L&#39;imaging longitudinale è usato in studi pre-clinici di seguire i progressi di una malattia o di misurare l&#39;effetto di una terapia. In oncologia, metodi ottici fornire strumenti rigorosi per monitorare la crescita tumorale e fornire quantificazione precisa della crescita cellulare o l&#39;espressione genica in ogni punto in questo studio. Cambiamenti anatomici può essere misurata utilizzando una tecnica ad alta risoluzione come microCT, ma per l&#39;imaging longitudinale a bassa dose di raggi X deve essere utilizzato per evitare artefatti biologici. Immagini ottiche e microCT possono essere co-registrazione per fornire una combinazione di dati funzionali ed anatomiche garantire che il massimo di informazioni viene estratta dal modello animale. 1. Preparazione delle cellule <ol><li> Caliper offre una gamma di linee di cellule tumorali che esprimono luciferasi di ricerca pre-clinica in modelli murini. </li><li> MDA-MB-231-luc-D2H2LN è un essere umano mammario linea di cellule tumorali che esprimono il gene della luciferasi, che può essere utilizzato come indicatore ottico di tumorgenesis in vivo. Questa linea cellulare è stato creato da una metastasi linfonodali spontanea che ha avuto origine da un tumore mammario D3H1 cuscinetto adiposo ed è noto per formare metastasi aggressivo. </li><li> Le cellule sono forniti come un esenti da organismi patogeni cultura congelati che cresce facilmente in supporti standard senza la necessità di marcatori di selezione. </li><li> Per verificare l&#39;attività luciferasi prima iniezione l&#39;animale un pallone al 90% confluenti viene raccolto da tripsinizzazione. Attività luciferasi è misurata dal dosaggio 50.000 cellule in una micropiastra e l&#39;esecuzione di diluizione in serie. </li></ol> 2. Iniezione intracardiaca delle cellule in animali <ol><li> Prima dell&#39;iniezione, gli animali sono anestetizzati con isoflurano 3%. </li><li> 1-3000000 cellule vengono iniettate in volume nel 50ul al ventricolo sinistro. Sospensione cellulare contiene 150ug/ml D-Luciferin per convalidare tecnica di iniezione. </li><li> Per confermare una corretta topi iniezione intracardiaca sono ripreso nel sistema IVIS. I topi che mostrano segnali BLI in tutto il corpo sono state iniettate in modo corretto. Se il segnale è localizzato le cellule non sono stati iniettati correttamente nel cuore. </li></ol> 3. BLI di imaging per il monitoraggio metastasi <ol><li> La navetta di imaging supporta il mouse per l&#39;acquisizione di dati in entrambe le Spectrum IVIS e il sistema QuantumFX microCT. </li><li> La navetta aiuta a mantenere l&#39;animale nella stessa posizione per l&#39;imaging e include i segnali fiduciali per aiutare in automatico co-registrazione. La camera è collegata al vaporizzatore isofluorance per mantenere l&#39;anestesia. </li><li> Imaging guidata nel software immagine vivente 4.1 può essere usato per selezionare automaticamente il tempo di esposizione, f-stop e binning e garantire la raccolta quantitativa dei dati. </li><li> Più immagini possono essere acquisite e confrontati negli studi longitudinali relativi secondi o mesi a seconda della natura di questo esperimento. </li><li> 6 ° giorno dopo l&#39;iniezione di cellule, BLI imaging con IVIS spettro mostra metastasi alle ossa in sette su nove topi. </li></ol> 4. Monitoraggio longitudinale delle lesioni osteolitiche utilizzando micro CT <ol><li> I topi che hanno mostrato le metastasi ossee chiaro visibile utilizzando BLI hanno ripreso nel sistema FX Quantum micro CT. </li><li> Il Quantum FX micro sistema CT offre immagini di alta qualità a raggi X a basso dosaggio tale da consentire vero longitudinale micro CT negli studi preclinici. </li><li> Degradazione delle ossa in entrambe le gambe può essere longitudinalmente monitorati utilizzando questo sistema in meno di un minuto. </li><li> I topi vengono anestetizzati con isoflurano 3% e possono essere trasferiti direttamente dal sistema Spectrum IVIS. </li><li> Due immagini 60 millimetri sono cuciti insieme per catturare tutta la visione degli animali. </li><li> Un FOV 30 mm è ideale per l&#39;imaging del ginocchio in alta risoluzione. </li><li> Una singola scansione ad alta risoluzione sufficiente per misurare e quantificare le metastasi ossee richiede appena 18 secondi e rilascia una dose di 13mGy solo. </li><li> Topi sono stati ripreso fino a 8 volte in 36 giorni con il sistema di micro-CT senza compromettere la crescita del tumore o la salute dell&#39;animale. </li><li> Il micro TC ha mostrato che leisions ossa erano visibili il giorno 10 e progredito molto rapidamente, corrispondente all&#39;aumento del segnale BLI. </li></ol> 5. 3D co-registrazione e analisi dei BLI e UCT <ol><li> Una volta che le immagini sono state catturate sia sul IVIS spettro e Quantum FX micro CT dati di sistema possono essere co-registrati in una ricostruzione 3D. </li><li> MicroCT immagini vengono esportate come file DICOM e caricati nel Living immagine 4,1 strumento modalità Multi 3D. </li><li> In Software immagine vivente eseguire il programma DLIT a tre dimensioni ricostruire il segnale ottico. </li><li> Lo strumento fiduciali nel software viene utilizzato per co-registrare il segnale ottico BLI all&#39;immagine micro CT. </li><li> Coregistered insiemi di dati possono essere analizzati quantitativamente per monitorare i parametri multipli durante la progressione della malattia eccoci monitoraggio della crescita del tumore metastatico insieme con lo sviluppo della lesione osteolitica nell&#39;osso. </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>

Gli autori sono dipendenti di Caliper Life Sciences, che rendono lo strumento descritto in questo articolo.

Di imaging per piccoli animali con microCT è stata applicata al monitoraggio delle lesioni osteolitiche in condizioni patologiche, come lo sviluppo del tumore (Cowey et al 2007;.. Labrinidis et al, 2009). L&#39;imaging bioluminescenza di luciferasi tumori etichettati è stato ampiamente adottato pre-clinico, co-registrazione di immagini bioluminescenza e microCT fornito una migliore definizione della localizzazione anatomica dei segnali bioluminescenza (Kaijzel et al. 2007). Precedente studio con un luciferasi etichettato linea di cellule tumorali della mammella MDA-MB-231-luc-D3H2LN mostrato metastasi alle ossa e lo sviluppo di lesioni osteolitiche dopo l&#39;iniezione intra-cardiaca (Jenkins et al 2005;. Minn et al, 2005). In questo studio, abbiamo analizzato uno studio longitudinale di metastasi tumorali e l&#39;attività osteolitica con combinatoria ottica e basse dosi di radiazioni di imaging microCT con il sistemico MDA-MB-231-luc-D3H2LN modello di metastasi. Abbiamo eseguito l&#39;imaging sequenziale con IVIS Spectrum e Quantum FX sistemi microCT e bioluminescenza raggiunti e CT co-registrazione con la nostra nuova concezione immagine vivente 4,1 software (Heng et al., 2010). Di imaging bioluminescenza con IVIS fornisce rilevamento non invasivo delle cellule tumorali con elevata sensibilità, permettendo di rilevare tumori in stadio precoce. Imaging con Quantum FX permette accurata ricostruzione 3D delle lesioni ossee e dettagliate informazioni anatomiche. Quantum FX sistema offre immagini a bassa dose di radiazioni, che consente il monitoraggio longitudinale di progressione della malattia. Correlazione tra lesioni ossee e delle metastasi del tumore è stato raffigurato in co-registrate immagine, e il vantaggio di monitoraggio multimodalità di sviluppo lesione longitudinale è stato chiaramente dimostrato.

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

A seguito della consegna intracardiaca di MDA-MB-231-luc-D3H2LN cellule Nu / Nu topi, ha sviluppato metastasi sistemiche negli animali iniettati. Di imaging bioluminescenza con Spectrum IVIS è stato impiegato per monitorare la distribuzione e lo sviluppo delle cellule tumorali in seguito alla procedura di consegna tra cui la ricostruzione DLIT per misurare il segnale di tumore e la sua posizione. Sviluppo di lesioni metastatiche ai tessuti ossei innesca l&#39;attività osteolitica e lesioni a tibia e femore sono stati valutati longitudinalmente con micro CT. Imaging è stata eseguita utilizzando un sistema di Quantum FX micro CT con immagini veloci e bassa dose di raggi X. La dose di radiazioni a basso permette sessioni multiple di imaging da eseguire con un cumulativo di raggi X dosaggio molto al di sotto LD50. Un dispositivo di imaging del mouse navetta è stato utilizzato per immagini in sequenza i topi con entrambi Spectrum IVIS e Quantum FX raggiungere un posizionamento preciso animale sia in bioluminescenza e le immagini CT. L&#39;ottica e CT serie di dati sono stati co-registrate in 3-dimentions con il Living immagine 4,1 software. Questo approccio multi-mode permette uno stretto controllo della crescita tumorale e lo sviluppo in contemporanea con l&#39;attività osteolitiche.

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 modello murino sperimentale di metastasi ossee è stata fondata dopo la consegna intracardiaca di cellule tumorali che esprimono luciferasi mammarie. Sviluppo del tumore e ha portato lesione osteolitica sono stati monitorati longitudinalmente con bioluminescenza e micro-TC.

Monitoraggio metastasi tumorali e lesioni osteolitiche con bioluminescenza e 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 ...

monitoring-tumor-metastases-osteolytic-lesions-with-bioluminescence

Research

JoVE Journal

Medicine

24.5K Views.  Caliper Life Sciences.  中文 

Video: 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 Imaging bioluminescenza in vivo di Dinamica Tumore ipossia di metastasi cerebrali cancro al seno in un modello murino

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

Ricerca

Medicina

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.

Osmotica Drug Delivery per arti posteriori ischemico e la perfusione dei vasi con Microfil per Micro-Tomografia Computerizzata Imaging

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.

Bioluminescenza Imaging di un Modello immunocompetenti Animal 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.

Bioluminescenza-Based Metodo Tumore quantificazione per il monitoraggio del tumore Progressione e gli effetti del trattamento in modelli murini linfoma

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.

Imaging fluorescente combinata vicino infrarosso e la tomografia micro-computerizzata per Direttamente Visualizzazione cerebrale tromboembolie

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.

Determinazione metabolismo del glucosio Cinetica Utilizzando<sup&gt; 18</sup&gt; F-FDG micro-PET / TC

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.

Induzione e Micro-CT Imaging delle malformazioni cavernose cerebrali nel modello del topo

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.

Analisi quantitativa di Micro-CT di Aortopathy in un modello murino di indotta da β-aminopropionitrile aneurisma e dissezione

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.

Doppia bioluminescenza Imaging della progressione tumorale e dell'angiogenesi

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.

Valutazione della compatibilità e dell'accuratezza della linea di campionamento capnografica se utilizzata con un monitor capnografico portatile

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