&#39;évaporateur isofluorance pour maintenir l&#39;anesthésie. </li><li> Imagerie assistant dans le logiciel Image Vivre 4.1 peut être utilisé pour sélectionner automatiquement le temps d&#39;exposition, f-stop et binning et d&#39;assurer la collecte de données quantitatives. </li><li> Plusieurs images peuvent être acquises et comparées dans les études longitudinales couvrant secondes ou quelques mois selon la nature de l&#39;expérience. </li><li> Après l&#39;injection de cellules Jour 6, BLI imagerie avec IVIS spectre montre des métastases aux os dans sept des neuf souris. </li></ol> 4. Suivi longitudinal des lésions ostéolytiques à l&#39;aide de micro CT <ol><li> Les souris qui a montré des métastases osseuses clairement visible à l&#39;aide de BLI ont été imagées dans le système Quantum FX micro CT. </li><li> Le Quantum FX micro CT système délivre des images de haute qualité à une dose de rayons X suffisamment bas pour permettre aux vrais longitudinale micro CT dans les études précliniques. </li><li> La dégradation des os dans les deux jambes peuvent être surveillés à l&#39;aide longitudinalement ce système en moins d&#39;une minute. </li><li> Les souris sont anesthésiées avec 3% d&#39;isoflurane et peuvent être transférés directement à partir du système Spectrum IVIS. </li><li> Deux images de 60mm sont cousues ensemble pour capturer le point de vue animal entier. </li><li> Un champ de vision de 30mm est idéale pour l&#39;imagerie du genou en haute résolution. </li><li> Un seul balayage avec une résolution suffisamment élevée pour mesurer et quantifier les métastases osseuses ne prend que 18 secondes et délivre une dose de 13mGy seulement. </li><li> Les souris ont été imagées jusqu&#39;à 8 fois en 36 jours avec le système de micro-CT sans affecter la croissance de la tumeur ou la santé de l&#39;animal. </li><li> Le micro-imagerie CT a montré que l&#39;os leisions étaient visibles sur Jour 10 et a progressé très rapidement, ce qui correspond à l&#39;augmentation du signal de BLI. </li></ol> 5. 3D co-enregistrement et d&#39;analyse de BLI et UCT <ol><li> Une fois les images ont été capturées sur les deux le spectre IVIS et Quantum FX micro CT données du système peuvent être co-enregistrée dans une reconstruction 3D. </li><li> MicroCT images sont exportés sous forme de fichiers DICOM et chargé dans l&#39;image vivante 4,1 outil 3D multi modalité. </li><li> Dans le logiciel d&#39;image habitable exécuter le programme d&#39;DLit trois dimensions reconstruire le signal optique. </li><li> L&#39;outil de repères dans le logiciel est utilisé pour la co-enregistrer le signal optique BLI à l&#39;image des micro CT. </li><li> Recalées ensembles de données peuvent être analysées quantitativement pour surveiller plusieurs paramètres lors de progression de la maladie ici, nous assurons le suivi de croissance de la tumeur métastatique avec le développement de la lésion ostéolytique dans l&#39;os. </li> &lt;/ Ol&gt;

<ol>
	<li>Jenkins, D. E., Hornig, Y. S., Oei, Y., Dusich, J., Purchio, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioluminescent+Human+Breast+Cancer+Cell+Lines+that+Permit+Rapid+and+Sensitive+in+vivo+Detection+of+Mammary+Tumors+and+Multiple+Metastases+in+Immune+Deficient+Mice.">Bioluminescent Human Breast Cancer Cell Lines that Permit Rapid and Sensitive in vivo Detection of Mammary Tumors and Multiple Metastases in Immune Deficient Mice.</a> Breast Cancer Research. 7, R444-R454 (2005).</li><li>Cowey, S., Szafran, A. A., Kappes, J., Zinn, K. R., Siegal, G. P., Desmond, R. A., Kim, H., Evans, L., Hardy, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Breast+Cancer+Metastasis+to+Bone:+Evaluation+of+Bioluminescent+Imaging+and+MicroSPECT/CT+for+Detecting+Bone+Metastasis+in+Immunodeficient+Mice.">Breast Cancer Metastasis to Bone: Evaluation of Bioluminescent Imaging and MicroSPECT/CT for Detecting Bone Metastasis in Immunodeficient Mice.</a> Clinical and Experimental Metastasis. 24, 389-401 (2007).</li><li>Labrinidis, A., Hay, S., Liapis, V., Ponomarev, V., Findlay, D. M., Evdokiou, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zoledronic+Acid+Inhibits+Both+the+Osteolytic+and+Osteoblastic+Components+of+Osteosarcoma+Lesions+in+a+Mouse+Model.">Zoledronic Acid Inhibits Both the Osteolytic and Osteoblastic Components of Osteosarcoma Lesions in a Mouse Model.</a> Clinical Cancer Research. 15, (2009).</li><li>Kaijzel, E. L., Pluijm, G. v. a. n. d. e. r., Lowik, C. W. G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Whole-Body+Optical+Imaging+in+Animal+Models+to+Assess+Cancer+Development+and+Progression.">Whole-Body Optical Imaging in Animal Models to Assess Cancer Development and Progression.</a> Clinical Cancer Research. 13, 3490-3497 (2007).</li><li>Minn, A. J., Kang, Y., Serganova, I., Gupta, G. P., Giri, D. D., Doubrovin, M., Ponomarev, V., Gerald, W. L., Blasberg, R., Massague, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinct+Organ-Specific+Metastatic+Potential+of+Individual+Breast+Cancer+Cells+and+Primary+Tumors.">Distinct Organ-Specific Metastatic Potential of Individual Breast Cancer Cells and Primary Tumors.</a> The Journal of Clinical Investigation. 115, (2005).</li><li>Xu, H., Christensen, A., Kuo, C., Nilson, D., Lim, E., Whalen, J., Kim, J. B., Zhang, N., Singh, R., Oldfield, S., Troy, T., Rice, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Co-registration+of+in+vivo+Optical+Tomography+and+Micro-CT+for+Longitudinal+Studies.">Co-registration of in vivo Optical Tomography and Micro-CT for Longitudinal Studies.</a> Word Molecular Imaging Congress. , (2010).</li></ol>

Les auteurs sont des employés de Caliper Life Sciences qui font l&#39;instrument présenté dans cet article.

Imagerie du petit animal avec des microCT a été appliquée à la surveillance des lésions ostéolytiques dans des conditions pathologiques, tels que le développement des tumeurs (Cowey et al 2007;.. Labrinidis et al, 2009). Comme l&#39;imagerie par bioluminescence de la luciférase tumeurs étiquetées a été largement adopté pré-clinique, la co-registration des images de bioluminescence et microCT fourni une meilleure définition de la localisation anatomique des signaux de bioluminescence (Kaijzel et al. 2007). L&#39;étude précédente avec une ligne luciférase maternel étiquetés cellules tumorales MDA-MB-231-luc-D3H2LN montré des métastases aux os et le développement des lésions ostéolytiques suivants l&#39;injection intra-cardiaque (Jenkins et al 2005;. Minn et al, 2005). Dans cette étude, nous avons exploré une étude longitudinale des métastases de la tumeur et l&#39;activité ostéolytique avec combinatoire optique et faible dose de rayonnement d&#39;imagerie microCT avec le modèle de métastases MDA-MB-231-luc-D3H2LN systémique. Nous avons effectué imagerie séquentielle avec IVIS spectre et Quantum FX systèmes microCT et la bioluminescence atteint et CT co-enregistrement avec notre image habitable nouvellement développé 4.1 du logiciel (Heng et al., 2010). L&#39;imagerie par bioluminescence avec IVIS fournit non invasive de détection des cellules tumorales avec une sensibilité élevée, permettant à la tumeur pour être détecté à un stade précoce. Imagerie avec Quantum FX permet la reconstruction 3D précise des lésions osseuses et des informations détaillées sur l&#39;anatomie. Quantum FX système offre peu d&#39;imagerie dose de rayonnement, ce qui permet un suivi longitudinal de la progression de la maladie. Corrélation des lésions osseuses et des métastases tumorales a été dépeint dans l&#39;image co-enregistrées, et l&#39;avantage de suivre le développement des lésions de la multimodalité longitudinalement a été clairement démontrée.

monitoring tumor metastases and osteolytic lesions with bioluminescence and micro ct imaging

Après l&#39;accouchement intracardiaque de cellules MDA-MB-231-luc-D3H2LN cellules souris nu / nu, les métastases systémiques développés chez les animaux injectés. L&#39;imagerie par bioluminescence à l&#39;aide du spectre IVIS a été utilisé pour surveiller la distribution et le développement des cellules tumorales en suivant la procédure de livraison, y compris la reconstruction DLit pour mesurer le signal de la tumeur et sa localisation. Développement de lésions métastatiques des tissus osseux déclenche une activité ostéolytique et des lésions au tibia et du fémur ont été évaluées en utilisant des micro CT longitudinalement. Imaging a été réalisée en utilisant un Quantum FX micro CT système d&#39;imagerie rapide et à faible dose de rayons X. La faible dose de radiation permet des sessions multiples pour l&#39;imagerie être réalisée avec un cumulatif de dosage de rayons X bien en dessous DL50. Un dispositif de navettes de la souris a été utilisé pour l&#39;imagerie séquentielle image de la souris avec les deux IVIS spectre et Quantum FX réaliser un positionnement précis des animaux dans les deux la bioluminescence et les images CT. L&#39;optique et CT ensembles de données ont été co-enregistré en 3-dimentions utilisant l&#39;image vivante 4.1 du logiciel. Cette approche multi-mode permet une surveillance étroite de la croissance tumorale et le développement simultané de l&#39;activité ostéolytique.

Watch this Scientific Journal Video about Monitoring Tumor Metastases and Osteolytic Lesions with Bioluminescence and Micro CT Imaging at JoVE.com

Monitoring Tumor Metastases and Osteolytic Lesions with Bioluminescence and Micro CT Imaging

Un modèle expérimental chez la souris des métastases osseuses a été établie suivant la livraison de la luciférase intracardiaque de cellules exprimant la tumeur mammaire. Développement de la tumeur et a entraîné des lésions ostéolytiques ont été suivis longitudinalement avec la bioluminescence et la micro imagerie CT.

Surveillance des métastases tumorales et les lésions ostéolytiques avec bioluminescence et Micro CT Imaging

Following intracardiac delivery of MDA-MB-231-luc-D3H2LN cells to Nu/Nu mice, systemic metastases developed in the injected animals. Bioluminescence ...

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Research

JoVE Journal

Medicine

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Vidéo: Monitoring Tumor Metastases and Osteolytic Lesions with Bioluminescence and Micro CT Imaging

In vivo Bioluminescence Imaging of Tumor Hypoxia Dynamics of Breast Cancer Brain Metastasis in a Mouse Model

It is well recognized that tumor hypoxia plays an important role in promoting malignant progression and affecting therapeutic response negatively. There is little knowledge about in situ, in vivo, tumor hypoxia during intracranial development of malignant brain tumors because of lack of efficient means to monitor it in these deep-seated orthotopic tumors. Bioluminescence imaging (BLI), based on the detection of light emitted by living cells expressing a luciferase gene, has been rapidly adopted for cancer research, in particular, to evaluate tumor growth or tumor size changes in response to treatment in preclinical animal studies. Moreover, by expressing a reporter gene under the control of a promoter sequence, the specific gene expression can be monitored non-invasively by BLI. Under hypoxic stress, signaling responses are mediated mainly via the hypoxia inducible factor-1&alpha; (HIF-1&alpha;) to drive transcription of various genes. Therefore, we have used a HIF-1&alpha; reporter construct, 5HRE-ODD-luc, stably transfected into human breast cancer MDA-MB231 cells (MDA-MB231/5HRE-ODD-luc). In vitro HIF-1&alpha; bioluminescence assay is performed by incubating the transfected cells in a hypoxic chamber (0.1% O2) for 24 hr before BLI, while the cells in normoxia (21% O2) serve as a control. Significantly higher photon flux observed for the cells under hypoxia suggests an increased HIF-1&alpha; binding to its promoter (HRE elements), as compared to those in normoxia. Cells are injected directly into the mouse brain to establish a breast cancer brain metastasis model. In vivo bioluminescence imaging of tumor hypoxia dynamics is initiated 2 wks after implantation and repeated once a week. BLI reveals increasing light signals from the brain as the tumor progresses, indicating increased intracranial tumor hypoxia. Histological and immunohistochemical studies are used to confirm the in vivo imaging results. Here, we will introduce approaches of in vitro HIF-1&alpha; bioluminescence assay, surgical establishment of a breast cancer brain metastasis in a nude mouse and application of in vivo bioluminescence imaging to monitor intracranial tumor hypoxia.

In vivo Bioluminescence Imaging of Tumor Hypoxia Dynamics of Breast Cancer Brain Metastasis in a Mouse Model

Bioluminescence imaging of hypoxia inducible factor-1&alpha; activity is applied to monitor intracranial tumor hypoxia development in a breast cancer brain metastasis mouse model.

Dans l&#39;imagerie de bioluminescence in vivo de la tumeur Dynamics hypoxie des métastases cérébrales du cancer du sein dans un modèle murin

It is well recognized that tumor hypoxia plays an important role in promoting malignant progression and affecting therapeutic response negatively. There ...

Recherche

Médecine

Osmotic Drug Delivery to Ischemic Hindlimbs and Perfusion of Vasculature with Microfil for Micro-Computed Tomography Imaging

Preclinical research in animal models of peripheral arterial disease plays a vital role in testing the efficacy of therapeutic agents designed to stimulate microcirculation. The choice of delivery method for these agents is important because the route of administration profoundly affects the bioactivity and efficacy of these agents1,2. In this article, we demonstrate how to locally administer a substance in ischemic hindlimbs by using a catheterized osmotic pump. This pump can deliver a fixed volume of aqueous solution continuously for an allotted period of time. We also present our mouse model of unilateral hindlimb ischemia induced by ligation of the common femoral artery proximal to the origin of profunda femoris and epigastrica arteries in the left hindlimb. Lastly, we describe the in vivo cannulation and ligation of the infrarenal abdominal aorta and perfusion of the hindlimb vasculature with Microfil, a silicone radiopaque casting agent. Microfil can perfuse and fill the entire vascular bed (arterial and venous), and because we have ligated the major vascular conduit for exit, the agent can be retained in the vasculature for future ex vivo imaging with the use of small specimen micro-CT3.

We show here the in vivo insertion of an osmotic pump for constant local drug delivery and the creation of hindlimb ischemia in a mouse model. Moreover, the hindlimb vasculature is perfused with Microfil, a silicone radiopaque agent, to prepare for micro-computed tomography (micro-CT) imaging.

Osmotique Drug Delivery à hindlimbs ischémiques et la perfusion de la vascularisation avec Microfil pour Micro-en tomodensitométrie

Preclinical research in animal models of peripheral arterial disease plays a vital role in testing the efficacy of therapeutic agents designed to ...

Bioluminescence Imaging of an Immunocompetent Animal Model for Glioblastoma

In contrast to commonly reported human glioma xenograft animal models, GL261 murine glioma xenografts recapitulate nearly all relevant clinical and histopathologic features of the human disease. When GL261 cells are implanted intracranially in syngeneic C57BL/6 mice, the model has the added advantage of maintaining an intact immune microenvironment. Stable expression of luciferase in GL261 cells allows non-invasive cost effective bioluminescence monitoring of intracranial tumor growth. We have recently demonstrated that luciferase expression in GL261 cells does not affect the tumor growth properties, tumor cell immunomodulatory cytokine expression, infiltration of immune cells into the tumor, or overall survival of animals bearing the intracranial tumor. Therefore, it appears that the GL261 luciferase glioma model can be useful in the study of novel chemotherapeutic and immunotherapeutic modalities. Here we report the technique for generating stable luciferase expression in GL261 cells and how to study the in vitro and in vivo growth of the tumor cells by bioluminescence imaging.

GL261 glioma cells provide a useful immunocompetent animal model of glioblastoma. The goals of this protocol are to demonstrate proper techniques for monitoring intracranial tumor growth using in vivo bioluminescence imaging, and to verify the utility of luciferase-modified GL261 cells for studying tumor immunology and immunotherapeutic approaches for treating glioblastoma.

Bioluminescence imagerie d&#39;un modèle animal pour immunocompétentes glioblastome

In contrast to commonly reported human glioma xenograft animal models, GL261 murine glioma xenografts recapitulate nearly all relevant clinical and ...

Bioluminescence-Based Tumor Quantification Method for Monitoring Tumor Progression and Treatment Effects in Mouse Lymphoma Models

Although bioluminescence imaging (BLI) shows promise for monitoring tumor burden in animal models of cancer, these analyses remain mostly qualitative. Here we describe a method for bioluminescence imaging to obtain a semi-quantitative analysis of tumor burden and treatment response. This method is based on the calculation of a luminoscore, a value that allows comparisons of two animals from the same or different experiments. Current BLI instruments enable the calculation of this luminoscore, which relies mainly on the acquisition conditions (back and front acquisitions) and the drawing of the region of interest (manual markup around the mouse). Using two previously described mouse lymphoma models based on cell engraftment, we show that the luminoscore method can serve as a noninvasive way to verify successful tumor cell inoculation, monitor tumor burden, and evaluate the effects of in situ cancer treatment (CpG-DNA). Finally, we show that this method suits different experimental designs. We suggest that this method be used for early estimates of treatment response in preclinical small-animal studies.

Bioluminescence imaging is a well-known tool for localizing tumors and metastases, but quantification of these images often requires complex calculations and particular instruments. We describe the easy-to-use luminoscore method, based on precise acquisition conditions, requiring no calculations, and enabling tumor burden and treatment response to be monitored in mouse models.

Méthode tumeur Quantification bioluminescence-Basé pour Progression Surveillance de la tumeur et les effets de traitement dans les modèles de souris lymphome

Although bioluminescence imaging (BLI) shows promise for monitoring tumor burden in animal models of cancer, these analyses remain mostly qualitative. ...

Combined Near-infrared Fluorescent Imaging and Micro-computed Tomography for Directly Visualizing Cerebral Thromboemboli

Direct thrombus imaging visualizes the root cause of thromboembolic infarction. Being able to image thrombus directly allows far better investigation of stroke than relying on indirect measurements, and will be a potent and robust vascular research tool. We use an optical imaging approach that labels thrombi with a molecular imaging thrombus marker&#160;&#8212; a Cy5.5 near-infrared fluorescent (NIRF) probe that is covalently linked to the fibrin strands of the thrombus by the fibrin-crosslinking enzymatic action of activated coagulation factor XIIIa during the process of clot maturation. A micro-computed tomography (microCT)-based approach uses thrombus-seeking gold nanoparticles (AuNPs) functionalized to target the major component of the clot: fibrin. This paper describes a detailed protocol for the combined in vivo microCT and ex vivo NIRF imaging of thromboemboli in a mouse model of embolic stroke. We show that in vivo microCT and fibrin-targeted glycol-chitosan AuNPs (fib-GC-AuNPs) can be used for visualizing both in situ thrombi and cerebral embolic thrombi. We also describe the use of in vivo microCT-based direct thrombus imaging to serially monitor the therapeutic effects of tissue plasminogen activator-mediated thrombolysis. After the last imaging session, we demonstrate by ex vivo NIRF imaging the extent and the distribution of residual thromboemboli in the brain. Finally, we describe quantitative image analyses of microCT and NIRF imaging data. The combined technique of direct thrombus imaging allows two independent methods of thrombus visualization to be compared: the area of thrombus-related fluorescent signal on ex vivo NIRF imaging vs. the volume of hyperdense microCT thrombi in vivo.

This protocol describes the application of combined near-infrared fluorescent (NIRF) imaging and micro-computed tomography (microCT) for visualizing cerebral thromboemboli. This technique allows the quantification of thrombus burden and evolution. The NIRF imaging technique visualizes fluorescently labeled thrombus in excised brain, while the microCT technique visualizes thrombus inside living animals using gold-nanoparticles.

Combiné proche infrarouge imagerie fluorescente et la tomographie par Micro-calculé pour Directement Visualizing cérébrale thromboembolies

Direct thrombus imaging visualizes the root cause of thromboembolic infarction. Being able to image thrombus directly allows far better investigation of ...

Determining Glucose Metabolism Kinetics Using 18F-FDG Micro-PET/CT

This paper describes the use of 18F-FDG and micro-PET/CT imaging to determine in vivo glucose metabolism kinetics in mice (and is transferable to rats). Impaired uptake and metabolism of glucose in multiple organ systems due to insulin resistance is a hallmark of type 2 diabetes. The ability of this technique to extract an image-derived input function from the vena cava using an iterative deconvolution method eliminates the requirement of the collection of arterial blood samples. Fitting of tissue and vena cava time activity curves to a two-tissue, three compartment model permits the estimation of kinetic micro-parameters related to the 18F-FDG uptake from the plasma to the intracellular space, the rate of transport from intracellular space to plasma and the rate of 18F-FDG phosphorylation. This methodology allows for multiple measures of glucose uptake and metabolism kinetics in the context of longitudinal studies and also provides insights into the efficacy of therapeutic interventions.

Determining Glucose Metabolism Kinetics Using 18F-FDG Micro-PET/CT

This study describes a protocol that uses 18F-FDG and positron emission tomography/computed tomography (PET/CT) imaging, together with kinetic modelling, to quantify the in vivo, real-time uptake of 18F-FDG into tissues.

La détermination de métabolisme du glucose Kinetics L&#39;utilisation<sup&gt; 18</sup&gt; F-FDG-PET Micro / CT

This paper describes the use of 18F-FDG and micro-PET/CT imaging to determine in vivo glucose metabolism kinetics in mice (and is transferable to rats). ...

Induction and Micro-CT Imaging of Cerebral Cavernous Malformations in Mouse Model

Mutations in the CCM1 (aka KRIT1), CCM2, or CCM3 (aka PDCD10) gene cause cerebral cavernous malformation (CCM) in humans. Mouse models of CCM disease have been established by tamoxifen induced deletion of Ccm genes in postnatal animals. These mouse models provide invaluable tools to investigate molecular mechanism and therapeutic approaches for CCM disease. An accurate and quantitative method to assess lesion burden and progression is essential to harness the full value of these animal models. Here, we demonstrate the induction of CCM disease in a mouse model and the use of the contrast enhanced X-ray micro computed tomography (micro-CT) method to measure CCM lesion burden in mouse brains. At postnatal day 1 (P1), we used 4-hydroxytamoxifen (4HT) to activate Cre recombinase activity from the Cdh5-CreErt2 transgene to cleave the floxed allele of Ccm2. CCM lesions in mouse brains were analyzed at P8. For micro-CT, iodine based Lugol&#39;s solution was used to enhance contrast in brain tissue. We have optimized the scan parameters and utilized a voxel dimension of 9.5 &#181;m, which lead to a minimum feature size of approximately 25 &#181;m. This resolution is sufficient to measure CCM lesion volume and number globally and accurately, and provide high-quality 3-D mapping of CCM lesions in mouse brains. This method enhances the value of the established mouse models to study the molecular basis and potential therapies for CCM and other cerebrovascular diseases.

This protocol demonstrates the induction of cerebral cavernous malformation disease in a mouse model and uses contrast enhanced micro computed tomography to measure lesion burden. This method enhances the value of established mouse models to study the molecular basis and potential therapies for cerebral cavernous malformation and other cerebrovascular diseases.

Induction et Micro-CT Imaging of Cerebral Cavernous Malformations dans le modèle murin

Mutations in the CCM1 (aka KRIT1), CCM2, or CCM3 (aka PDCD10) gene cause cerebral cavernous malformation (CCM) in humans. Mouse models of CCM disease have ...

Quantitative Micro-CT Analysis of Aortopathy in a Mouse Model of &#946;-aminopropionitrile-induced Aortic Aneurysm and Dissection

Aortic aneurysm and dissection is associated with significant morbidity and mortality in the population and can be highly lethal. While animal models of aortic disease exist, in vivo imaging of the vasculature has been limited. In recent years, micro-computerized tomography (micro-CT) has emerged as a preferred modality for imaging both large and small vessels both in vivo and ex vivo. In conjunction with a method of vascular casting, we have successfully used micro-CT to characterize the frequency and distribution of aortic pathology in &#946;-aminopropionitrile-treated C57/Bl6 mice. Technical limitations of this method include variations in the quality of the perfusion introduced by poor animal preparation, the application of proper methodologies for vessel size quantification, and the non-survivability of this procedure. This article details a methodology for the intravascular perfusion of a lead-based radiopaque silicone rubber for the quantitative characterization of aortopathy in a mouse model of aneurysm and dissection. In addition to visualizing aortic pathology, this method may be used for examining other vascular beds in vivo or vascular beds removed post-mortem.

Quantitative Micro-CT Analysis of Aortopathy in a Mouse Model of &amp;#946;-aminopropionitrile-induced Aortic Aneurysm and Dissection

This article describes a detailed methodology of using a radiopaque lead-based silicone rubber to perfuse the murine vasculature for aortic diameter quantification in a mouse model of aortic aneurysm and dissection.

Analyse quantitative de Micro-CT de Aortopathy dans un modèle murin d’anévrisme de l’aorte induite par la β-aminopropionitrile et Dissection

Aortic aneurysm and dissection is associated with significant morbidity and mortality in the population and can be highly lethal. While animal models of ...

Dual Bioluminescence Imaging of Tumor Progression and Angiogenesis

Angiogenesis, as a crucial process of tumor progression, has become a research hotspot and target of anti-tumor therapy. However, there is no reliable model for tracing tumor progression and angiogenesis simultaneously in a visual and sensitive manner. Bioluminescence imaging displays its unique superiority in living imaging due to its advantages of high sensitivity, strong specificity, and accurate measurement. Presented here is a protocol to establish a tumor-bearing mouse model by injecting a Renilla luciferase-labeled murine breast cancer cell line 4T1 into the transgenic mouse with angiogenesis-induced Firefly luciferase expression. This mouse model provides a valuable tool to simultaneously monitor tumor progression and angiogenesis in real-time by dual bioluminescence imaging in a single mouse. This model may be widely applied in anti-tumor drug screening and oncology research.

This protocol describes the establishment of a tumor-bearing mouse model to monitor tumor progression and angiogenesis in real-time by dual bioluminescence imaging.

Double imagerie de bioluminescence de la progression tumorale et de l'angiogenèse

Angiogenesis, as a crucial process of tumor progression, has become a research hotspot and target of anti-tumor therapy. However, there is no reliable ...

Evaluation of Capnography Sampling Line Compatibility and Accuracy when Used with a Portable Capnography Monitor

Capnography is commonly used to monitor patient&#8217;s ventilatory status. While sidestream capnography has been shown to provide a reliable assessment of end-tidal CO2 (ETCO2), its accuracy is commonly validated using commercial kits composed of a capnography monitor and its matching disposable nasal cannula sampling lines. The purpose of this study was to assess the compatibility and accuracy of cross-paired capnography sampling lines with a single portable bedside capnography monitor. A series of 4 bench tests were performed to evaluate the tensile strength, rise time, ETCO2 accuracy as a function of respiratory rate, and ETCO2 accuracy in the presence of supplemental O2. Each bench test was performed using specialized, validated equipment to allow for a full evaluation of sampling line performance. The 4 bench tests successfully differentiated between sampling lines from different commercial sources and suggested that due to increased rise time and decreased ETCO2 accuracy, not all nasal cannula sampling lines provide reliable clinical data when cross-paired with a commercial capnography monitor. Care should be taken to ensure that any cross-pairing of capnography monitors and disposable sampling lines is fully validated for use across respiratory rates and supplemental O2 flow rates commonly encountered in clinical settings.

The goal of this study was to evaluate the accuracy of capnography sampling lines used in conjunction with a portable bedside capnography monitor. Sampling lines from 7 manufacturers were evaluated for tensile strength, rise time, and ETCO2 accuracy as a function of respiratory rate or supplemental oxygen flow rate.

Évaluation de la compatibilité et de l’exactitude de la ligne d’échantillonnage capnographie lorsqu’elle est utilisée avec un moniteur de capnographie portatif

Capnography is commonly used to monitor patient&#8217;s ventilatory status. While sidestream capnography has been shown to provide a reliable assessment ...