This work is funded by the Cure for Cancer Foundation, Dutch Technology Foundation (STW) and The Netherlands Organisation for Health Research and Development (ZonMw).

提示手順は、学術医療センターアムステルダム、登録番号NL41985.018の治験審査委員会によって承認された研究プロトコルの下で行われます。書面によるインフォームドコンセントは、すべての参加者から要求されている。 1.システム<ol><li>この実験のために、1,280-1,350波長帯13で動作する、フーリエドメインOCTシステムを使用する。フーリエ領域低コヒーレンス干渉法は、第一世代の時間領域OCTシステムと比較した場合、データ収集速度を増加させる連続スキャンを可能にする。注：OCTシステムは、約90°の角度で螺旋状に走査し、光ファイバプローブとインターフェースされる。これは、2.7Fの外径（0.9 mm）の135センチ挿入可能な長さを有している。プローブは、54ミリメートルのプルバック範囲で駆動モータと光コントローラ（取付ドック）を通じて10月コンソールに接続します。取得したOCTデータセットは、541の断面像（Bスキャ​​ン）ワットから成り15μmの距離分解能i番目（ 図1,2）。 </li><li> Kodach ら 14,15により、前述のように正確で再現性の減衰測定を保証するために、脂肪乳剤、（ 例えば 、イントラリピッド）の重量百分率に基づいてμ10月増加する濃度について測定することによって較正する。 要するに： <ol><li> 0.125、0.250、0.5、1.0、2.0、4.0、10、15、20（ストック）パーセントの濃度を達成するために、脱イオンH 2 O、20％の脂肪乳剤の標準的なバッチを希釈する。 <ol><li>脂肪乳剤の混合物200mlにOCTプローブを配置し、OCT計測を取得する。 </li><li>相互参照は、文献中で既知の値とμ10月の値を抽出した。 </li></ol></li></ol></li></ol> 2.タイム·アウトと患者位置決め<ol><li> PR、手順を開始する前に、名前、生年月日、手順を確認し、「タイムアウト」を実行ocedural側、抗凝固剤の使用、およびアレルギー。 </li><li>腫瘍の位置に応じて、どちらを起こしやすいまたは側臥位で患者を置く。十分な支援を患者に提供し、彼/彼女は20から40分間にわたってこの位置に快適であることを期待しているかどうかを確認。 </li><li>超音波（US）16を使用して、腫瘍をローカライズし、永久的なインクで皮膚に針落ち点をマーク。 注：コンピュータ断層撮影（CT）を使用する場合は、アクセス針の好ましい位置をローカライズするために柔軟な針誘導テンプレートを使用します。 </li></ol> 3.消毒と滅菌ドレープ<ol><li>手術帽と口カバーに入れてください。 </li><li>以前に配置された針落ちマーク（ステップ2.3）を削除しないように注意しながら、クロルヘキシジン/アルコール溶液を用いて、穿刺部位の周りの皮膚を清掃してください。広域消毒すると、予期しないアクセス針の再配置の場合の追加の洗浄の必要性を防ぐことができます。 </li><li> Rはた10mlシリンジ、鈍吸引針、21 G注射針、メス、15 G同軸導入針、18 Gのトロカール針、及び16：無菌コンテンツのegardを含む経皮的穿刺セットを開きGコア生検銃。 </li><li>その後手の消毒剤を適用して、徹底的に手を洗う。手術衣及び滅菌手袋を着用してください。 </li><li>無菌ドレープで患者をカバーしています。 </li><li>超音波プローブの周りに無菌カバーを適用し、所定の位置にニードルガイドを修正。 </li></ol> 4. 10月の準備<ol><li> 10月コンソールを起動し、コンソールインターフェイスを使用して、患者ID、姓、名、およびDOB（生年月日）をラベルの分野で患者の詳細を入力してください。 </li><li>無菌のコンテンツのに関しては、OCTプローブ、無菌の取り付けドックカバー、および5ミリリットルルアーロックシリンジを含む10月のパッケージを展開。 </li><li> 10月コンソールマウントドックに無菌のカバーを適用します。非無菌取り付けドックを導くことが必要ですアシスタントの助けを。 </li><li> 0.9％NaClで5ミリリットル注射器を記入し、フラッシングポートに取り付けます。水がプローブカバーの先端部に表示されるまで、OCTプローブをフラッシュします。 </li><li>取り付けドックにOCTプローブをロードします。回転させて、適切な機能を確認した赤色の光を放出するプローブをロードした後。損傷のリスクを最小限に抑えるために紅潮し、ロード中にその保護カバーにプローブのままにしておきます。 </li><li>そのカバーからOCTプローブを削除します。硬い表面にプローブを配置し、チップを短縮するメスを使用しています。光ファイバとプリズムに圧力を最小限にするために、切削中にプローブの先端部を固定します。オリエンテーションのために放出される（赤色）光を用いて、プリズムから遠位5ミリメートルをカット。 </li></ol> 5.穿刺<ol><li> 2％リドカインた（20mg / ml）を用いて、皮膚および深層を麻酔。リドカインを有効にするためにできるように、数分待ちます。痛みがある場合に患者情報を確認して下さい。 </li><li>ニードルガイドを使用して、配置する撮影により位置を確認する15 G同軸導入針。配置が満足のいくものである場合には、オブチュレータ（鋭い針芯）を削除。 </li><li>腫瘍を貫通、導入針を通じて18のGトロカール針を配置します。再びイメージングと針の位置を確認します。配置が満足のいくものである場合には、閉塞具を取り外します。 </li><li>抵抗を感じまで、トロカール針までのOCTプローブフィード。 </li><li> OCTプローブを固定して、腫瘍組織をOCTプローブを露出する、トロカール針を後退させる。腫瘍内のトロカール針の先端を維持することは、呼吸サイクル中のOCTプローブのよじれを最小限に抑えます。これは、プローブの損傷のリスクを低下させる。 </li><li> 10月スキャン： <ol><li>データセットあたり541 Bスキャンに設定コンソールで、10月のスキャンを実行します。ここで使用されるOCTシステムは、特定のパラメータ調整を必要としない5.4センチメートルの長さにわたって、自動プルバックを実行します。 </li><li> （品質、工芸品のためのスキャンおよび固形組織の外観を確認してください図1A）。アーティファクトは最も一般的に円形のバンドが通常の10月パターン（ 図1B）から出て立って現れる。 </li><li>人工物が再スキャン後に続く場合はプローブを交換してください。 </li></ol></li><li> 10月3日のデータセットの最小までの手順を繰り返し5.6が取得される。 </li><li>インプレース導入針を残して、OCTプローブ及びトロカール針を外します。 </li><li>コア生検銃をARMおよびイメージング上の位置を確認し、導入針を介して配置します。 </li><li>位置決めが満足のいくものである場合には、生検銃を発射。 </li><li>病理部門プロトコルに従って容器に生検材料を配置します。ここでは、紙のインレイとペトリ皿、上の場所を検は十分に0.9％NaClで飽和。 </li><li>コア生検の品質を確認し、十分な材料が得られるまでステップ5.9と5.10を繰り返します。 </li></ol>

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The authors of this article have nothing to disclose.

この公報では、腎臓の10​​月、ベースの経皮的、針の実現可能性について報告する。これは、「光学的生検」と呼ば腫瘍分化のための臨床的に適用可能な技術、に10月の発展に不可欠な最初のステップです。私たちの最初の25人の患者が簡単で安全手順であることが、経皮10月を示している。光学的生検は、従来のコア生検上の2つの利点がある。まず、OCTデータのリアルタイム取得および分?...

過去数十年、腎大衆1,2の発生率の着実な増加を示している。今まで、腎腫治療の決定は、主にMRIとCT撮像特性、年齢および併存疾患に基づいて行われてきた。しかしこれらの診断法と臨床パラメーターは本当に腎質量の悪性の可能性を検出するためのフィネスを欠いている。病理学的評価のための十分な組織（診断）とコア生検または細針吸引95〜100％3の範囲の感度と特異性の両方を持つ客観的な腫瘍の分化を提供します。そのため生検は不審な腎大衆4,5の評価で受け入れを集めています。しかし、十分な組織生検なしでは、遅延、診断、または腎実質（非診断）法線が全体の10〜20％の割合で発生し、小さな腎腫瘤では30％までで（&lt;4センチのSRM）を確立する追加のための頻繁な必要性に起因する診断プロセス生検手順3,5。 光コヒーレンストモグラフィ（OCT）は、腎腫分化における上記の障害を克服する可能性を有する新規なイメージングモダリティである。近赤外光の後方散乱に基づいて、10月2~3程度の効果的な組織浸透に15μmの距離分解能を有する画像を提供する（ 図1,2）。ファーバーらによって記載されるように6：組織浸透のミリメートル当たり信号強度の損失、組織特異的な光散乱の結果は、減衰係数（ミリメートル-1μOCT）として表される。組織学的特徴は、組織分化の定量的パラメータ（ 図3）を提供するのOCT値をμと相関することができる。 発癌中は、悪性細胞が増加した数、とのより大きく、より不規則な形の核を表示<html>高い屈折率およびより活性なミトコンドリア。による細胞成分のこの過剰発現、μ10月の変化が良性腫瘍または非罹患組織7に悪性腫瘍を比較する際に予想される。 最近では、良性と悪性の腎腫瘤8,9を区別する表面的な10月の能力を研究した。 16人の患者では、腫瘍組織の術中のOCT測定は、外部に配置されたOCTプローブを用いて得た。同じ患者の非罹患組織のOCT測定値からなる制御アーム。正常組織は、腫瘍の分化のために、OCTの可能性を確認し、悪性組織と比較して有意に低い中央値の減衰係数を示した。この定量分析は、尿路上皮癌10,11及び外陰上皮新形成分化12などの悪性組織のグレード他のタイプと同様の方法で適用されている。 と診断された患者でENT &quot;&gt;我々は、現場での腫瘍分化と組み合わせたリアルタイムイメージングを提供し、光学的生検に10月を開発することを目指しています。現在の研究の目的は、経皮、針ベースを記述することで、10月のアプローチ腎臓の質量を強化する固体。この方法の説明には、我々の知る限り、腎腫瘍の10月に基づく針の可能性を評価する最初のものです。

<table><tbody><tr><td>15 G/7.5 cm Co-Axial Introducer Needle</td><td>Angiotech, Gainesville, USA</td><td>MCXS1612SX</td><td></td></tr><tr><td>18 G/20 cm Trocar Needle</td><td>Cook medical, Bloomington, USA</td><td>DTN-18-20.0-U</td><td></td></tr><tr><td>16 G/20 cm Quick-Core Biopsy Gun</td><td>Cook Medical, Bloomington, USA</td><td>G07827</td><td></td></tr><tr><td>Ilumien Optis PCI Optimization System (OCT &amp;amp; FFR)</td><td>St. Jude medical, St. Paul, USA</td><td>C408650</td><td>Part of Dragonfly Kit.&lt;br/&gt; St. Jude medical, St. Paul, USA. (C4088643)</td></tr><tr><td>Dragonfly Duo Imaging Catheter</td><td>LightLab Imaging, Westford, USA</td><td>C408644</td><td>Part of Dragonfly Kit.&lt;br/&gt; St. Jude medical, St. Paul, USA. (C4088643)</td></tr><tr><td>Sterile Dock Cover</td><td>CFI Med. Solutions, Fenton, USA</td><td>200-700-00</td><td>Part of Dragonfly Kit.&lt;br/&gt; St. Jude medical, St. Paul, USA. (C4088643)</td></tr><tr><td>5 ml Luer-lock Syringe</td><td>Merit Med. Syst., South Jordan, USA</td><td>C408647</td><td></td></tr><tr><td>10 ml Syringe</td><td>BD, Franklin Lakes, USA</td><td>300912</td><td></td></tr><tr><td>18 G Blunt Fill Needle</td><td>BD, Franklin Lakes, USA</td><td>305180</td><td></td></tr><tr><td>21 G Injection Needle</td><td>BD, Franklin Lakes, USA</td><td>301155</td><td></td></tr><tr><td>Sterile scalpel</td><td>BD, Franklin Lakes, USA</td><td>372611</td><td></td></tr><tr><td>NaCl 0,9% solution</td><td>Braun, Melsungen AG, Germany</td><td>222434</td><td></td></tr><tr><td>Lidoca&amp;iuml;ne HCl 2% (20 mg/ml) solution</td><td>Braun, Melsungen AG, Germany</td><td>3624480</td><td></td></tr><tr><td>Sterile Ultrasound Gel, Aquasonic 100</td><td>Parker Lab. Inc., Fairfield, USA</td><td>GE424609</td><td></td></tr><tr><td>Sterile Ultrasound Cover</td><td>Microtek Med., Alpharetta, USA</td><td>PC1289EU</td><td></td></tr><tr><td>Pathology Container</td><td></td><td></td><td></td></tr><tr><td>AMIRA software package</td><td>FEI Visualization Sciences Group, Hillsboro, USA</td><td>Software platform for 3D data analysis</td><td></td></tr><tr><td>FIJI software package (open source)</td><td>Open source, http://fiji.sc/Fiji</td><td></td><td>Open source image processing software</td></tr></tbody></table>

in vivo, percutaneous, needle based, optical coherence tomography of renal masses

Optical coherence tomography (OCT) is the optical equivalent of ultrasound imaging, based on the backscattering of near infrared light. OCT provides real time images with a 15 &#181;m axial resolution at an effective tissue penetration of 2-3 mm. Within the OCT images the loss of signal intensity per millimeter of tissue penetration, the attenuation coefficient, is calculated. The attenuation coefficient is a tissue specific property, providing a quantitative parameter for tissue differentiation. 
Until now, renal mass treatment decisions have been made primarily on the basis of MRI and CT imaging characteristics, age and comorbidity. However these parameters and diagnostic methods lack the finesse to truly detect the malignant potential of a renal mass. A successful core biopsy or fine needle aspiration provides objective tumor differentiation with both sensitivity and specificity in the range of 95-100%. However, a non-diagnostic rate of 10-20% overall, and even up to 30% in SRMs, is to be expected, delaying the diagnostic process due to the frequent necessity for additional biopsy procedures. 
We aim to develop OCT into an optical biopsy, providing real-time imaging combined with on-the-spot tumor differentiation. This publication provides a detailed step-by-step approach for percutaneous, needle based, OCT of renal masses.

最初の25の腫瘍（23例）のうち、24成功したOCTの手順の合計を行った。あるケースでは、プローブの故障は、OCTスキャンを取得することができないことにつながった。二つの有害事象（AE）は、議論の項で詳細に記載されており、発生した。一般的な患者の特徴を表1に記載されている。 10月コンソールは、取得したデータセットの即時定性分析のために、リア...

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In Vivo, Percutaneous, Needle Based, Optical Coherence Tomography of Renal Masses

Optical coherence tomography (OCT) is a high resolution imaging technique that allows analysis of tissue specific optical properties providing the means for tissue differentiation. We developed needle based OCT, providing real-time imaging combined with on-the-spot tumor differentiation. This publication describes a method for percutaneous, needle based OCT of renal masses.

生体内 、経皮、ニードルベース、腎大衆の光コヒーレンストモグラフィーで

Optical coherence tomography (OCT) is the optical equivalent of ultrasound imaging, based on the backscattering of near infrared light. OCT provides real ...

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Research

JoVE Journal

Medicine

In Vivo, Percutaneous, Needle Based, Optical Coherence Tomography of Renal Masses

8.8K ビュー.  Academic Medical Center. Optical coherence tomography (OCT) is a high resolution imaging technique that allows analysis of tissue specific optical properties providing the means for tissue differentiation. We developed needle based OCT, providing real-time imaging combined with on-the-spot tumor differentiation. This publication describes a method for percutaneous, needle based OCT of renal masses.

ビデオ: In Vivo, Percutaneous, Needle Based, Optical Coherence Tomography of Renal Masses

Optical Frequency Domain Imaging of Ex vivo Pulmonary Resection Specimens: Obtaining One to One Image to Histopathology Correlation

Lung cancer is the leading cause of cancer-related deaths1. Squamous cell and small cell cancers typically arise in association with the conducting airways, whereas adenocarcinomas are typically more peripheral in location. Lung malignancy detection early in the disease process may be difficult due to several limitations: radiological resolution, bronchoscopic limitations in evaluating tissue underlying the airway mucosa and identifying early pathologic changes, and small sample size and/or incomplete sampling in histology biopsies. High resolution imaging modalities, such as optical frequency domain imaging (OFDI), provide non-destructive, large area 3-dimensional views of tissue microstructure to depths approaching 2 mm in real time (Figure 1)2-6. OFDI has been utilized in a variety of applications, including evaluation of coronary artery atherosclerosis6,7 and esophageal intestinal metaplasia and dysplasia6,8-10.
Bronchoscopic OCT/OFDI has been demonstrated as a safe in vivo imaging tool for evaluating the pulmonary airways11-23 (Animation). OCT has been assessed in pulmonary airways16,23 and parenchyma17,22 of animal models and in vivo human airway14,15. OCT imaging of normal airway has demonstrated visualization of airway layering and alveolar attachments, and evaluation of dysplastic lesions has been found useful in distinguishing grades of dysplasia in the bronchial mucosa11,12,20,21. OFDI imaging of bronchial mucosa has been demonstrated in a short bronchial segment (0.8 cm)18. Additionally, volumetric OFDI spanning multiple airway generations in swine and human pulmonary airways in vivo has been described19. Endobronchial OCT/OFDI is typically performed using thin, flexible catheters, which are compatible with standard bronchoscopic access ports. Additionally, OCT and OFDI needle-based probes have recently been developed, which may be used to image regions of the lung beyond the airway wall or pleural surface17.
While OCT/OFDI has been utilized and demonstrated as feasible for in vivo pulmonary imaging, no studies with precisely matched one-to-one OFDI:histology have been performed. Therefore, specific imaging criteria for various pulmonary pathologies have yet to be developed. Histopathological counterparts obtained in vivo consist of only small biopsy fragments, which are difficult to correlate with large OFDI datasets. Additionally, they do not provide the comprehensive histology needed for registration with large volume OFDI. As a result, specific imaging features of pulmonary pathology cannot be developed in the in vivo setting. Precisely matched, one-to-one OFDI and histology correlation is vital to accurately evaluate features seen in OFDI against histology as a gold standard in order to derive specific image interpretation criteria for pulmonary neoplasms and other pulmonary pathologies. Once specific imaging criteria have been developed and validated ex vivo with matched one-to-one histology, the criteria may then be applied to in vivo imaging studies. Here, we present a method for precise, one to one correlation between high resolution optical imaging and histology in ex vivo lung resection specimens. Throughout this manuscript, we describe the techniques used to match OFDI images to histology. However, this method is not specific to OFDI and can be used to obtain histology-registered images for any optical imaging technique. We performed airway centered OFDI with a specialized custom built bronchoscopic 2.4 French (0.8 mm diameter) catheter. Tissue samples were marked with tissue dye, visible in both OFDI and histology. Careful orientation procedures were used to precisely correlate imaging and histological sampling locations. The techniques outlined in this manuscript were used to conduct the first demonstration of volumetric OFDI with precise correlation to tissue-based diagnosis for evaluating pulmonary pathology24. This straightforward, effective technique may be extended to other tissue types to provide precise imaging to histology correlation needed to determine fine imaging features of both normal and diseased tissues.

Optical Frequency Domain Imaging of Ex vivo Pulmonary Resection Specimens: Obtaining One to One Image to Histopathology Correlation

A method to image ex vivo pulmonary resection specimens with optical frequency domain imaging (OFDI) and obtain precise correlation to histology is described, which is essential to developing specific OFDI interpretation criteria for pulmonary pathology. This method is applicable to other tissue types and imaging techniques to obtain precise imaging to histology correlation for accurate image interpretation and assessment. Imaging criteria established with this technique would then be applicable to image assessment in future in vivo studies.

の光周波数領域イメージング<em&gt;エクスビボ</em&gt;肺切除標本：組織病理相関へのOne to Oneイメージの入手

Lung cancer is the leading cause of cancer-related deaths1. Squamous cell and small cell cancers typically arise in association with the conducting ...

生物工学

Longitudinal Morphological and Physiological Monitoring of Three-dimensional Tumor Spheroids Using Optical Coherence Tomography

Tumor spheroids have been developed as a three-dimensional (3D) cell culture model in cancer research and anti-cancer drug discovery. However, currently, high-throughput imaging modalities utilizing bright field or fluorescence detection, are unable to resolve the overall 3D structure of the tumor spheroid due to limited light penetration, diffusion of fluorescent dyes and depth-resolvability. Recently, our lab demonstrated the use of optical coherence tomography (OCT), a label-free and non-destructive 3D imaging modality, to perform longitudinal characterization of multicellular tumor spheroids in a 96-well plate. OCT was capable of obtaining 3D morphological and physiological information of tumor spheroids growing up to about 600 &#181;m in height. In this article, we demonstrate a high-throughput OCT (HT-OCT) imaging system that scans the whole multi-well plate and obtains 3D OCT data of tumor spheroids automatically. We describe the details of the HT-OCT system and construction guidelines in the protocol. From the 3D OCT data, one can visualize the overall structure of the spheroid with 3D rendered and orthogonal slices, characterize the longitudinal growth curve of the tumor spheroid based on the morphological information of size and volume, and monitor the growth of the dead-cell regions in the tumor spheroid based on optical intrinsic attenuation contrast. We show that HT-OCT can be used as a high-throughput imaging modality for drug screening as well as characterizing biofabricated samples.

Optical coherence tomography (OCT), a three-dimensional imaging technology, was used to monitor and characterize the growth kinetics of multicellular tumor spheroids. Precise volumetric quantification of tumor spheroids using a voxel counting approach, and label-free dead tissue detection in the spheroids based on intrinsic optical attenuation contrast, were demonstrated.

光コヒーレンストモグラフィーを用いた三次元腫瘍回転楕円体の縦断的形態学的および生理学的監視

Tumor spheroids have been developed as a three-dimensional (3D) cell culture model in cancer research and anti-cancer drug discovery. However, currently, ...

がん研究

前眼部HR-OCTを用いた眼表面扁平上皮腫瘍の評価

Anterior High-Resolution Optical Coherence Tomography in the Diagnosis and Therapeutic Monitoring of Ocular Surface Squamous Neoplasia

Ocular surface squamous neoplasia (OSSN) is the most common tumor of the ocular surface, ranging from mild dysplasia to invasive squamous carcinoma. Traditionally, the diagnosis of OSSN relies on histopathological confirmation followed by a full-thickness biopsy. However, in the past two decades, the therapeutic approach to OSSN has shifted from surgical intervention to topical chemotherapy regimens in clinical settings. This shift emphasizes the need for less invasive or non-invasive methods to diagnose ocular surface pathologies. Among various imaging devices, commercially available high-resolution optical coherence tomography (HR-OCT) has emerged as a powerful tool for the characterization of OSSN. HR-OCT provides an in vivo, cross-sectional view of ocular surface lesions, offering an "optical biopsy" for OSSN with high sensitivity and specificity. It provides valuable information in differentiating intraepithelial or invasive OSSN from other benign lesions. Additionally, HR-OCT can be used to monitor the response to topical chemotherapy and to detect subclinical OSSN during follow-up visits. In this article, the scanning protocol for image acquisition is presented, and image interpretation for OSSN is outlined. This standardized, practical, and reproducible approach is recommended in clinical workflows and is expected to assist clinicians in the management of OSSN.

著者スポットライト:眼表面扁平上皮腫瘍を特徴付けるための非侵襲的ツールとしての前方HR-OCT

Anterior segment high-resolution optical coherence tomography (HR-OCT) is a promising non-invasive modality for the diagnosis and therapeutic evaluation of ocular surface squamous neoplasia (OSSN). Here, the system setup, scanning technique, and representative diagnostic results are presented.

眼表面扁平上皮腫瘍の診断と治療モニタリングにおける前方高解像度光干渉断層撮影法

Ocular surface squamous neoplasia (OSSN) is the most common tumor of the ocular surface, ranging from mild dysplasia to invasive squamous carcinoma. ...

工学

Optical Coherence Tomography: Imaging Mouse Retinal Ganglion Cells In Vivo

Structural changes in the retina are common manifestations of ophthalmic diseases. Optical coherence tomography (OCT) enables their identification in vivo&#8212;rapidly, repetitively, and at a high resolution. This protocol describes OCT imaging in the mouse retina as a powerful tool to study optic neuropathies (OPN). The OCT system is an interferometry-based, non-invasive alternative to common post mortem histological assays. It provides a fast and accurate assessment of retinal thickness, allowing the possibility to track changes, such as retinal thinning or thickening. We present the imaging process and analysis with the example of the Opa1delTTAG mouse line. Three types of scans are proposed, with two quantification methods: standard and homemade calipers. The latter is best for use on the peripapillary retina during radial scans; being more precise, is preferable for analyzing thinner structures. All approaches described here are designed for retinal ganglion cells (RGC) but are easily adaptable to other cell populations. In conclusion, OCT is efficient in mouse model phenotyping and has the potential to be used for the reliable evaluation of therapeutic interventions.

Optical Coherence Tomography: Imaging Mouse Retinal Ganglion Cells In Vivo

This manuscript describes a protocol for the in vivo imaging of the mouse retina with high-resolution spectral domain optical coherence tomography (SD-OCT). It focuses on retinal ganglion cells (RGC) in the peripapillary region, with several scanning and quantifying approaches described.

光干渉断層計: マウス網膜神経節細胞In Vivoイメージング

Structural changes in the retina are common manifestations of ophthalmic diseases. Optical coherence tomography (OCT) enables their identification in ...

神経科学

In Vivo Multimodal Imaging and Analysis of Mouse Laser-Induced Choroidal Neovascularization Model

Laser-induced choroidal neovascularization (CNV) is a well-established model to mimic the wet form of age-related macular degeneration (AMD). In this protocol, we aim to guide the reader not simply through the technical considerations of generating laser-induced lesions to trigger neovascular processes, but rather focus on the powerful information that can be obtained from multimodal longitudinal in vivo imaging throughout the follow-up period.
The laser-induced mouse CNV model was generated by a diode laser administration. Multimodal in vivo imaging techniques were used to monitor CNV induction, progression and regression. First, spectral domain optical coherence tomography (SD-OCT) was performed immediately after the lasering to verify a break of Bruch&#39;s membrane. Subsequent in vivo imaging using fluorescein angiography (FA) confirmed successful damage of Bruch&#39;s membrane from serial images acquired at the choroidal level. Longitudinal follow-up of CNV proliferation and regression on days 5, 10, and 14 after the lasering was performed using both SD-OCT and FA. Simple and reliable grading of leaky CNV leasions from FA images is&#160;presented. Automated segmentation for measurement of total retinal thickness, combined with manual caliber application for measurement of retinal thickness at CNV sites, allow unbiased evaluation of the presence of edema. Finally, histological verification of CNV is performed using isolectin GS-IB4 staining on choroidal flatmounts. The staining is thresholded, and the isolectin-positive area is calculated with ImageJ.
This protocol is especially useful in therapeutics studies requiring high-throughput-like screening of CNV pathology as it allows fast, multimodal, and reliable classification of CNV pathology and retinal edema. In addition, high resolution SD-OCT enables the recording of other pathological hallmarks, such as the accumulation of subretinal or intraretinal fluid. However, this method does not provide a possibility to automate CNV volume analysis from SD-OCT images, which has to be performed manually.

In Vivo Multimodal Imaging and Analysis of Mouse Laser-Induced Choroidal Neovascularization Model

Here, we present the usefulness of longitudinal in vivo imaging in the follow-up of morphological changes of laser-induced choroidal neovascularization in mice.

生体内でマルチ モーダル イメージングとマウス レーザー誘発脈絡膜新生血管モデルの解析

Laser-induced choroidal neovascularization (CNV) is a well-established model to mimic the wet form of age-related macular degeneration (AMD). In this ...

In vivo Structural Assessments of Ocular Disease in Rodent Models using Optical Coherence Tomography

Spectral-domain optical coherence tomography (SD-OCT) is useful for visualizing retinal and ocular structures in vivo. In research, SD-OCT is a valuable tool to evaluate and characterize changes in a variety of retinal and ocular disease and injury models. In light induced retinal degeneration models, SD-OCT can be used to track thinning of the photoreceptor layer over time. In glaucoma models, SD-OCT can be used to monitor decreased retinal nerve fiber layer and total retinal thickness and to observe optic nerve cupping after inducing ocular hypertension. In diabetic rodents, SD-OCT has helped researchers observe decreased total retinal thickness as well as decreased thickness of specific retinal layers, particularly the retinal nerve fiber layer with disease progression. In mouse models of myopia, SD-OCT can be used to evaluate axial parameters, such as axial length changes. Advantages of SD-OCT include in vivo imaging of ocular structures, the ability to quantitatively track changes in ocular dimensions over time, and its rapid scanning speed and high resolution. Here, we detail the methods of SD-OCT and show examples of its use in our laboratory in models of retinal degeneration, glaucoma, diabetic retinopathy, and myopia. Methods include anesthesia, SD-OCT imaging, and processing of the images for thickness measurements.

Here, we describe the use of spectral-domain optical coherence tomography (SD-OCT) to visualize retinal and ocular structures in vivo in models of retinal degeneration, glaucoma, diabetic retinopathy, and myopia.

光コヒーレンストモグラフィーを用いたげっ歯類モデルにおける眼疾患のin vivo構造評価

Spectral-domain optical coherence tomography (SD-OCT) is useful for visualizing retinal and ocular structures in vivo. In research, SD-OCT is a valuable ...

Retinal Vascular Reactivity as Assessed by Optical Coherence Tomography Angiography

The vascular supply to the retina has been shown to dynamically adapt through vasoconstriction and vasodilation to accommodate the metabolic demands of the retina. This process, referred to as retinal vascular reactivity (RVR), is mediated by neurovascular coupling, which is impaired very early in retinal vascular diseases such as diabetic retinopathy. Therefore, a clinically feasible method of assessing vascular function may be of significant interest in both research and clinical settings. Recently, in vivo imaging of the retinal vasculature at the capillary level has been made possible by the FDA approval of optical coherence tomography angiography (OCTA), a noninvasive, minimal risk and dyeless angiography method with capillary level resolution. Concurrently, physiological and pathological changes in RVR have been shown by several investigators. The method shown in this manuscript is designed to investigate RVR using OCTA with no need for alterations to the clinical imaging procedures or device. It demonstrates real time imaging of the retina and retinal vasculature during exposure to hypercapnic or hyperoxic conditions. The exam is easily performed with two personnel in under 30 min with minimal subject discomfort or risk. This method is adaptable to other ophthalmic imaging devices and the applications may vary based on the composition of the gas mixture and patient population. A strength of this method is that it allows for an investigation of retinal vascular function at the capillary level in human subjects in vivo. Limitations of this method are largely those of OCTA and other retinal imaging methods including imaging artifacts and a restricted dynamic range. The results obtained from the method are OCT and OCTA images of the retina. These images are amenable to any analysis that is possible on commercially available OCT or OCTA devices. The general method, however, can be adapted to any form of ophthalmic imaging.

This article describes a method for measuring retinal vasculature reactivity in vivo with human subjects using a gas breathing provocation technique to deliver vasoactive stimuli while acquiring retinal images.

光コヘレンス断層撮影解剖学によるレチナル血管反応性

The vascular supply to the retina has been shown to dynamically adapt through vasoconstriction and vasodilation to accommodate the metabolic demands of ...

Application of Optical Coherence Tomography to a Mouse Model of Retinopathy

Optical coherence tomography (OCT) offers a noninvasive method for the diagnosis of retinopathy. The OCT machine can capture retinal crosssectional images from which the retinal thickness can be calculated. Although OCT is widely used in clinical practice, its application in basic research is not as prevalent, especially in small animals such as mice. Because of the small size of their eyeballs, it is challenging to conduct fundus imaging examinations in mice. Therefore, a specialized retinal imaging system is required to accommodate OCT imaging on small animals. This article demonstrates a small-animal-specific system for OCT examination procedures and a detailed method for image analysis. The results of retinal OCT examination of very-low-density lipoprotein receptor (Vldlr) knockout mice and C57BL/6J mice are presented. The OCT images of C57BL/6J mice showed retinal layers, while those of Vldlr knockout mice showed subretinal neovascularization and retinal thinning. In summary, OCT examination could facilitate the noninvasive detection and measurement of retinopathy in mouse models.

Here, we describe an in vivo imaging technique using optical coherence tomography to facilitate the diagnosis and quantitative measurement of retinopathy in mice.

光干渉断層撮影法の網膜症モデルマウスへの応用

Optical coherence tomography (OCT) offers a noninvasive method for the diagnosis of retinopathy. The OCT machine can capture retinal crosssectional images ...

生体内、経皮、ニードルベース、腎大衆の光コヒーレンストモグラフィーで

要約

さらに動画を探す

この動画の章

の光周波数領域イメージング

光コヒーレンストモグラフィーを用いた三次元腫瘍回転楕円体の縦断的形態学的および生理学的監視

眼表面扁平上皮腫瘍の診断と治療モニタリングにおける前方高解像度光干渉断層撮影法

眼表面扁平上皮腫瘍の診断と治療モニタリングにおける前方高解像度光干渉断層撮影法

眼表面扁平上皮腫瘍の診断と治療モニタリングにおける前方高解像度光干渉断層撮影法

光干渉断層計: マウス網膜神経節細胞In Vivoイメージング

生体内でマルチモーダルイメージングとマウスレーザー誘発脈絡膜新生血管モデルの解析

光コヒーレンストモグラフィーを用いたげっ歯類モデルにおける眼疾患のin vivo構造評価

光コヘレンス断層撮影解剖学によるレチナル血管反応性

光干渉断層撮影法の網膜症モデルマウスへの応用

生体内 、経皮、ニードルベース、腎大衆の光コヒーレンストモグラフィーで

要約

さらに動画を探す

この動画の章

の光周波数領域イメージング

光コヒーレンストモグラフィーを用いた三次元腫瘍回転楕円体の縦断的形態学的および生理学的監視

眼表面扁平上皮腫瘍の診断と治療モニタリングにおける前方高解像度光干渉断層撮影法

眼表面扁平上皮腫瘍の診断と治療モニタリングにおける前方高解像度光干渉断層撮影法

眼表面扁平上皮腫瘍の診断と治療モニタリングにおける前方高解像度光干渉断層撮影法

光干渉断層計: マウス網膜神経節細胞In Vivoイメージング

生体内でマルチ モーダル イメージングとマウス レーザー誘発脈絡膜新生血管モデルの解析

光コヒーレンストモグラフィーを用いたげっ歯類モデルにおける眼疾患のin vivo構造評価

光コヘレンス断層撮影解剖学によるレチナル血管反応性

光干渉断層撮影法の網膜症モデルマウスへの応用

生体内、経皮、ニードルベース、腎大衆の光コヒーレンストモグラフィーで

生体内でマルチモーダルイメージングとマウスレーザー誘発脈絡膜新生血管モデルの解析