ルシフェラーゼを発現する癌細胞株の多種多様なマウスモデルにおける前臨床研究に使用することができます。 これらの細胞は容易に選択マーカーを必要とせず、標準のメディアが大きくなると思われる無菌凍結培養物として提供されています。 私たちの実験では、我々はin vivoでの遺伝子発現やtumorgenesisの光学指標となるルシフェラーゼ遺伝子を発現4T1 - luc2マウス乳腺腫瘍細胞株を、使用します。我々は、非侵襲的に原発腫瘍の成長を追跡するためにルシフェラーゼ発現を使用しますが、彼らはまた、転移性病変を検索し、監視するために使用することができます。 ルシフェラーゼ活性は、注射する前に検証する必要があります、そしてこれを実行するためには、90％コンフルエントフラスコをトリプシン処理によって回収してからカウントされます。 50,000個の細胞は、マイクロタイタープレートの1ウェルに分注しており、連続希釈が行われる。ルシフェリン、1m​​l当たり150ugでウェルに添加し、2分間インキュベートする。マイクロプレートは、IVISまたは発現レベルを決定するために発光プレートリーダーで画像化することができます。この細胞株は、毎秒のセルあたり6500光子まで表現し、1秒あたりのセル当たり500光子上記のいずれかの発現レベルは、in vivoで正常に画像化することができます。 今我々は最適な活性の細胞を持っていることを、我々は、皮下注射の手順に進むことができます。 腫瘍の最適な検出を容易にするために、我々は胸腺免疫不全ヌードマウス系統を使用しています。注射の前に、動物は深い麻酔のための効果に麻酔する。 次に我々は100ulのPBSに250,000細胞まで注入されることは脇腹に皮下注射されています。 1mlの注射器で細胞をロードし、26ゲージの針を取り付けます。鉗子は&quot;テント&quot;を作り、基地の細胞を注入することでお肌を持ち上げます。新たに注入された細胞は、すぐにイメージングすることができます。各イメージングセッションでは体重kg当たりルシフェリンの150mgの合計は、腹腔内に2回の注射により投与される。 本研究では、動物が一貫した光子束を確保するためにルシフェリンの投与後10分をイメージされています。我々は、次のステップであなたにこれを紹介します。ルシフェリン反応速度は日によって異なることができる。この特定のモデルでは、ルシフェリンの投与後10分を測定することで15％のばらつきの範囲内の結果を与えた。 私たちの実験のために我々は、in vivoイメージングシステムでIVISスペクトラムを使用します裏面入射電荷結合素子を使用して° C最高感度を達成するために-90に冷却。絶対定量をサポートするために、システムはダウンタイム中に暗い電荷を測定し、初期化時にセルフキャリブレーションを実行します。イメージングを開始するには、IVISシステムを初期化する（ワンクリック）と実験用イメージングパラメータを設定します。 撮像される動物の数のビューのフィールドを選択します。最大5匹の動物に不可欠な麻酔のマニホールドを使用して楽器に維持することができる。ステージでは、° C、動物の体温を維持するために一定の37℃です。 EKGポートは、ストレスの多い処置中に動物の健康を監視するために提供されています。 画像ソフトをリビングでは、露光時間、F - stopとピクセルビニングは、細胞株の発現レベルに基づいて最適化することができます。これらの設定は、定量的な結果に影響を与えることなく、実験中にいつでも変更することができます。 IVISは、白色光と画像の上に重ねている定量的な生物発光または蛍光シグナル、下の動物の写真画像を取得。 生物発光シグナルは、秒あたりの光子で表され、強度マップとして表示されます。画像表示は、定量に影響を与えることなく、画像の最適なコントラストと分解能を提供するように調整されます。 細胞からの発光は、関心のツールの域を使用して、注射の部位で測定することができます。測定データは、分析のために保存またはエクスポートすることができます画像のキャプチャに関連するすべての実験パラメータと一緒にテーブルに表示されます複数の画像を取得し、実験の性質に応じて数秒または数か月をカバーする長期的な研究に比較することができます。我々は、隔週の間隔で撮影して4週間の時間がゼロとモニターで腫瘍から光子束を測定します。 腫瘍からのフォトンフラックスは、生物発光は、腫瘍の大きさと直接相関するので、ルシフェラーゼを発現する生細胞の数に比例します。 5日後の移植で腫瘍はまだ明白ではないが、細胞は生物発光によって定量化することができると腫瘍が積極的に成長しているように見られている。この段階での生物発光シグナルは非常にstrongeですR.露光時間は、F - stopとピクセルビニングは、画像が鮮明になるとカメラが飽和しないように調整することができます。これらの測定は実験の前半​​で、後に収集したものに比較することができるように、IVISは、自動的に集光の変化を補正。 7日間でポスト移植腫瘍は、測定値が既にデータの7日間を生成したのは今回が初めてと生物発光のための触知です。 28日のポストの移植で、腫瘍は壊死性になりつつあると細胞が死ぬことを始める。キャリパー測定により推定された腫瘍のサイズがかなり変更されませんが、腫瘍からの発光は、細胞死を示す減少します。 人道的エンドポイントに到達するまで、キャリパーの測定と生物発光の測定を継続することができます。低酸素症または治療体制に起因する腫瘍壊死は、それらが腫瘍塊を削減していない場合でも減少生物発光によって示されます。

私は可視化実験の記事のジャーナルを生成するために実行されるすべての実験は動物実験によって承認された動物のプロトコル060に準拠して行われたことを保証し、キャリパーライフサイエンスにおける委員会（IACUC）、アラメダカリフォルニア州在金は、Phを使用したいと思います。D.、IACUCチェア、キャリパーライフサイエンス。

in vivo bioluminescent imaging of mammary tumors using ivis spectrum

4T1マウス乳癌細胞は、15〜20日に触知腫瘍を形成するために、nu / nuマウスに皮下移植することができる。この異種移植腫瘍モデル系では、推定上の抗腫瘍化合物の in vivo評価における前臨床試験のために貴重です。 4T1細胞株は、構成的にホタルルシフェラーゼ遺伝子（luc2）を発現するように設計されています。 4T1 - luc2腫瘍を持つマウスは、腫瘍がIVISスペクトラムのように敏感な光学イメージングシステムを使用して監視することができる視覚的な光信号を発するルシフェリンを注入されている場合。腫瘍からの光子束は、発光細胞の数に比例し、信号が腫瘍の成長と発展を監視するために測定することができます。 IVISは、何ヶ月にわたって実施し、信号の数桁以上の定量的な結果を損なうことなく可能な生物発光信号と縦断的研究の絶対定量を可能にするために較正されています。 腫瘍の大きさは従来の物理的手段によって触知または測定可能になる前に腫瘍の成長は、生物発光で数日間監視することができます。この急速なモニタリングは、腫瘍の開発の初期段階でのイベントへの洞察を提供したり、短い実験手順につながることができます。 低酸素症または薬物治療による腫瘍細胞死と壊死を​​早期に生物発光信号の減少によって示されます。この細胞死は、物理的手段によって測定された腫瘍の大きさの減少を伴うされていない可能性があります。腫瘍壊死の初期のイベントを表示する機能は、治療薬の選択と開発に大きな影響を与えます。 IVISを用いて腫瘍の増殖の定量的イメージングは​​、正確な定量を提供し、結果を生成する実験的なプロセスを加速させます。

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

ルシフェラーゼを発現する乳腺腫瘍細胞をマウスに皮下移植し、縦断的研究で、非侵襲的に腫瘍の成長と発展を監視する光イメージングを用いて可視化されています。

IVISスペクトラムを用いた乳腺腫瘍の in vivo生物発光イメージングに

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

Universidad San Sebastian

Research

JoVE Journal

Biology

In vivo Bioluminescent Imaging of Mammary Tumors Using IVIS Spectrum

47.0K ビュー.  Caliper Life Sciences.  中文 

ビデオ: 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.

生物発光イメージングを使用して腫瘍の成長や治療に対する反応のその後の分析では頭蓋内脳の腫瘍異種移植片の確立

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

神経科学

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.

生物発光とマイクロCTイメージングで腫瘍転移と溶骨性病変の監視

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

医学

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.

マウスモデルにおける乳がんの脳転移の腫瘍の低酸素動態の in vivo生物発光イメージングに

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.

生体デュアル基質生物発光イメージングにおける

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.

その後の3Dと頭蓋内注入インビボでの</emマウス神経膠腫の>生物発光イメージング

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.

生物発光細菌のイメージング<emインビボで&gt;</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 ...

免疫・感染学

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.

生物発光を用いたマウスにおける乳がん増殖および転移性コロニー形成のモニタリング

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

がん研究

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>同所性注射された乳房腫瘍細胞の自発的な肺転移を測定するためのin vivoイメージング

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.

インビボ 同所注射された乳房腫瘍細胞の自然肺転移を測定するためのイメージング

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.

合わせた<em&gt;インビボ</em&gt;光とμCTイメージングマウスにおける整形外科インプラント感染で感染症、炎症、および骨解剖学を監視する

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

IVISスペクトラムを用いた乳腺腫瘍の in vivo生物発光イメージングに

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生物発光イメージングを使用して腫瘍の成長や治療に対する反応のその後の分析では頭蓋内脳の腫瘍異種移植片の確立

生物発光とマイクロCTイメージングで腫瘍転移と溶骨性病変の監視

マウスモデルにおける乳がんの脳転移の腫瘍の低酸素動態の in vivo生物発光イメージングに

生体デュアル基質生物発光イメージングにおける

その後の3Dと頭蓋内注入インビボでの生物発光イメージング

生物発光細菌のイメージング

生物発光を用いたマウスにおける乳がん増殖および転移性コロニー形成のモニタリング

インビボ同所注射された乳房腫瘍細胞の自然肺転移を測定するためのイメージング

インビボ同所注射された乳房腫瘍細胞の自然肺転移を測定するためのイメージング

合わせた

IVISスペクトラムを用いた乳腺腫瘍の in vivo生物発光イメージングに

文字起こし

要約

さらに動画を探す

この動画の章

生物発光イメージングを使用して腫瘍の成長や治療に対する反応のその後の分析では頭蓋内脳の腫瘍異種移植片の確立

生物発光とマイクロCTイメージングで腫瘍転移と溶骨性病変の監視

マウスモデルにおける乳がんの脳転移の腫瘍の低酸素動態の in vivo生物発光イメージングに

生体デュアル基質生物発光イメージングにおける

その後の3Dと頭蓋内注入インビボでの生物発光イメージング

生物発光細菌のイメージング

生物発光を用いたマウスにおける乳がん増殖および転移性コロニー形成のモニタリング

インビボ 同所注射された乳房腫瘍細胞の自然肺転移を測定するためのイメージング

インビボ 同所注射された乳房腫瘍細胞の自然肺転移を測定するためのイメージング

合わせた

インビボ同所注射された乳房腫瘍細胞の自然肺転移を測定するためのイメージング

インビボ同所注射された乳房腫瘍細胞の自然肺転移を測定するためのイメージング