我々は、米国国立がん研究所（NCI）R01 CA128908とスタンフォード医科学者の研究奨学金からサポートを承諾。この記事に関連する関心の他の潜在的な衝突は報告されなかった。

1。腫瘍モデル<ol><li> 10％ウシ胎児血清および1％ペニシリン/ストレプトマイシン（インビトロジェンライフテクノロジーズ）を補充したRPMI 1640培地で培養H460細胞（アメリカン·タイプ·カルチャー·コレクション）。それは、細胞株、培養培地、接種の場所、マウスあたり異種移植片の数、およびその他の考慮事項の選択肢が特定の研究の目標に合わせて調整するために、すべてのことに留意すべきである。ここでは、イラストだけとして機能するように、ある特定のプロジェクトのデザインを紹介します。 </li><li> 37℃、新鮮培地への変更隔日で5％CO 2の加湿雰囲気下での細胞系を維持している。 </li><li> 75％の細胞がコンフルエントな単層を形成する際には、トリプシンを用いて単層を切り離し、さらに細胞培養のための単一細胞懸濁液に細胞を解離する。 </li><li>年には約1×10 6 H460細胞をサスペンドリン酸緩衝食塩水（PBS; Invitrogen）および皮下にインプラントヌードマウスの左右両肩（雌無胸腺ヌードマウス（ のnu / nu）、4から6週齢、チャールズ·リバー·ラボラトリーズ社）。 </li><li>腫瘍が150まで成長することができます- 200ミリメートル3。それは、このサイズに成長H460腫瘍異種移植片のために約2週間かかります。標準ノギス測定が腫瘍サイズを追跡するために行われる。 </li><li>腫瘍が理想のサイズに達すると担癌マウスは今治療のための準備とPETとCLIの両方を介してin vivoイメージングです。 </li></ol> 2。 PET <ol><li>このスケジュールまたは特定のプロジェクト（ 図1）12に応じてそのいずれかの変動に応じてPET検査を実行します。多くの要因は、スケジュールの設計に影響を含むが、これらに限定されるものではなく、腫瘍異種移植片細胞株、抗癌剤、および投与レジメンの選択でした。ここでは、1つの特定のイメージングのスケジュールを紹介します。 CLIの研究は、に準じて行うことになっているCLIを使用したPET研究と同様のスケジュールでは、対応するPETの直後に行う。また、PET研究の目的は、CLIの結果の検証のため主にであることに留意すべきである。ちょうどイメージング放射性標識プローブのためOIの楽器を使用したい共通のユーザーの場合は、PETは必要ありません。一つは欲望のPET検証を行うとすれば、それは、PETとCLIの楽器が原因で18 Fの半減期が短い（109.77分）の成功するために検証のために非常に近接して内に位置しなければならないことを強調しなければならない。 </li><li>治療群と対照群（n≥3ずつ）にマウスを割ります。 0日目および2で20 mg / kgのベバシズマブの2回注射による治療群のマウスを治療する。 0日目は、最初の注射によって定義されます。デイ-1でプリスキャンは、PETとCLIの両方を介して行われるべきであることに注意してください。 </li><li>担癌マウスの小動物PETはR4のげっ歯類モデルスキャナ（シーメンスメディカルソリューションズ米国で実行される社）。 </li><li> （Aerrane;バクスター）2％イソフルランですべてのマウスを麻酔し、F-fluorothymidine 3&#39;-デオキシ-3&#39;-18を注入する（18、F-FLT、7.3から8.0 MBqの[198から215μCiの]）尾静脈を介して。 PETプローブは、注射前にPBSで希釈される。 </li><li> 1時間後、再びマウスを麻酔し、麻酔したマウスでは発生しやすく、小動物PET装置の視野の中央付近に配置します。 </li><li> 3分間の静的スキャンを取得し、2次元秩序-サブセット期待最大アルゴリズムで画像を再構成する。バックグラウンド補正は必要ありません。 </li><li>減衰補正した全身コロナル画像上の腫瘍の上に、関心領域（冠状および横断スライス用に5ピクセルのROI）を描画します。 ROIのから毎分あたりの最大ピクセル数を取得し、キャリブレーション定数を使用することにより、毎分ミリリットル当たりのカウントに変換されます。 1グラム/ mlの組織の密度を想定して、カウントにROIを変換毎分当たりのグラム。注入量によって毎分グラム当たりのカウント数を割ることによって画像のROI由来％のID / gの値を決定します。減衰補正は必要ありません。 </li></ol> 3。 CLI <ol><li> CLIは、IVISスペクトラムシステム（キャリパーライフサイエンス）を用いて実施されるべきである。画像の取得と分析は、リビングイメージ3.0ソフトウェア（キャリパーライフサイエンス）を用いて行わなければならない。波長分解分光イメージングは​​、18組の狭帯域発光フィルタ（ - 850 nmの490）を使用して実行されるべきである。繰り返しになりますが、各マウスについて、PET研究がプロトコルに含まれている場合、放射性崩壊の量を最小にするためにすぐにPETの後にCLIを実行します。 </li><li>イソフルラン麻酔下で遮光チャンバー内に動物を置きます。複数のマウスは、スループットを向上させるために同時に配置することができます。 </li><li> 3分間の露光時間（F /ストップ= 1、ビニング= 4）を使用して画像を取得する。同じ照明設定（ランプ電圧、フィルター、F /ストップ、視野、BINNを使用る）は、すべての画像を取得する。背景組織の信号強度を計算するために背側の皮膚領域を使用します。ステラジアン当たり平方センチメートル毎秒（P / S / cmの2 / SR）当たりの光子の蛍光発光を正規化します。 </li></ol> 4。代表的な結果 CLIおよびPET画像間の視覚的な比較を容易に行うことができる。同じモダリティと場所側ずつCLIおよびPET画像側から画像全体で統一スケールバーは、この代表的なパネル（ 図2A）、CLIおよびPETの両方が治療前から治療したマウスで、H460異種移植片から大幅に減少した信号を明らかにすることで見ることができた後3日目に、有意な治療効果を示唆している。比較として、適度に変わらない信号に増加同じ期間（データは示さず）中に未処理マウスで観察された。単独で目視により1は、視覚的な腫瘍のコントラストの間に良好な一貫性があることを観察することができますCLIおよびPETから局所化。実際には、この視覚的相関は、抗がん治療レジメン（3日目からのCLIおよびPET画像を比較してください）​​に二次腫瘍の中心壊死を示すのに十分な解像度を持っています。画像所見を検証するために数量化と相関分析を行うことができる。 CLIおよびPET画像の定量化と線形回帰で簡単フィッティングは2種類の治療法が実際に高い相関が得られ（ 図2B、R 2は、18 F-FLTのための= 0.9309、治療群をプローブ）を有していたことが明らかになった。特に、異なる腫瘍モデルと異なる抗がん剤とのCLIおよびPETイメージング研究のすべてに収まるの斜面も著しく接近している、したがって、すべてのデータの線形回帰の優れたフィット感を示唆している（データは示さず）凝集している。双方の代表イメージが我々の前の出版物12を適合させたものです。 er.withinページ= &quot;always&quot;を&gt; <img src="/files/ftp_upload/4341/4341fig1.jpg" alt="図1"/> 図1 PETおよびCLIの研究の実験的な設計の回路図腫瘍は肩領域に両側性に移植され、150〜200ミリメートル3まで成長させ、及び担癌マウスは一日-1、1、3でPETとCLIを介してin vivoイメージングを行ったされた。ベバシズマブ治療は、0日目と2で20 mg / kgを2回注射することにより行った。 <img src="/files/ftp_upload/4341/4341fig2.jpg" alt="図2"/> 図2。 （）in vivoでのCLIおよびベバシズマブで治療したH460異種移植片を有するマウスのPET画像における処理（スキャン前）の前と治療（3日目）の後。CLIおよびPETの結果（n = 3）での（B）に対応する定量分析とその相関。画像は（6）から適応。arge.jpg &quot;ターゲット=&quot; _blank &quot;&gt;拡大図を表示するには、ここをクリックしてください。

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特別な利害関係は宣言されません。

CLIには、多くの基礎科学研究への応用、さらには臨床使用4,5,15,16,17のポテンシャルを発見した有望な分子イメージング技術として浮上している。かなり安価、そしてより広く研究者が利用できる短い取得時間と高いスループットが特徴使いやすくなっていますそのようなOI楽器の使用からPET幹などの伝統的な核イメージングモダリティを介したCLIの主な利点は、。また、どのようなCL...

<table border="1"&gt;<tr&gt;<td<strong&gt;名前</strong</td&gt;<td<strong&gt;会社概要</strong</td&gt;<td<strong&gt;カタログ番号</strong</td&gt;</tr&gt;<tr&gt;<td&gt; H460細胞株</td&gt;<td&gt;アメリカン·タイプ·カルチャー·コレクション</td&gt;<td&gt; ATCC番号：HTB-177</td&gt;</tr&gt;<tr&gt;<td&gt; RPMI 1640培地</td&gt;<td&gt;インビトロジェンライフテクノロジーズ</td&gt;<td&gt; 12633-012</td&gt;</tr&gt;<tr&gt;<td&gt;ウシ胎児血清</td&gt;<td&gt;インビトロジェンライフテクノロジーズ</td&gt;<td&gt; 10091-148</td&gt;</tr&gt;<tr&gt;<td&gt;ペニシリン/ストレプトマイシン</td&gt;<td&gt;インビトロジェンライフテクノロジーズ</td&gt;<td&gt; 15640-055</td&gt;</tr&gt;<tr&gt;<td&gt;リン酸緩衝生理食塩水</td&gt;<td&gt;インビトロジェンライフテクノロジーズ</td&gt;<td&gt; 10010-023</td&gt;</tr&gt;<tr&gt;<td&gt;女性無胸腺ヌードマウス</td&gt;<td&gt;チャールス·リバー株式会社</td&gt;<td&gt;ひずみコード：088</td&gt;</tr&gt;<tr&gt;<td&gt;ベバシズマブ（アバスチン）</td&gt;<td&gt; Genentech / Roche社</td&gt;<td&gt; N / A</td&gt;</tr&gt;<tr&gt;<td&gt; microPETをげっ歯類、R4</td&gt;<td&gt;シーメンスメディカルソリューションズUSA社</td&gt;<td&gt; N / A</td&gt;</tr&gt;<tr&gt;<td&gt;イソフルラン（Aerrane）</td&gt;<td&gt;バクスター</td&gt;<td&gt;バクスター番号：AHN3637</td&gt;</tr&gt;<tr&gt;<td&gt; IVISスペクトラム</td&gt;<td&gt;キャリパーライフサイエンス</td&gt;<td&gt; N / A</td&gt;</tr&gt;</table

cerenkov luminescence imaging (cli) for cancer therapy monitoring

分子イメージングは、陽電子放射断層撮影法（PET）や光イメージング（OI）は1月3日の最も重要なので、最も広く使用されている様式のうちの2つです。 OIは非放射、相対的に低いコスト、短いスキャン時間、高スループット、および基本的な研究者に広く利用のために注目される一方PETは、その優れた感度と定量能力によって特徴づけられる。ただし、両方のモダリティは、同様に自分の欠点を持っています。 OIは主に、その限られた組織の浸透のために生きている組織の厚さを通して著名な散乱光信号と一緒に前臨床アプリケーションに限定されているPETは、貧しい人々の空間分解能と高コストに苦しんでいる。 最近のPETとOIの間に橋がチェレンコフ発光イメージング（CLI）4-6の発見に浮上している。 CLIは、OIの楽器と画像放射性核種にチェレンコフ放射（CR）を活かした新たな画像診断法である。ロシアのノーベルラウルeate Alekseyevichチェレンコフと彼の同僚は、もともと1934年にCRを発見しました。これは、荷電粒子が誘電体媒質7,8で超光速スピードで走行 ​​する際に放出される電磁放射の形です。荷電粒子は、陽電子や電子かどうか、その原子に電子を変位させることにより、媒体の電磁界を乱す。避難電子が基底状態に戻るように崩壊光子の通過した後に放出される。たとえば、1つの18 Fの減衰が水5で3光子の平均を生成すると推定された。 その出現以来、CLIは他人4,5,9,10,11間の生体内腫瘍イメージング、レポーター遺伝子イメージング、放射性トレーサーの開発、マルチモダリティイメージングにおいて 、前臨床含む様々なアプリケーションでの使用のために研究されてきた。 CLIは、これまで多くの成功を収めている理由で最も重要な理由は、この新技術は、低COを利用していることですstとだけPETなどの、より高価であまり利用でき核イメージングモダリティにより結像されていた画像の放射性核種にOIの広い空。 ここでは、がん薬物療法を監視するには、CLIを使用する方法を紹介します。当社グループは、最近、この新しいアプリケーションを調査し、概念実証研究12でその実現可能性を検証しています。我々は、CLIおよびPETは異なる腫瘍異種移植片およびイメージングプローブを越え優れた相関を示したことを明らかにした。これは、CLIは、本質的に、PETと同じ放射性核種を可視化することをCRの包括的な原則と整合的である。それはよく知られている血管新生阻害剤13,14であるため、私たちの治療薬として、我々は、（Genentech / Roche社アバスチン）ベバシズマブ選択。近い将来、この技術の成熟は有意な臨床前医薬品開発への影響、スクリーニング、並びに治療を受けている患者の治療モニタリングを持つように想定することができる。

Watch this Scientific Journal Video about Cerenkov Luminescence Imaging CLI for Cancer Therapy Monitoring at JoVE.com

Cerenkov Luminescence Imaging CLI for Cancer Therapy Monitoring

前臨床癌治療を監視するためのチェレンコフ発光イメージング（CLI）の使用は、ここで説明されます。この方法は、チェレンコフ放射（CR）および放射性標識プローブを可視化する光学イメージング（OI）を利用していますので、前臨床治療モニタリングおよび薬物スクリーニングにおけるPETの代替手段を提供します。

がん治療のモニタリングのためのチェレンコフ発光イメージング（CLI）

In  molecular imaging, positron emission tomography (PET) and optical imaging (OI)  are two of the most important and thus most widely used modalities1-3. ...

cerenkov-luminescence-imaging-cli-for-cancer-therapy-monitoring

Research

JoVE Journal

Medicine

Cerenkov Luminescence Imaging (CLI) for Cancer Therapy Monitoring

13.6K ビュー.  Stanford University. 前臨床癌治療を監視するためのチェレンコフ発光イメージング（CLI）の使用は、ここで説明されます。この方法は、チェレンコフ放射（CR）および放射性標識プローブを可視化する光学イメージング（OI）を利用していますので、前臨床治療モニタリングおよび薬物スクリーニングにおけるPETの代替手段を提供します。

ビデオ: Cerenkov Luminescence Imaging CLI for Cancer Therapy Monitoring

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

Modeling and Imaging 3-Dimensional Collective Cell Invasion

A defining characteristic of cancer malignancy is invasion and metastasis 1. In some cancers (e.g. glioma 2), local invasion into surrounding healthy tissue is the root cause of disease and death. For other cancers (e.g. breast, lung, etc.), it is the process of metastasis, in which tumor cells move from a primary tumor mass, colonize distal sites and ultimately contribute to organ failure, that eventually leads to morbidity and mortality 3. It has been estimated that invasion and metastasis are responsible for 90&#37; of cancer deaths 4. As a result, there has been intense interest in identifying the molecular processes and critical protein mediators of invasion and metastasis for the purposes of improving diagnosis and treatment 5.
A challenge for cancer scientists is to develop invasion assays that sufficiently resemble the in vivo situation to enable accurate disease modeling 6. Two-dimensional cell motility assays are only informative about one aspect of invasion and do not take into account extracellular matrix (ECM) protein remodeling which is also a critical element. Recently, research has refined our understanding of tumor cell invasion and revealed that individual cells may move by elongated or rounded modes 7. In addition, there has been greater appreciation of the contribution of collective invasion, in which cells invade in strands, sheets and clusters, particularly in highly differentiated tumors that maintain epithelial characteristics, to the spread of cancer 8.
We present a refined method 9 for examining the contributions of candidate proteins to collective invasion 10. In particular, by engineering separate pools of cells to express different fluorescent proteins, it is possible to molecularly dissect the activities and proteins required in leading cells versus those required in following cells. The use of RNAi provides the molecular tool to experimentally disassemble the processes involved in individual cell invasion as well as in different positions of collective invasion. In this procedure, mixtures of fluorescently-labeled cells are plated on the bottom of a Transwell insert previously filled with Matrigel ECM protein, then allowed to invade "upwards" through the filter and into the Matrigel. Reconstruction of z-series image stacks, obtained by confocal imaging, into three-dimensional representations allows for visualization of collectively invading strands and analysis of the representation of fluorescently-labeled cells in leading versus following positions.

Models of tumor cell invasion into three-dimensional extracellular matrix better reflect the in vivo situation than two-dimensional motility assays. Using matrix invasion assays combined with confocal imaging of fluorescently-labeled cells, detailed information on invasion modes and the distinct contributions of leading versus following cells can be obtained.

モデリングとイメージング、3次元の集団細胞浸潤

A defining characteristic of cancer malignancy is invasion and metastasis 1. In some cancers (e.g. glioma 2), local invasion into surrounding healthy ...

Protocols for Assessing Radiofrequency Interactions with Gold Nanoparticles and Biological Systems for Non-invasive Hyperthermia Cancer Therapy

Cancer therapies which are less toxic and invasive than their existing counterparts are highly desirable. The use of RF electric-fields that penetrate deep into the body, causing minimal toxicity, are currently being studied as a viable means of non-invasive cancer therapy. It is envisioned that the interactions of RF energy with internalized nanoparticles (NPs) can liberate heat which can then cause overheating (hyperthermia) of the cell, ultimately ending in cell necrosis.
In the case of non-biological systems, we present detailed protocols relating to quantifying the heat liberated by highly-concentrated NP colloids. For biological systems, in the case of in vitro experiments, we describe the techniques and conditions which must be adhered to in order to effectively expose cancer cells to RF energy without bulk media heating artifacts significantly obscuring the data. Finally, we give a detailed methodology for in vivo mouse models with ectopic hepatic cancer tumors.

We describe the protocols used to investigate the interactions of 13.56 MHz radiofrequency (RF) electric-fields with gold nanoparticle colloids in both non-biological and biological systems (in vitro/vivo). These interactions are being investigated for applications in cancer therapy.

非侵襲ハイパーサーミアがん治療のための金ナノ粒子と生体系とラジオ波の相互作用を評価するためのプロトコル

Cancer therapies  which are less toxic and invasive than their existing counterparts are highly  desirable. The use of RF electric-fields that penetrate ...

Cerenkov Luminescence Imaging of Interscapular Brown Adipose Tissue

Brown adipose tissue (BAT), widely known as a &#8220;good fat&#8221; plays pivotal roles for thermogenesis in mammals. This special tissue is closely related to metabolism and energy expenditure, and its dysfunction is one important contributor for obesity and diabetes. Contrary to previous belief, recent PET/CT imaging studies indicated the BAT depots are still present in human adults. PET imaging clearly shows that BAT has considerably high uptake of 18F-FDG under certain conditions. In this video report, we demonstrate that Cerenkov luminescence imaging (CLI) with 18F-FDG can be used to optically image BAT in small animals. BAT activation is observed after intraperitoneal injection of norepinephrine (NE) and cold treatment, and depression of BAT is induced by long anesthesia. Using multiple-filter Cerenkov luminescence imaging, spectral unmixing and 3D imaging reconstruction are demonstrated. Our results suggest that CLI with 18F-FDG is a practical technique for imaging BAT in small animals, and this technique can be used as a cheap, fast, and alternative imaging tool for BAT research.

In this video report, we show the application of Cerenkov Luminescence Imaging (CLI) for interscapular brown adipose tissue in mice under activated and depressed conditions.

間褐色脂肪組織のチェレンコフ発光イメージング

Brown adipose tissue (BAT), widely known as a &#8220;good fat&#8221; plays pivotal roles for thermogenesis in mammals. This special tissue is closely ...

Murine Model for Non-invasive Imaging to Detect and Monitor Ovarian Cancer Recurrence

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of care consisting of surgical debulking and combination chemotherapy consisting of platinum and taxane compounds, almost 90% of patients recur within a few years. In these patients the development of chemoresistant disease limits the efficacy of currently available chemotherapy agents and therefore contributes to the high mortality. To discover novel therapy options that can target recurrent disease, appropriate animal models that closely mimic the clinical profile of patients with recurrent ovarian cancer are required. The challenge in monitoring intra-peritoneal (i.p.) disease limits the use of i.p. models and thus most xenografts are established subcutaneously. We have developed a sensitive optical imaging platform that allows the detection and anatomical location of i.p. tumor mass. The platform includes the use of optical reporters that extend from the visible light range to near infrared, which in combination with 2-dimensional X-ray co-registration can provide anatomical location of molecular signals. Detection is significantly improved by the use of a rotation system that drives the animal to multiple angular positions for 360 degree imaging, allowing the identification of tumors that are not visible in single orientation. This platform provides a unique model to non-invasively monitor tumor growth and evaluate the efficacy of new therapies for the prevention or treatment of recurrent ovarian cancer.

We describe a non-invasive animal imaging platform that allows the detection, quantification, and monitoring of ovarian cancer growth and recurrence. This intra-peritoneal xenograft model mimics the clinical profile of patients with ovarian cancer.

卵巣がん再発を検出し、モニターするための非侵襲的イメージングのためのマウスモデル

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of ...

Dynamic Lung Tumor Tracking for Stereotactic Ablative Body Radiation Therapy

Physicians considering stereotactic ablative body radiation therapy (SBRT) for the treatment of extracranial cancer targets must be aware of the sizeable risks for normal tissue injury and the hazards of physical tumor miss. A first-of-its-kind SBRT platform achieves high-precision ablative radiation treatment through a combination of versatile real-time imaging solutions and sophisticated tumor tracking capabilities. It uses dual-diagnostic kV x-ray units for stereoscopic open-loop feedback of cancer target intrafraction movement occurring as a consequence of respiratory motions and heartbeat. Image-guided feedback drives a gimbaled radiation accelerator (maximum 15 x 15 cm field size) capable of real-time &#177;4 cm pan-and-tilt action. Robot-driven &#177;60&#176; pivots of an integrated &#177;185&#176; rotational gantry allow for coplanar and non-coplanar accelerator beam set-up angles, ultimately permitting unique treatment degrees of freedom. State-of-the-art software aids real-time six dimensional positioning, ensuring irradiation of cancer targets with sub-millimeter accuracy (0.4 mm at isocenter). Use of these features enables treating physicians to steer radiation dose to cancer tumor targets while simultaneously reducing radiation dose to normal tissues. By adding respiration correlated computed tomography (CT) and 2-[18F] fluoro-2-deoxy-&#7429;-glucose (18F-FDG) positron emission tomography (PET) images into the planning system for enhanced tumor target contouring, the likelihood of physical tumor miss becomes substantially less1. In this article, we describe new radiation plans for the treatment of moving lung tumors.

Stereotactic ablative body radiation therapy (SBRT) involves the precise delivery of high-dose radiation to cancer tumor targets. A novel SBRT platform offers a first-of-its-kind gimbaled radiation accelerator mounted within a pivoting O-ring gantry capable of dynamic image-guided tumor tracking. Here, we describe dynamic tumor tracking for lung targets.

定位アブレーションボディ放射線治療のための動的肺腫瘍の追跡

Physicians considering stereotactic ablative body radiation therapy (SBRT) for the treatment of extracranial cancer targets must be aware of the sizeable ...

An Organotypic High Throughput System for Characterization of Drug Sensitivity of Primary Multiple Myeloma Cells

In this work we describe a novel approach that combines ex vivo drug sensitivity assays and digital image analysis to estimate chemosensitivity and heterogeneity of patient-derived multiple myeloma (MM) cells. This approach consists in seeding primary MM cells freshly extracted from bone marrow aspirates into microfluidic chambers implemented in multi-well plates, each consisting of a reconstruction of the bone marrow microenvironment, including extracellular matrix (collagen or basement membrane matrix) and stroma (patient-derived mesenchymal stem cells) or human-derived endothelial cells (HUVECs). The chambers are drugged with different agents and concentrations, and are imaged sequentially for 96 hr through bright field microscopy, in a motorized microscope equipped with a digital camera. Digital image analysis software detects live and dead cells from presence or absence of membrane motion, and generates curves of change in viability as a function of drug concentration and exposure time. We use a computational model to determine the parameters of chemosensitivity of the tumor population to each drug, as well as the number of sub-populations present as a measure of tumor heterogeneity. These patient-tailored models can then be used to simulate therapeutic regimens and estimate clinical response.

We describe a high-throughput drug sensitivity assay for primary multiple myeloma cells. It consists of a reconstruction of the bone marrow microenvironment (including extracellular matrix and stroma) in multi-well plates, and a non-invasive method for longitudinal quantification of cell viability.

一次多発性骨髄腫細胞の薬剤感受性の特徴付けのための器官のハイスループットシステム

In this work we describe a novel approach that combines ex vivo drug sensitivity assays and digital image analysis to estimate chemosensitivity and ...

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.

直接脳血栓塞栓を可視化するための複合近赤外蛍光イメージングおよびマイクロコンピュータ断層撮影

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

Quantitative Visualization of Leukocyte Infiltrate in a Murine Model of Fulminant Myocarditis by Light Sheet Microscopy

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently labelled structures in large biological specimens, and is down to cellular resolution. Meanwhile, the constant refinement of underlying protocols and the enhanced availability of specialized commercial systems enable us to investigate the microstructure of whole mouse organs and even allow for the characterization of cellular behavior in various live-cell imaging approaches. Here, we describe a protocol for the spatial whole-mount visualization and quantification of the CD45+ leukocyte population in inflamed mouse hearts. The method employs a transgenic mouse strain (CD11c.DTR)that has recently been shown to serve as a robust, inducible model for the study of the development of fulminant fatal myocarditis, characterized by lethal cardiac arrhythmias. This protocol includes myocarditis induction, intravital antibody-mediated cell staining, organ preparation, and LSFM with subsequent computer-assisted image post-processing. Although presented as a highly-adapted method for our particular scientific question, the protocol represents the blueprint of an easily adjustable system that can also target completely different fluorescent structures in other organs and even in other species.

Here, we describe a light-sheet microscopy approach to visualize the cardiac CD45+ leukocyte infiltrate in a murine model of aseptic fulminant myocarditis, which is induced by the intratracheal diphtheria toxin treatment of CD11c.DTR mice.

光シート顕微鏡による劇症心筋炎のマウスモデルにおける白血球浸潤の定量的可視化

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently ...

Cardiac Magnetic Resonance for the Evaluation of Suspected Cardiac Thrombus: Conventional and Emerging Techniques

We present the conventional cardiac magnetic resonance (CMR) protocol for evaluating a suspected thrombus and highlight emerging techniques.&#160;The appearance of a mass on certain magnetic resonance (MR) sequences can help differentiate a thrombus from competing diagnoses such as a tumor. T1 and T2 signal characteristics of a thrombus are related to the evolution of hemoglobin properties. A thrombus typically does not enhance following contrast administration, which also helps differentiation from a tumor. We also highlight the emerging role of T1 mapping in the evaluation of a thrombus, which can add another level of support in diagnosis. Prior to any CMR exam, patient screening and interviews are critical to ensure safety and to optimize patient comfort. Effective communication during the exam between the technologist and the patient promotes proper breath holding technique and higher quality images. Volumetric post processing and structured reporting are helpful to ensure that the radiologist answers the ordering services&#39; question and communicates these results effectively. Optimal pre-MR safety evaluation, CMR exam execution, and post exam processing and reporting allow for delivery of high quality radiological service in the evaluation of a suspected cardiac thrombus.

The goal of this article is to describe how cardiac magnetic resonance can be used for the evaluation and diagnosis of a suspected cardiac thrombus. The method presented will describe data acquisition as well as the pre-procedure and post-procedure protocol.

疑わしい心臓血栓の評価のための心臓磁気共鳴:従来技術と新興技術

We present the conventional cardiac magnetic resonance (CMR) protocol for evaluating a suspected thrombus and highlight emerging techniques.&#160;The ...