Longitudinal-Bildgebung ist in präklinischen Studien eingesetzt, um den Fortschritt der Krankheit zu folgen oder messen die Wirkung einer therapeutischen. In der Onkologie bieten optische Verfahren rigorose Werkzeuge, um das Tumorwachstum zu überwachen und liefern präzise Quantifizierung des Zellwachstums oder der Genexpression zu jedem Zeitpunkt in einer solchen Studie. Anatomische Veränderungen können mit Hilfe eines hochauflösenden Technik wie MicroCT sein, aber für die Längs-Bildgebung eine niedrige Röntgendosis muss verwendet werden, um biologische Artefakte zu vermeiden. Optische und MicroCT Bilder können co-registriert sein, um eine Kombination von funktionellen und anatomischen Daten sichergestellt wird, dass ein Maximum an Informationen aus dem Tiermodell extrahiert bieten. 1. Zellpräparation <ol><li> Caliper bietet eine Reihe von Luciferase exprimierenden Zelllinien für die präklinische Forschung in Mausmodellen. </li><li> MDA-MB-231-luc-D2H2LN ist eine menschliche Brusttumor-Zelllinie exprimiert das Luciferase-Gen, das als eine optische Anzeige der Tumorgenese in vivo verwendet werden können. Diese Zelllinie ist aus einer spontanen Lymphknotenmetastasen, die von einem D3H1 Brustfettpolster Tumor entstanden ist erstellt und ist dafür bekannt, aggressiv Metastasen bilden. </li><li> Die Zellen werden als pathogen-freien gefrorenen Kultur, die sich leicht wächst in Standard-Medien mit keine Notwendigkeit für Selektionsmarker zur Verfügung gestellt. </li><li> Um zu überprüfen, Luciferaseaktivität vor der Injektion in das Tier eine 90% konfluent Kolben wird durch Trypsinierung geerntet. Luciferaseaktivität wird durch den Verzicht auf 50.000 Zellen in einer Mikrotiterplatte und darstellende Verdünnungsreihe gemessen. </li></ol> 2. Intrakardiale Injektion von Zellen in Tieren <ol><li> Vor der Injektion werden die Tiere narkotisiert mit 3% Isofluran. </li><li> 1 bis 3.000.000 Zellen sind in 50ul Volumen in den linken Ventrikel injiziert. Zell-Suspension enthält 150ug/ml D-Luciferin zu Injektionstechnik zu validieren. </li><li> Zur Bestätigung einer korrekten intrakardiale Injektion Mäuse sind in der IVIS System abgebildet werden. Mäuse, die BLI-Signal zeigen, im ganzen Körper wurden korrekt injiziert. Wenn das Signal lokalisiert wurden die Zellen nicht richtig in das Herz injiziert. </li></ol> 3. BLI Bildgebung zu überwachen Metastasen <ol><li> Die Imaging-Shuttle unterstützt die Maus für die Datenerfassung in beiden IVIS Spectrum und der QuantumFX MicroCT System. </li><li> Der Shuttle hilft bei der Erhaltung des Tieres in der gleichen Position für die Bildgebung und beinhaltet fiducial Signale in automatische Co-Registrierung zu unterstützen. Die Kammer ist der isofluorance Verdampfer verbunden Narkose aufrecht zu erhalten. </li><li> Imaging-Assistenten in der Living Image Software 4.1 kann verwendet werden, um automatisch die Belichtungszeit, Blende und Binning und sorgen für quantitative Datenerhebung werden. </li><li> Mehrere Bilder können erworben werden und im Vergleich in Längsschnittstudien für Sekunden oder Monaten, je nach der Art des Experiments. </li><li> Tag 6 nach der Injektion von Zellen, zeigt BLI Bildgebung mit IVIS Spectrum Metastasen in den Knochen in sieben von neun Mäusen. </li></ol> 4. Longitudinal Monitoring von Osteolysen mit Mikro-CT <ol><li> Mäuse, die klare Knochenmetastasen sichtbar mit BLI zeigten, wurden in den Quantum FX Mikro-CT-System abgebildet. </li><li> Die Quantum FX Mikro-CT-System liefert qualitativ hochwertige Bilder bei einer Röntgen-Dosis niedrig genug, um wahr Längs-Mikro-CT in präklinischen Studien ermöglichen. </li><li> Knochenabbau in beiden Beinen in Längsrichtung überwacht werden mit diesem System in weniger als einer Minute. </li><li> Mäuse sind narkotisiert mit 3% Isofluran und kann direkt aus dem IVIS Spectrum-System übertragen werden. </li><li> Zwei 60mm-Bilder sind zusammen, um das ganze Tier sehen zu erfassen genäht. </li><li> A 30mm FOV ist ideal für die Bildgebung des Kniegelenks in hoher Auflösung. </li><li> Ein einzelner Scan mit ausreichend hoher Auflösung zu messen und zu quantifizieren Knochenmetastasen dauert nur 18 Sekunden und liefert eine Dosis von nur 13mGy. </li><li> Die Mäuse wurden bis zu 8 mal in 36 Tagen mit dem Mikro-CT-System abgebildet, ohne die das Tumorwachstum oder der Gesundheit des Tieres. </li><li> Die Mikro-CT-Bildgebung zeigte, dass Knochen leisions sichtbar am Tag 10 wurden fortgeschritten und sehr schnell, entsprechend der Zunahme der BLI-Signal. </li></ol> 5. 3D-Co-Registrierung und Analyse von BLI und UCT <ol><li> Nachdem die Bilder haben sowohl auf der IVIS Spektrum und Quantum FX Mikro-CT-System Daten erfasst in eine 3D-Rekonstruktion co-registriert werden. </li><li> MicroCT Bilder werden als DICOM-Dateien exportiert und in das Wohnzimmer Bild 4.1 3D Multi Modalität Werkzeug. </li><li> In der Living Image Software laufen DLIT Programm dreidimensional rekonstruieren das optische Signal. </li><li> Die fiducial Tool in der Software wird verwendet, um Co-Registrierung der optischen BLI Signal an die Mikro-CT-Bild. </li><li> Koregistrierten Datensätze können quantitativ analysiert werden, um mehrere Parameter während der Progression der Erkrankung zu überwachen sind wir hier verfolgen das Wachstum der Metastase zusammen mit der Entwicklung der Osteolyse in den Knochen. </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>

Die Autoren sind Mitarbeiter von Caliper Life Sciences, der das Instrument in diesem Artikel vorgestellten machen.

Kleintier-Bildgebung mit MicroCT hat die Überwachung osteolytische Läsionen wurden unter pathologischen Bedingungen, wie Tumor-Entwicklung (..; Labrinidis et al, 2009 Cowey et al 2007) angewendet. Als Biolumineszenz-Bildgebung der Luciferase gekennzeichnet Tumoren weitgehend angewandt worden präklinisch, vorausgesetzt, Co-Registrierung der Biolumineszenz und MicroCT Bilder eine bessere Definition der anatomischen Lage der Biolumineszenz-Signale (Kaijzel et al. 2007). Früheren Studie mit einem Luciferase gekennzeichnet Brusttumor-Zelllinie MDA-MB-231-luc-D3H2LN zeigte Metastasen in Knochen und Entwicklung von Osteolysen folgenden intrakardiale Injektion (Jenkins et al 2005;. Minn et al, 2005). In dieser Studie untersuchten wir eine Längsschnittstudie von Tumormetastasen und osteolytische Aktivität mit Kombinatorik optischen und strahlungsarme-Dosis MicroCT Bildgebung mit der systemischen MDA-MB-231-luc-D3H2LN Metastasen-Modell. Wir führten sequentielle Bildgebung mit IVIS Spectrum und Quantum FX MicroCT Systeme und erreicht Biolumineszenz-und CT-Co-Registrierung mit unserem neu entwickelten Wohnzimmer Bild 4.1-Software (Heng et al., 2010). Biolumineszenz-Bildgebung mit IVIS bietet nicht-invasive Detektion der Tumorzellen mit hoher Empfindlichkeit, so dass die Tumoren in einem frühen Stadium erkannt werden. Imaging mit Quantum FX ermöglicht die genaue 3D-Rekonstruktion der Knochenläsionen und detaillierten anatomischen Informationen. Quantum FX-System bietet niedrige Strahlendosis Bildgebung, die Längs-Überwachung des Krankheitsverlaufs ermöglicht. Korrelation von Knochenläsionen und Tumormetastasen war in der Zusammenarbeit registrierte Bild dargestellt, und der Vorteil der Multimodalität Verfolgung von Läsion Entwicklung längs eindeutig nachgewiesen werden konnte.

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

Nach intrakardialen Lieferung von MDA-MB-231-luc-D3H2LN Zellen Nu / Nu-Mäusen, systemische Metastasen in den injizierten Tieren entwickelt. Biolumineszenz-Bildgebung mit IVIS Spectrum wurde eingesetzt, um den Vertrieb und die Entwicklung der Tumorzellen nach der Lieferung Verfahrens einschließlich DLIT Rekonstruktion des Tumors Signal und seine Lage Maßnahme zu überwachen. Entwicklung von Metastasen an den Knochen Gewebe löst osteolytische Aktivität und Läsionen an Tibia und Femur wurden längs mit Mikro-CT untersucht. Imaging wurde mit einem Quantum FX Mikro-CT-System mit schnellen Bildgebung und niedrige Röntgendosis. Die geringe Strahlendosis ermöglicht mehreren Imaging-Sitzungen mit einer kumulativen X-ray Dosierung weit unter LD50 durchgeführt werden. Eine Maus Imaging-Shuttle-Gerät wurde verwendet, um nacheinander Bild der Mäuse mit beiden IVIS Spectrum und Quantum FX Voraussetzung für präzise Positionierung Tier sowohl in der Biolumineszenz-und CT-Bilder. Die optischen und CT-Datensätze wurden in 3-dimentions co-registriert die Living Bild 4.1-Software. Diese Multi-Mode-Ansatz ermöglicht eine engmaschige Überwachung des Tumorwachstums und der Entwicklung gleichzeitig mit osteolytischen Aktivität.

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

Ein experimentelles Mausmodell von Knochenmetastasen wurde nach intrakardialen Lieferung von Luciferase exprimierenden Mammatumorzellen gegründet. Tumorentstehung und führte Osteolyse wurden in Längsrichtung mit Biolumineszenz-und Mikro-CT-Bildgebung überwacht.

Überwachung Tumormetastasen und Osteolysen mit Biolumineszenz und 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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Serie: 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.

In-vivo-Biolumineszenz-Bildgebung von Tumorhypoxie Dynamics of Breast Cancer Hirnmetastasen in einem Mausmodell

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

Forschung

Medizin

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.

Osmotic Drug Delivery zu ischämischen Hinterbeine und Perfusion von Blutgefäßen mit Microfil für Mikro-Computertomographie

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.

Biolumineszenz-Bildgebung von einem immunkompetenten Tiermodell für Glioblastoma

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.

Biolumineszenz-basierte Tumor Quantifizierung Verfahren zur Überwachung Tumorprogression und Therapieeffekte bei den Maus-Lymphom-Modelle

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.

Kombinierte Near-Infrarot-Fluoreszenz-Bildgebung und Mikrocomputertomographie für Direkt Visualizing Cerebral Thromboembolie

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.

Die Bestimmung Glucose Metabolism Kinetics Verwendung<sup&gt; 18</sup&gt; F-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). ...

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.

Induktion und Mikro-CT-Bildgebung des zerebralen höhlenartigen Fehlbildungen im Mausmodell

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.

Quantitative Micro-CT-Analyse der Aortopathy in einem Mausmodell der β-Aminopropionitrile-induzierte Aortenaneurysma und Dissektion

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.

Dual Biolumineszenz-Bildgebung der Tumorprogression und Angiogenese

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.

Bewertung der Kompatibilität und Genauigkeit der Capnography Sampling Line bei Verwendung mit einem tragbaren Capnographie-Monitor

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