Name: in vivo bioluminescent imaging of mammary tumors using ivis spectrum
Uploaded: 2009-04-29T00:00:00.000+00:00
Duration: 8 min 53 s
Description: Watch this Scientific Journal Video about In vivo Bioluminescent Imaging of Mammary Tumors Using IVIS Spectrum at JoVE.com

Uma grande variedade de linhagens celulares expressando luciferase câncer pode ser usado para pesquisa pré-clínica em modelos de mouse. Estas células são fornecidos como uma cultura isentos de agentes patogénicos congelados, que prontamente crescer em mídia padrão sem a necessidade de marcadores de seleção. Para nosso experimento, usaremos o 4T1-luc2 linhagem de células mamárias murinas tumor, que expressa o gene da luciferase, que serve como um indicador óptico de expressão gênica ou tumorgenesis in vivo. Vamos usar a expressão de luciferase para acompanhar o crescimento do tumor primário não-invasiva, mas eles também podem ser usados ​​para localizar e monitorar as lesões metastáticas. Atividade de luciferase deve ser verificada antes da injeção, e para fazer isso, um frasco de 90% confluentes é colhido por tripsinização e então contadas. 50.000 células são, então, dispensadas em um único poço de uma placa de microtitulação e diluições seriadas são realizadas. Luciferina é adicionado aos poços em 150ug por ml e incubadas por 2 minutos. A microplaca pode ser trabalhada em IVIS ou um leitor de placas luminescentes para determinar níveis de expressão. Esta linhagem celular expressa até 6500 fótons por célula por segundo, mas qualquer nível de expressão acima de 500 fótons por célula por segundo pode ser trabalhada com sucesso in vivo. Agora que temos as células de atividade ideal, podemos avançar para a etapa de injeção subcutânea. A fim de facilitar a detecção do tumor ótima, estamos usando uma cepa do mouse atímicos imunocomprometidos nude. Antes da injeção, os animais são anestesiados para efeito de anestesia profunda. Em seguida, vamos injetar até 250.000 células em 100ul PBS são injetadas por via subcutânea no flanco. Carregar as células em uma seringa de 1 ml e anexar uma agulha de calibre 26. Levante a pele suavemente com uma pinça para fazer uma &quot;tenda&quot; e injetar as células na base. As células recém-injetado podem ser visualizados imediatamente. Em cada sessão a imagem de um total de 150mg de luciferina por kg de peso corporal é então administrado através de duas injeções na cavidade peritoneal. Neste estudo, os animais são imaged 10 minutos após a injeção Luciferina para garantir fluxo de fótons consistente. Vamos mostrar isso no próximo passo. Luciferina cinética pode variar de dia para dia. Neste modelo particular, medindo 10 minutos após a injeção Luciferina deu um resultado dentro de uma faixa de variação de 15%. Para nosso experimento, usaremos o Spectrum IVIS in vivo sistema de imagem usa um back-diluído dispositivo de carga acoplado resfriado a -90 ° C para atingir o máximo de sensibilidade. Para apoiar a quantificação absoluta, o sistema de medidas de carga escuro durante o tempo e executa um auto-calibração durante a inicialização. Para iniciar a imagem, inicializar o sistema IVIS (um clique) e definir os parâmetros de imagem para o experimento. Selecione o campo de visão para o número de animais com imagens. Até 5 animais podem ser mantidos no instrumento usando as múltiplas integrante anestésico. O palco está em um constante 37 ° C para manter a temperatura corporal nos animais. Uma porta de ECG é fornecido para monitorar a saúde dos animais durante os procedimentos estressantes. In Living software de imagem, tempo de exposição, f-stop e pixel binning pode ser otimizado com base no nível de expressão da linha celular. Essas configurações podem ser alteradas a qualquer momento durante um experimento sem impactar o resultado quantitativo. IVIS adquire uma imagem fotográfica do animal sob luz branca e um sinal bioluminescente quantitativa ou fluorescentes, que é sobreposta à imagem. O sinal bioluminescente é expressa em fótons por segundo e exibido como um mapa da intensidade. A visualização da imagem é ajustado para oferecer melhor contraste e resolução da imagem sem afetar a quantificação. Luminescência das células pode ser medido no local da injecção com uma região de interesse ferramenta. Dados de medição são exibidos na tabela em conjunto com todos os parâmetros experimentais relacionadas com a captura de imagens que podem ser salvos ou exportados para análise Várias imagens podem ser adquiridos e comparados em estudos longitudinais que cobrem segundos ou meses, dependendo da natureza do experimento. Vamos medir fluxo de fótons a partir do tumor no tempo zero e monitor para 4 semanas com imagens em intervalos quinzenais. Photon fluxo do tumor é proporcional ao número de células vivas para expressar luciferase bioluminescência se correlaciona diretamente com o tamanho do tumor. Em 5 dias após a implantação do tumor ainda não é palpável, mas as células podem ser quantificados através da bioluminescência eo tumor é visto como crescendo ativamente. Nesta fase, o sinal de bioluminescência é muito stronger. O tempo de exposição, f-stop e binning pixel pode ser ajustado para que a imagem é clara ea câmera não saturar. IVIS compensa automaticamente as mudanças na coleta de luz para essas medições podem ser comparados com aqueles coletados antes e depois do experimento. Aos 7 dias pós-implantação tumores são palpáveis, pela primeira vez e bioluminescência medida já gerou sete dias de dados. Aos 28 dias pós-implantação, os tumores estão tornando-se necrótico e células começam a morrer. Tamanho do tumor estimado por pinça de medição não se altera significativamente, mas a luminescência do tumor diminui indicando a morte celular. Medições Caliper e medição da bioluminescência pode ser continuado até uma extremidade humana é atingido. Necrose tumoral devido à hipóxia ou regimes de tratamento será indicado pela bioluminescência reduzido mesmo se eles não reduzem a massa tumoral.

Gostaria de assegurar que todos os experimentos realizados para produzir o artigo do Journal of Experiments visualizada foram realizadas em cumprimento do Protocolo de animais 060 aprovado pelo Institutional Animal Care e Use Committee (IACUC) a Caliper Life Sciences, Alameda CA Jae Kim, Ph . D., Presidente IACUC, Caliper Life Sciences.

in vivo bioluminescent imaging of mammary tumors using ivis spectrum

Camundongo 4T1 células tumorais mamárias pode ser implantado sub-cutânea em nu / nu ratos para formar tumores palpáveis ​​em 15 a 20 dias. Este sistema modelo xenoenxerto tumor é valioso para o pré-clínicos na avaliação in vivo de compostos antitumor putativa. A linhagem celular 4T1 foi projetado para constitutivamente expressam o gene luciferase do firefly (luc2). Quando ratos portadores 4T1-luc2 tumores são injetados com Luciferina os tumores emitir um sinal luminoso visual que pode ser monitorado usando um sistema de imagens ópticas sensíveis como o Spectrum IVIS. O fluxo de fótons a partir do tumor é proporcional ao número de células emissores de luz eo sinal pode ser medido para monitorar o crescimento do tumor e do desenvolvimento. IVIS é calibrado para permitir a quantificação absoluta do sinal bioluminescente e estudos longitudinais podem ser realizadas ao longo de muitos meses e mais de várias ordens de magnitude de sinal, sem comprometer o resultado quantitativo. O crescimento do tumor pode ser monitorado durante vários dias pela bioluminescência antes do tamanho do tumor se torna palpável ou mensurável por meios físicos tradicionais. Esse monitoramento rápido pode fornecer informações sobre eventos precoces no desenvolvimento do tumor ou levar a menor procedimentos experimentais. Morte da célula tumoral e necrose devido ao tratamento hipóxia ou medicamento está indicado no início por uma redução no sinal bioluminescente. Esta morte celular pode não ser acompanhada por uma redução no tamanho do tumor medido por meios físicos. A capacidade de ver eventos no início de necrose tumoral tem um impacto significativo sobre a seleção e desenvolvimento de agentes terapêuticos. Imagens quantitativas do crescimento tumoral utilizando IVIS fornece a quantificação precisa e acelera o processo experimental para gerar resultados.

Watch this Scientific Journal Video about In vivo Bioluminescent Imaging of Mammary Tumors Using IVIS Spectrum at JoVE.com

In vivo Bioluminescent Imaging of Mammary Tumors Using IVIS Spectrum

Células tumorais mamárias expressar luciferase são implantados subcutaneamente em camundongos e visualizado utilizando imagens ópticas para monitorar o crescimento do tumor e de desenvolvimento não-invasiva em um estudo longitudinal.

In vivo Bioluminescent imagem de tumores mamários Usando Spectrum IVIS

4T1 mouse mammary tumor cells can be implanted sub-cutaneously in nu/nu mice to form palpable tumors in 15 to 20 days. This xenograft tumor model system ...

in-vivo-bioluminescent-imaging-of-mammary-tumors-using-ivis-spectrum

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JoVE Journal

Biology

In vivo Bioluminescent Imaging of Mammary Tumors Using IVIS Spectrum

47.0K visualizações.  Caliper Life Sciences.  中文 

Vídeo: In vivo Bioluminescent Imaging of Mammary Tumors Using IVIS Spectrum

Establishing Intracranial Brain Tumor Xenografts With Subsequent Analysis of Tumor Growth and Response to Therapy using Bioluminescence Imaging

Transplantation models using human brain tumor cells have served an essential function in neuro-oncology research for many years. In the past, the most commonly used procedure for human tumor xenograft establishment consisted of the collection of cells from culture flasks, followed by the subcutaneous injection of the collected cells in immunocompromised mice. Whereas this approach still sees frequent use in many laboratories, there has been a significant shift in emphasis over the past decade towards orthotopic xenograft establishment, which, in the instance of brain tumors, requires tumor cell injection into appropriate neuroanatomical structures. Because intracranial xenograft establishment eliminates the ability to monitor tumor growth through direct measurement, such as by use of calipers, the shift in emphasis towards orthotopic brain tumor xenograft models has necessitated the utilization of non-invasive imaging for assessing tumor burden in host animals. Of the currently available imaging methods, bioluminescence monitoring is generally considered to offer the best combination of sensitivity, expediency, and cost. Here, we will demonstrate procedures for orthotopic brain tumor establishment, and for monitoring tumor growth and response to treatment when testing experimental therapies.

Luciferase-modified human brain tumor xenografts can be established intracranially in athymic mice, with subsequent monitoring of tumor growth and response to therapy using bioluminescence imaging. In combination with survival analysis, bioluminescence monitoring is an essential research tool for pre-clinical testing of therapies being considered for treating brain tumors.

Estabelecer xenoenxertos tumor intracraniano Cérebro com posterior análise do crescimento tumoral e resposta à terapia com imagem Bioluminescência

Transplantation models using human brain tumor cells have served an essential function in neuro-oncology research for many years. In the past, the most ...

Neurociência

Monitoring Tumor Metastases and Osteolytic Lesions with Bioluminescence and 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 imaging using IVIS Spectrum was employed to monitor the distribution and development of the tumor cells following the delivery procedure including DLIT reconstruction to measure the tumor signal and its location.
Development of metastatic lesions to the bone tissues triggers osteolytic activity and lesions to tibia and femur were evaluated longitudinally using micro CT. Imaging was performed using a Quantum FX micro CT system with fast imaging and low X-ray dose. The low radiation dose allows multiple imaging sessions to be performed with a cumulative X-ray dosage far below LD50. A mouse imaging shuttle device was used to sequentially image the mice with both IVIS Spectrum and Quantum FX achieving accurate animal positioning in both the bioluminescence and CT images. The optical and CT data sets were co-registered in 3-dimentions using the Living Image 4.1 software. This multi-mode approach allows close monitoring of tumor growth and development simultaneously with osteolytic activity.

An experimental mouse model of bone metastasis was established following intracardiac delivery of luciferase expressing mammary tumor cells. Tumor development and resulted osteolytic lesion were monitored longitudinally with bioluminescence and micro CT imaging.

Monitoramento metástases tumorais e lesões osteolíticas com bioluminescência e CT Micro Imagem

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

Medicina

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.

Na imagem de bioluminescência vivo Tumor de mama Dynamics Hipóxia metástase do cancro cerebral em um modelo de rato

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

In vivo Dual Substrate Bioluminescent Imaging

Our understanding of how and when breast cancer cells transit from established primary tumors to metastatic sites has increased at an exceptional rate since the advent of in vivo bioluminescent imaging technologies 1-3. Indeed, the ability to locate and quantify tumor growth longitudinally in a single cohort of animals to completion of the study as opposed to sacrificing individual groups of animals at specific assay times has revolutionized how researchers investigate breast cancer metastasis. Unfortunately, current methodologies preclude the real-time assessment of critical changes that transpire in cell signaling systems as breast cancer cells (i) evolve within primary tumors, (ii) disseminate throughout the body, and (iii) reinitiate proliferative programs at sites of a metastatic lesion. However, recent advancements in bioluminescent imaging now make it possible to simultaneously quantify specific spatiotemporal changes in gene expression as a function of tumor development and metastatic progression via the use of dual substrate luminescence reactions. To do so, researchers take advantage for two light-producing luciferase enzymes isolated from the firefly (Photinus pyralis) and sea pansy (Renilla reniformis), both of which react to mutually exclusive substrates that previously facilitated their wide-spread use in in vitro cell-based reporter gene assays 4. Here we demonstrate the in vivo utility of these two enzymes such that one luminescence reaction specifically marks the size and location of a developing tumor, while the second luminescent reaction serves as a means to visualize the activation status of specific signaling systems during distinct stages of tumor and metastasis development. Thus, the objectives of this study are two-fold. First, we will describe the steps necessary to construct dual bioluminescent reporter cell lines, as well as those needed to facilitate their use in visualizing the spatiotemporal regulation of gene expression during specific steps of the metastatic cascade. Using the 4T1 model of breast cancer metastasis, we show that the in vivo activity of a synthetic Smad Binding Element (SBE) promoter was decreased dramatically in pulmonary metastasis as compared to that measured in the primary tumor 4-6. Recently, breast cancer metastasis was shown to be regulated by changes within the primary tumor microenvironment and reactive stroma, including those occurring in fibroblasts and infiltrating immune cells 7-9. Thus, our second objective will be to demonstrate the utility of dual bioluminescent techniques in monitoring the growth and localization of two unique cell populations harbored within a single animal during breast cancer growth and metastasis.

In vivo Dual Substrate Bioluminescent Imaging

Herein we describe the methods to construct, visualize, and quantify the bioluminescent reactions of both firefly and renilla luciferase enzymes expressed in metastatic breast cancer cells during their growth and metastasis in vivo.

In vivo dupla Substrato Bioluminescent imagem

Our understanding of how and when breast cancer cells transit from established primary tumors to metastatic sites has increased at an exceptional rate ...

Intracranial Implantation with Subsequent 3D In Vivo Bioluminescent Imaging of Murine Gliomas

The mouse glioma 261 (GL261) is recognized as an in vivo model system that recapitulates many of the features of human glioblastoma multiforme (GBM). The cell line was originally induced by intracranial injection of 3-methyl-cholantrene into a C57BL/6 syngeneic mouse strain 1; therefore, immunologically competent C57BL/6 mice can be used. While we use GL261, the following protocol can be used for the implantation and monitoring of any intracranial mouse tumor model. GL261 cells were engineered to stably express firefly luciferase (GL261-luc). We also created the brighter GL261-luc2 cell line by stable transfection of the luc2 gene expressed from the CMV promoter. C57BL/6-cBrd/cBrd/Cr mice (albino variant of C57BL/6) from the National Cancer Institute, Frederick, MD were used to eliminate the light attenuation caused by black skin and fur. With the use of albino C57BL/6 mice; in vivo imaging using the IVIS Spectrum in vivo imaging system is possible from the day of implantation (Caliper Life Sciences, Hopkinton, MA). The GL261-luc and GL261-luc2 cell lines showed the same in vivo behavior as the parental GL261 cells. Some of the shared histological features present in human GBMs and this mouse model include: tumor necrosis, pseudopalisades, neovascularization, invasion, hypercellularity, and inflammation 1.
Prior to implantation animals were anesthetized by an intraperitoneal injection of ketamine (50 mg/kg), xylazine (5 mg/kg) and buprenorphine (0.05 mg/kg), placed in a stereotactic apparatus and an incision was made with a scalpel over the cranial midline. A burrhole was made 0.1mm posterior to the bregma and 2.3mm to the right of the midline. A needle was inserted to a depth of 3mm and withdrawn 0.4mm to a depth of 2.6mm. Two &mu;l of GL261-luc or GL261-luc2 cells (107 cells/ml) were infused over the course of 3 minutes. The burrhole was closed with bonewax and the incision was sutured.
Following stereotactic implantation the bioluminescent cells are detectable from the day of implantation and the tumor can be analyzed using the 3D image reconstruction feature of the IVIS Spectrum instrument. Animals receive a subcutaneous injection of 150&mu;g luciferin /kg body weight 20 min prior to imaging. Tumor burden is quantified using mean tumor bioluminescence over time. Tumor-bearing mice were observed daily to assess morbidity and were euthanized when one or more of the following symptoms are present: lethargy, failure to ambulate, hunched posture, failure to groom, anorexia resulting in &gt;10% loss of weight. Tumors were evident in all of the animals on necropsy.

Intracranial Implantation with Subsequent 3D In Vivo Bioluminescent Imaging of Murine Gliomas

Intracranial implantation of GL261 cells into C57BL/6 mice produces malignant gliomas that recapitulate many of the hallmarks of human glioblastoma multiforme. We used GL261 cells stably expressing luciferase to allow us to use in vivo imaging to follow tumor progression. The surgery and 3D in vivo imaging are demonstrated.

Implantação intracraniana com 3D subseqüentes In Vivo Bioluminescent imagem de Gliomas murino

The mouse glioma 261 (GL261) is recognized as an in vivo model system that recapitulates many of the features of human glioblastoma multiforme (GBM). The ...

Bioluminescent Bacterial Imaging In Vivo

This video describes the use of whole body bioluminesce imaging (BLI) for the study of bacterial trafficking in live mice, with an emphasis on the use of bacteria in gene and cell therapy for cancer. Bacteria present an attractive class of vector for cancer therapy, possessing a natural ability to grow preferentially within tumors following systemic administration. Bacteria engineered to express the lux gene cassette permit BLI detection of the bacteria and concurrently tumor sites. The location and levels of bacteria within tumors over time can be readily examined, visualized in two or three dimensions. The method is applicable to a wide range of bacterial species and tumor xenograft types. This article describes the protocol for analysis of bioluminescent bacteria within subcutaneous tumor bearing mice. Visualization of commensal bacteria in the Gastrointestinal tract (GIT) by BLI is also described. This powerful, and cheap, real-time imaging strategy represents an ideal method for the study of bacteria in vivo in the context of cancer research, in particular gene therapy, and infectious disease. This video outlines the procedure for studying lux-tagged E. coli in live mice, demonstrating the spatial and temporal readout achievable utilizing BLI with the IVIS system.

Bioluminescent Bacterial Imaging In Vivo

This article describes the administration of lux-tagged bacteria to mice and subsequent in vivo analysis using IVIS bioluminescence imaging.

Imagem bacteriana Bioluminescent<em&gt; Em Vivo</em

This video describes the use of whole body bioluminesce imaging (BLI) for the study of bacterial trafficking in live mice, with an emphasis on the use of ...

Imunologia e Infecção

Monitoring Breast Cancer Growth and Metastatic Colony Formation in Mice using Bioluminescence

Breast cancer is a frequent heterogeneous malignancy and the second leading cause of mortality in women, mainly due to distant organ metastasis. Several animal models have been generated, including the widely used orthotopic mouse models, where cancer cells are injected into the mammary fat pad. However, these models cannot help monitor tumor growth kinetics and metastatic colonization. Cutting-edge tools to monitor cancer cells in real time in mice will significantly advance the understanding of tumor biology. 
Here, breast cancer cell lines stably expressing luciferase and green fluorescent protein (GFP) were established. Specifically, this technique contains two sequential steps initiated by measuring the luciferase activity in vitro and followed by the implantation of the cancer cells into mammary fat pads of nonobese diabetic-severe combined immunodeficiency (NOD-SCID) mice. After the injection, both the tumor growth and metastatic colonization are monitored in real time by the noninvasive bioluminescence imaging system. Then, the quantification of GFP-expressing metastases in the lungs will be examined by fluorescence microscopy to validate the observed bioluminescence results. This sophisticated system combining luciferase and fluorescence-based detection tools evaluates cancer metastasis in vivo, which has great potential for use in breast cancer therapeutics and disease management.

Here, we describe a noninvasive monitoring method involving luciferase and green fluorescent protein expression in various breast cancer cell lines. This protocol provides a technique to monitor tumor formation and metastatic colonization in real time in mice.

Monitoramento do crescimento do câncer de mama e formação de colônia metastática em camundongos usando bioluminescência

Breast cancer is a frequent heterogeneous malignancy and the second leading cause of mortality in women, mainly due to distant organ metastasis. Several ...

Pesquisa do câncer

In Vivo Imaging to Measure Spontaneous Lung Metastasis of Orthotopically-injected Breast Tumor Cells

Metastasis remains the primary cause of cancer-related death. The succession of events that characterize the metastatic cascade presents multiple opportunities for therapeutic intervention, and the ability to accurately model them in mice is critical to evaluate their effects. Here, a step-by-step protocol is presented for the establishment of orthotopic primary breast tumors and the subsequent monitoring of the establishment and growth of metastatic lesions in the lung using in vivo bioluminescence imaging. This methodology allows for the evaluation of treatment or its biological effects along the entire range of metastatic development, from primary tumor escape to outgrowth in the lungs. Breast orthotopic tumors are generated in mice via injection of a luciferase-labeled cell suspension in the 4th mammary gland. Tumors are allowed to grow and disseminate for a specific amount of time and are then surgically resected. Upon resection, spontaneous lung metastasis is detected, and the growth over time is monitored using in vivo bioluminescence imaging. At the desired experimental endpoint, lung tissue can be collected for downstream analysis. The treatment of established, clinically evident metastasis is critical to improve outcomes for stage IV cancer patients, and it can be evaluated through tail vein models of experimental lung metastasis. However, metastatic dissemination occurs early in breast cancer, and many patients have latent, subclinical disseminated disease after surgery. Utilization of spontaneous models such as this one provides the opportunity to study the whole spectrum of the disease, especially the systemic effects driven by treatment of the primary tumor such as pre-metastatic niche priming, and evaluate treatments on dormant and subclinical disease after surgery.

<meta charset="utf8"></head><body>Imagem in vivo para medir metástases pulmonares espontâneas de células tumorais de mama injetadas ortotopicamente

The manuscript describes a methodology for the establishment as well as longitudinal growth monitoring of spontaneous lung metastasis from orthotopically-injected breast tumors, amenable to intervention at all stages of the metastatic cascade.

In vivo Imagem para medir metástase pulmonar espontânea de células tumorais de mama injetadas ortotopicamente

Metastasis remains the primary cause of cancer-related death. The succession of events that characterize the metastatic cascade presents multiple ...

Combined In vivo Optical and &#181;CT Imaging to Monitor Infection, Inflammation, and Bone Anatomy in an Orthopaedic Implant Infection in Mice

Multimodality imaging has emerged as a common technological approach used in both preclinical and clinical research. Advanced techniques that combine in vivo optical and &#956;CT imaging allow the visualization of biological phenomena in an anatomical context. These imaging modalities may be especially useful to study conditions that impact bone. In particular, orthopaedic implant infections are an important problem in clinical orthopaedic surgery. These infections are difficult to treat because bacterial biofilms form on the foreign surgically implanted materials, leading to persistent inflammation, osteomyelitis and eventual osteolysis of the bone surrounding the implant, which ultimately results in implant loosening and failure. Here, a mouse model of an infected orthopaedic prosthetic implant was used that involved the surgical placement of a Kirschner-wire implant into an intramedullary canal in the femur in such a way that the end of the implant extended into the knee joint. In this model, LysEGFP mice, a mouse strain that has EGFP-fluorescent neutrophils, were employed in conjunction with a bioluminescent Staphylococcus aureus strain, which naturally emits light. The bacteria were inoculated into the knee joints of the mice prior to closing the surgical site. In vivo bioluminescent and fluorescent imaging was used to quantify the bacterial burden and neutrophil inflammatory response, respectively. In addition, &#956;CT imaging was performed on the same mice so that the 3D location of the bioluminescent and fluorescent optical signals could be co-registered with the anatomical &#956;CT images. To quantify the changes in the bone over time, the outer bone volume of the distal femurs were measured at specific time points using a semi-automated contour based segmentation process. Taken together, the combination of in vivo bioluminescent/fluorescent imaging with &#956;CT imaging may be especially useful for the noninvasive monitoring of the infection, inflammatory response and anatomical changes in bone over time.

Combined In vivo Optical and  &#181;CT Imaging to Monitor Infection, Inflammation, and Bone Anatomy in an Orthopaedic Implant Infection in Mice

Combined optical and &#956;CT imaging in a mouse model of orthopaedic implant infection, utilizing a bioluminescent engineered strain of Staphylococcus aureus, provided the capability to noninvasively and longitudinally monitor the dynamics of the bacterial infection, as well as the corresponding inflammatory response and anatomical changes in the bone.

Combinado<em&gt; In vivo</em&gt; Optical e μCT imagem para monitorar infecções, inflamações, and Bone Anatomy em uma infecção Orthopaedic Implant em Ratos

Multimodality imaging has emerged as a common technological approach used in both preclinical and clinical research. Advanced techniques that combine in ...

In vivo Bioluminescent imagem de tumores mamários Usando Spectrum IVIS

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