Name: intravital microscopy of leukocyte-endothelial and platelet-leukocyte interactions in mesenterial veins in mice
Uploaded: 2015-08-13T13:00:00.000+00:00
Duration: 5 min 12 s
Description: Watch this Scientific Journal Video about Intravital Microscopy of Leukocyte-endothelial and Platelet-leukocyte Interactions in Mesenterial Veins in Mice at JoVE.com

This work was supported by the Deutsche Forschungsgemeinschaft (DU 1190/3-1).

全ての動物実験は、ドイツ動物保護法（TierSchG）に準拠して行きました。 FELASA（www.felasa.eu/guidelines.php）と全国動物福祉本体GV-SOLAS条約（www.gv-solas.de/index.html）で定義されているようにマウスが収容され、良好な動物の慣行に従って取り扱われました。フライブルク大学の動物福祉委員会だけでなく、自治体（Regierungspräsidiumフライブルク）は、すべての動物実験を承認しました。 1.動物の取り扱い、準備および炎症の誘導<ol><li> 16〜20グラムの重量範囲で6週齢の雄マウス - 4を使用してください。注：マウスは古いまたは重い場合、それらは顕微鏡観察を制限する、血管の周囲の過剰な脂肪を提示します。 </li><li>手術前にすべてのツールと​​顕微鏡テーブルを滅菌します。 </li><li>注入の20mg / kgのLPS（リポポリサッカライド）を腹腔内4細菌inflammを誘導するために生きているマウスに、顕微鏡の前に時間エーション。 </li><li>プラスチック容器および腸間膜組織を加湿するために37℃の水浴中で0.9％食塩水を予熱します。 </li><li>右の顕微鏡の手順の前に腹腔内ケタミンの注射（100mg / kg）およびキシラジン（5mg / kg）でマウスを麻酔。別の麻酔は、 例えば、2％イソフルラン吸入を用いることができます。反射刺激（しっかり圧でつま先やテールピンチ）に対する応答の喪失によって、適切な麻酔を確認してください。 </li><li>シェーバーを使用して腹部を脱毛するとエタノール70％で飽和したガーゼで抜け毛を取り除きます。 </li><li>麻酔下にある間、乾燥を防ぐために、マウスの目に獣医軟膏を適用します。 </li></ol> 2.手術<ol><li> 70％エタノールを用いて腹部を滅菌します。この方法は、滅菌方法ではなく、実験の終了時に致死的である可能性があります。 </li><li>正中開腹術を実行します。小さな、湾曲鉗子と小さなハサミを使用して腹部の皮膚を開きます。エピを特定gastrical血管や血管を保護するために、白線の領域で腹膜を開きます。 </li><li>湿った組織を維持するために腹腔内に予め加温した生理食塩水を数滴を適用します。 </li><li> 9,10前に説明したように眼窩50μlのローダミン6G（1mg / ml）を注入することにより、蛍光ラベルの白血球と血小板。 </li><li>直径10cmのペトリ皿にileumandの場所それのループを体外に出すし、37℃を適用することにより、湿った組織を維持することを確認してくださいは、生理食塩水（0.9％）、他のすべての分を予め加温しました。 </li></ol> 3.生体顕微鏡<ol><li>顕微鏡の下にマウスを置き、200の直径を有する腸間膜静脈を持って - ビューの中央には300μm。目に見える脂肪周囲と容器を選択してください。 </li><li>それによって内皮の刺激を避ける腸間膜血管を触れないでください。慎重回腸ループを処理します。 </li><li>血液細胞 - 内皮相互作用を可視化倒立または正立顕微鏡、顕微鏡ソフトウェアを使用して、カメラで。マウスあたり4つの異なる静脈の1分間の録音血液細胞 - 内皮相互作用。 </li><li>イメージング実験の完了後に頸椎脱臼によりマウスを安楽死させます。 </li></ol> 4.分析<ol><li>分析をオフラインで実行し、すべてのパラメータに対して盲目に。任意の適切なソフトウェアプログラムを用いて分析を行います。 </li><li>管腔内の血液細胞の流れに焦点を当て、高時間分解能のシネクリップで安定しており、interindividually同等の血流状態（最大フレームレート）を確認してください。 </li><li>ローリング白血球数を定量化します。そのための静脈を介して垂直線を描画し、1分でこのラインを交差するすべての白血球をカウントします。 </li><li>一つの白血球を安定して内皮上を転動しながら、50μmの距離を通過するのに必要な時間を測定することにより、圧延速度を決定。これを行うには、50ミクロンtの距離で2つの垂直線を描きます静脈hrough。 <ol><li>代表的な白血球が必要な時間（マイクロメートル/秒）を介して、50ミクロンを分割しother.Calculate行への白血球の速度を得るために必要な時間を測定します。 </li></ol></li><li> 0.04ミリメートル2のフィールドに白血球の接着を測定します。これを行うには、静脈に200μmの辺の長さの正方形を描画します。 <ol><li>この正方形の中に、30秒間、目に見える動きとして定義されてしっかりと付着性白血球を、数えます。 </li></ol></li><li>血小板 - 白血球の相互作用を定量化するために、1つの白血球に結合した血小板の数を数えます。 </li></ol>

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The authors have nothing to disclose.

本明細書中で、我々は、in vivoでのマウスの腸間膜静脈の生体顕微鏡の製造及び性能を記載します。この方法は、私たちに生体内で直接白血球-内皮細胞および血小板-内皮相互作用11を観察する機会を与えてくれます。 内皮を炎症に対する初期応答として活性化されますと、ローリング、接着および遊出12で得られた白血球と血小板と相互作用します...

このプロトコルの目的は、炎症状態の下で白血球 - 内皮細胞および白血球、血小板の相互作用を直接観察するために生きているマウスにおける腸間膜静脈の生体内顕微鏡検査の簡略化された手法を記述することです。 生体顕微鏡は、炎症性白血球動員および機能を理解するために些細なしかし重要ではありません炎症状態1,2、下のin vivoでの白血球-内皮相互作用を研究するために開発されました。私たちはこのプロトコルで説明した方法は、以前に公開された刊行物に基づいて開発されました。公開されたものに成長して血栓3に並行して、血小板の取り込みを可視化と同様に、以前の4、体外に腸間膜静脈を透過光顕微鏡で観察されます。例えばラット5やマウスの肝臓およびラット6の精巣挙筋としてイントラ不可欠な顕微鏡の他の様々なモデルがあります。腸間膜静脈の生体内顕微鏡法が開発され、いくつかのグループによって以前の研究に適用されています。非神経セロトニンの欠損しているマウス、およびその血小板セロトニン薬理学的に長期的により枯渇したマウスとの調査結果を比較して - / - 我々は、野生型マウスとトリプトファンhydroxylase1（TPH1）との間の白血球動員の違いを観察するためにこの技術を使用していますアプリケーション選択的セロトニン再取り込み阻害剤フルオキセチン7。また、急性フルオキセチン処置8後に白血球-内皮相互作用を検討しました。 このプロトコルでは、このモデルは迅速に行うことができるので、腸間膜静脈の生体内顕微鏡検査に焦点を当てて、白血球 - 内皮細胞および血小板 - 白血球の相互作用の有効な測定を可能にします。これは、はるかに困難な、時間のかかる、骨、肝臓、皮膚、または精巣挙筋のような他の臓器の生体内顕微鏡検査です。モードここで説明するlは、このようなリポ多糖の腹腔内注射のような炎症性刺激、とそれをchallengeing後の炎症性細胞間相互作用の再現性の評価に最適です。

<table><tbody><tr><td>Rhodamine 6G</td><td>Sigma-Aldrich</td><td>&amp;nbsp;989-38-8</td><td></td></tr><tr><td>Lipopolysaccharides from Escherichia coli 055:B5</td><td>Sigma-Aldrich</td><td>L2637&amp;nbsp;</td><td></td></tr></tbody></table>

intravital microscopy of leukocyte-endothelial and platelet-leukocyte interactions in mesenterial veins in mice

生体顕微鏡は、生きている動物における異なる領域と血管内の異なるプロセスを研究するために使用することができる方法です。このプロトコルでは、我々は腸間膜静脈の生体顕微鏡を説明します。これは、再現性のある結果がインビボで白血球-内皮相互作用を示すと共に、短時間で行うことができます。我々は、内皮細胞のLPSチャレンジ後の炎症の設定について説明します。しかし、このモデルでは1は、細菌の化学的または生物学的のように、炎症性状態の多くの異なる種類を適用し、薬剤の投与と生きている動物に対するその直接効果と白血球動員への影響を調査することができます。このプロトコルは、異なるマウスの処置および血管における炎症反応に及ぼす影響の数に正常に適用されました。ここで、我々は、蛍光ローダミン6Gでこれらを標識することにより、白血球と血小板の可視化について説明します。さらに、特定のイメージングは​​、PEとすることができます対象となる蛍光標識された分子を用いてrformed。

このプロトコールに記載された実験設定の概要を図1に示されている。これは、生体顕微鏡で観察することができる、体外回腸ループと血管を有するマウスを示す。 図2に起因して、未処理の動物における活性化の特定の程度を示します手順自体。しかし、ほとんどない低速圧延またはLPS処置マウスと比較して、白血球の強固な接着性がある。また、野生型マウスのい...

Watch this Scientific Journal Video about Intravital Microscopy of Leukocyte-endothelial and Platelet-leukocyte Interactions in Mesenterial Veins in Mice at JoVE.com

Intravital Microscopy of Leukocyte-endothelial and Platelet-leukocyte Interactions in Mesenterial Veins in Mice

This simplified application of intravital microscopy of mesentery veins in mice can be used in different models of inflammation to observe leukocyte-endothelial and platelet-leukocyte interactions.

マウスの腸間膜静脈内の生体顕微鏡白血球 - 内皮及び血小板 - 白血球の相互作用

Intravital microscopy is a method that can be used to investigate different processes in different regions and vessels in living animals. In this ...

intravital-microscopy-leukocyte-endothelial-platelet-leukocyte

Research

JoVE Journal

Immunology and Infection

12.1K ビュー.  Heart Center, University of Freiburg. This simplified application of intravital microscopy of mesentery veins in mice can be used in different models of inflammation to observe leukocyte-endothelial and platelet-leukocyte interactions.

ビデオ: Intravital Microscopy of Leukocyte-endothelial and Platelet-leukocyte Interactions in Mesenterial Veins in Mice

Intravital Microscopy of the Mouse Brain Microcirculation using a Closed Cranial Window

This experimental model was designed to assess the mouse pial microcirculation during acute and chronic, physiological and pathophysiological hemodynamic, inflammatory and metabolic conditions, using in vivo fluorescence microscopy. A closed cranial window is placed over the left parieto-occipital cortex of the mice. Local microcirculation is recorded in real time through the window using epi and fluorescence illumination, and measurements of vessels diameters and red blood cell (RBC) velocities are performed. RBC velocity is measured using real-time cross-correlation and/or fluorescent-labeled erythrocytes. Leukocyte and platelet adherence to pial vessels and assessment of perfusion and vascular leakage are made with the help of fluorescence-labeled markers such as Albumin-FITC and anti-CD45-TxR antibodies. Microcirculation can be repeatedly video-recorded over several days. We used for the first time the close window brain intravital microscopy to study the pial microcirculation to follow dynamic changes during the course of Plasmodium berghei ANKA infection in mice and show that expression of CM is associated with microcirculatory dysfunctions characterized by vasoconstriction, profound decrease in blood flow and eventually vascular collapse.

Intravital microscopy to follow temporal and spatial hemodynamic and inflammatory events in the pial microcirculation.

クローズ頭蓋窓を使用してマウス脳の微小循環の生体顕微鏡

This experimental model was designed to assess the mouse pial microcirculation during acute and chronic, physiological and pathophysiological hemodynamic, ...

神経科学

Intravital Microscopy of the Inguinal Lymph Node

Lymph nodes (LN's), located throughout the body, are an integral component of the immune system. They serve as a site for induction of adaptive immune response and therefore, the development of effector cells. As such, LNs are key to fighting invading pathogens and maintaining health. The choice of LN to study is dictated by accessibility and the desired model; the inguinal lymph node is well situated and easily supports studies of biologically relevant models of skin and genital mucosal infection.
The inguinal LN, like all LNs, has an extensive microvascular network supplying it with blood. In general, this microvascular network includes the main feed arteriole of the LN that subsequently branches and feeds high endothelial venules (HEVs). HEVs are specialized for facilitating the trafficking of immune cells into the LN during both homeostasis and infection. How HEVs regulate trafficking into the LN under both of these circumstances is an area of intense exploration. The LN feed arteriole, has direct upstream influence on the HEVs and is the main supply of nutrients and cell rich blood into the LN. Furthermore, changes in the feed arteriole are implicated in facilitating induction of adaptive immune response. The LN microvasculature has obvious importance in maintaining an optimal blood supply to the LN and regulating immune cell influx into the LN, which are crucial elements in proper LN function and subsequently immune response. 
The ability to study the LN microvasculature in vivo is key to elucidating how the immune system and the microvasculature interact and influence one another within the LN. Here, we present a method for in vivo imaging of the inguinal lymph node. We focus on imaging of the microvasculature of the LN, paying particular attention to methods that ensure the study of healthy vessels, the ability to maintain imaging of viable vessels over a number of hours, and quantification of vessel magnitude. Methods for perfusion of the microvasculature with vasoactive drugs as well as the potential to trace and quantify cellular traffic are also presented. 
Intravital microscopy of the inguinal LN allows direct evaluation of microvascular functionality and real-time interface of the direct interface between immune cells, the LN, and the microcirculation. This technique potential to be combined with many immunological techniques and fluorescent cell labelling as well as manipulated to study vasculature of other LNs.

A technique for performing intravital microscopy of the inguinal lymph node (LN) is outlined. Such technique allows for real-time, in vivo study of the lymph node microvasculature and structure both during homeostasis and infection. This technique can be adapted to cell trafficking studies and to other lymph node sites.

鼠径リンパ節の生体顕微鏡

Lymph nodes (LN's), located throughout the body, are an integral component of the immune system. They serve as a site for induction of adaptive immune ...

免疫・感染学

Real-time Imaging of Heterotypic Platelet-neutrophil Interactions on the Activated Endothelium During Vascular Inflammation and Thrombus Formation in Live Mice

Interaction of activated platelets and leukocytes (mainly neutrophils) on the activated endothelium mediates thrombosis and vascular inflammation.1,2 During thrombus formation at the site of arteriolar injury, platelets adherent to the activated endothelium and subendothelial matrix proteins support neutrophil rolling and adhesion.3 Conversely, under venular inflammatory conditions, neutrophils adherent to the activated endothelium can support adhesion and accumulation of circulating platelets. Heterotypic platelet-neutrophil aggregation requires sequential processes by the specific receptor-counter receptor interactions between cells.4 It is known that activated endothelial cells release adhesion molecules such as von Willebrand factor, thereby initiating platelet adhesion and accumulation under high shear conditions.5 Also, activated endothelial cells support neutrophil rolling and adhesion by expressing selectins and intercellular adhesion molecule-1 (ICAM-1), respectively, under low shear conditions.4 Platelet P-selectin interacts with neutrophils through P-selectin glycoprotein ligand-1 (PSGL-1), thereby inducing activation of neutrophil &beta;2 integrins and firm adhesion between two cell types. Despite the advances in in vitro experiments in which heterotypic platelet-neutrophil interactions are determined in whole blood or isolated cells,6,7 those studies cannot manipulate oxidant stress conditions during vascular disease. In this report, using fluorescently-labeled, specific antibodies against a mouse platelet and neutrophil marker, we describe a detailed intravital microscopic protocol to monitor heterotypic interactions of platelets and neutrophils on the activated endothelium during TNF-&alpha;-induced inflammation or following laser-induced injury in cremaster muscle microvessels of live mice.

Here we report an experimental technique of fluorescence intravital microscopy to visualize heterotypic platelet-neutrophil interactions on the activated endothelium during vascular inflammation and thrombus formation in live mice. This microscopic technology will be valuable to study the molecular mechanism of vascular disease and to test pharmacologic agents under pathophysiological conditions.

生きたマウスにおける血管炎症及び血栓形成の間に活性化内皮上に異型血小板、好中球の相互作用のリアルタイムイメージング

Interaction of activated platelets and leukocytes (mainly neutrophils) on the activated endothelium mediates thrombosis and vascular inflammation.1,2 ...

Imaging Neutrophils and Monocytes in Mesenteric Veins by Intravital Microscopy on Anaesthetized Mice in Real Time

Efficient immune response is dependent on rapid mobilization of blood leukocytes to the site of infection or injury. Investigating leukocyte migration in vivo is crucial for understanding the molecular basis of leukocyte transendothelial migration and interaction with vascular endothelium. One powerful approach involves intravital microscopy on transgenic mice expressing fluorescent proteins in cells of interest.
Here we present a protocol for imaging monocytes and neutrophils in the CX3CR1gfp/wt mouse i.v. injected with orange dye-labeled neutrophils with an inverted confocal microscope. Time-lapse movies gathered from 30 min&#160;to several hours of imaging allow the analysis of leukocyte behavior in mesenteric veins under both steady state and inflammatory conditions. We also describe the steps to locally induce blood vessel inflammation with TLR2/TLR1 agonist Pam3SK4 and monitor the subsequent recruitment of neutrophils and monocytes.
The presented technique can also be used to monitor other populations of leukocytes and investigate molecules implicated in leukocyte recruitment or trafficking using other stimuli or transgenic mice.

We detail a protocol to monitor the behavior of neutrophils and monocytes in mesenteric veins under steady state and inflammatory conditions using intravital confocal microscopy on anaesthetized mice.

リアルタイムで麻酔したマウスに生体顕微鏡により腸間膜静脈に好中球および単球のイメージング

Efficient immune response is dependent on rapid mobilization of blood leukocytes to the site of infection or injury. Investigating leukocyte migration in ...

Intravital Microscopy of Monocyte Homing and Tumor-Related Angiogenesis in a Murine Model of Peripheral Arterial Disease

The therapeutic goal for peripheral arterial disease and ischemic heart disease is to increase blood flow to ischemic areas caused by hemodynamic stenosis. Vascular surgery is a viable option in selected cases, but for patients without indications for surgery such as progression to rest pain, critical limb ischemia, or major disruptions to life or work, there are few possibilities for mitigating their disease. Cell therapy via monocyte-enhanced perfusion through the stimulation of collateral formation is one of a few non-invasive options.
Our group examines arteriogenesis after monocyte transplantation into mice using the hindlimb ischemia model. Previously, we have demonstrated improvement in hindlimb perfusion using tetanus-stimulated syngeneic monocyte transplantation. In addition to the effects on the collateral formation, tumor growth could be affected by this therapy as well. To investigate these effects, we use a basement membrane-like matrix mouse model by injecting the extracellular matrix of the Engelbreth-Holm-Swarm sarcoma into the flank of the mouse, after occlusion of the femoral artery.
After the artificial tumor studies, we use intravital microscopy to study in vivo tumor-angiogenesis and monocyte homing within collateral arteries. Previous studies have described the histological examination of animal models, which presupposes subsequent analysis to post-mortem artifacts. Our approach visualizes monocyte homing to areas of collateralization in real time sequences, is easy to perform, and investigates the process of arteriogenesis and tumor angiogenesis in vivo.

Monocytes are important mediators of arteriogenesis in the context of peripheral arterial disease. Using a basement membrane-like matrix and intravital microscopy, this protocol investigates monocyte homing and tumor-related angiogenesis after monocyte injection in the femoral artery ligation murine model.

単球のホーミングと末梢動脈疾患のマウスモデルにおける腫瘍関連血管新生の生体顕微鏡

The therapeutic goal for peripheral arterial disease and ischemic heart disease is to increase blood flow to ischemic areas caused by hemodynamic ...

医学

Leukocyte Infiltration of Cremaster Muscle in Mice Assessed by Intravital Microscopy

Intravital microscopy (IVM) is widely used to monitor physiological and pathophysiological processes within the leukocyte recruitment cascade in vivo. The current protocol represents a practical and reproducible method to visualize the leukocyte endothelium interaction leading to leukocyte recruitment in skeletal muscle derived tissue within the intact organism of the mouse. The model is applicable to all fields of research that focus on granulocyte activation and their role in disease.
We provide a step by step protocol to guide through the method and to highlight potential pitfalls and technical difficulties. The protocol covers the following aspects: experimental settings and required material, anesthesia of the mouse, dissection of the cremaster muscle as well as tracheal and carotid cannulation, IVM recordings and offline analysis. Data formats like adherent leukocytes, rolling flux (RF) and rolling flux fraction (RFF) are explained in detail and appropriate applications are discussed. Representative results from dystrophin deficient mdx mice are provided in the results section.
IVM is a powerful tool to assess leukocyte recruitment in an in vivo setting; however, delineating for example endothelial and leukocyte function may require a combination with ex vivo setups like flow chamber experiments. Furthermore, the genetic background of animals of interest may greatly influence baseline recruitment, requiring individual fine tuning of the protocol provided. Despite its limitations, IVM may serve as a platform to readily translate in vitro findings into a living vertebrate organism.

Here, we show how to perform intravital microscopy on post-capillary venules of the mouse cremaster muscle. Commonly applied to different models of inflammation and sepsis, particularly those induced by chemokines and cytokines, we highlight its relevance in the study of muscolopathies involving exaggerated muscular leukocyte infiltration.

インビタル顕微鏡で評価されたマウスにおけるクレマスター筋の白血球浸潤

Intravital microscopy (IVM) is widely used to monitor physiological and pathophysiological processes within the leukocyte recruitment cascade in vivo. The ...

Multiphoton Intravital Imaging for Monitoring Leukocyte Recruitment during Arteriogenesis in a Murine Hindlimb Model

Arteriogenesis strongly depends on leukocyte and platelet recruitment to the perivascular space of growing collateral vessels. The standard approach for analyzing collateral arteries and leukocytes in arteriogenesis is ex vivo (immuno-) histological methodology. However, this technique does not allow the measurement of dynamic processes such as blood flow, shear stress, cell-cell interactions, and particle velocity. This paper presents a protocol to monitor in vivo processes in growing collateral arteries during arteriogenesis utilizing intravital imaging. The method described here is a reliable tool for dynamics measurement and offers a high-contrast analysis with minimal photo-cytotoxicity, provided by multiphoton excitation microscopy. Prior to analyzing growing collateral arteries, arteriogenesis was induced in the adductor muscle of mice by unilateral ligation of the femoral artery.
After the ligation, the preexisting collateral arteries started to grow due to increased shear stress. Twenty-four hours after surgery, the skin and subcutaneous fat above the collateral arteries were removed, constructing a pocket for further analyses. To visualize blood flow and immune cells during in vivo imaging, CD41-fluorescein isothiocyanate (FITC) (platelets) and CD45-phycoerythrin (PE) (leukocytes) antibodies were injected intravenously (i.v.) via a catheter placed in the tail vein of a mouse. This article introduces intravital multiphoton imaging as an alternative or in vivo complementation to the commonly used static ex vivo (immuno-) histological analyses to study processes relevant for arteriogenesis. In summary, this paper describes a novel and dynamic in vivo method to investigate immune cell trafficking, blood flow, and shear stress in a hindlimb model of arteriogenesis, which enhances evaluation possibilities notably.

The recruitment of leukocytes and platelets constitutes an essential component necessary for the effective growth of collateral arteries during arteriogenesis. Multiphoton microscopy is an efficient tool for tracking cell dynamics with high spatio-temporal resolution in vivo and less photo-toxicity to study leukocyte recruitment and extravasation during arteriogenesis.

マウス後肢モデルにおける動脈形成中の白血球動員モニタリングのための多光子生体内イメージング

Arteriogenesis strongly depends on leukocyte and platelet recruitment to the perivascular space of growing collateral vessels. The standard approach for ...

<meta charset="utf8"></head><body>I/R損傷時の白血球接着とNET形成における内皮PAR1の役割

Visualization of Neutrophil Extracellular Traps in Mesenteric Venules After Mesenteric Ischemia-Reperfusion Injury via Intravital Microscopy

Intestinal ischemia-reperfusion (I/R) injury is an acute condition characterized by tissue damage resulting from restricted blood flow to the mesenteric vessels, leading to both local and systemic pathologies with a poor prognosis. Both ischemia and reperfusion trigger a series of cellular and molecular responses, with inflammatory cells serving as key regulators of the pathology. These interactions with the ischemic endothelium are mediated by multiple adhesion receptors. Several animal models have been established to mimic this pathology and investigate the involved molecular pathways. In this study, a microsurgical model of I/R injury is combined with intravital microscopy to visualize leukocyte rolling, adhesion, and neutrophil extracellular trap (NET) formation. This model is applied to transgenic mice deficient in endothelial PAR1 (F2r) to assess the impact of PAR1 on leukocyte rolling and NET formation 1 h after ischemia and immediately following reperfusion. In vivo, Acridine Orange leukocyte staining was employed, and NETs were visualized using a nucleic acid stain. Interestingly, reduced leukocyte adhesion and NET formation were observed in mice lacking the endothelial PAR1 receptor. This model enables the in vivo analysis of key regulators involved in I/R injury.

<meta charset="utf8"></head><body>腸間膜虚血再灌流障害後の腸間小静脈における好中球細胞外トラップの生体内顕微鏡による可視化

This protocol presents a method for inducing mesenteric ischemia-reperfusion injury and visualizing neutrophil extracellular traps (NETs) in mesenteric venules using intravital microscopy. Blood flow in the superior mesenteric artery was restricted for 1 h, followed by reperfusion, allowing direct quantification of leukocyte adhesion and NETosis.

腸間膜虚血再灌流障害後の腸間小静脈における好中球細胞外トラップの生体内顕微鏡による可視化

Intestinal ischemia-reperfusion (I/R) injury is an acute condition characterized by tissue damage resulting from restricted blood flow to the mesenteric ...

マウスの腸間膜静脈内の生体顕微鏡白血球 - 内皮及び血小板 - 白血球の相互作用

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クローズ頭蓋窓を使用してマウス脳の微小循環の生体顕微鏡

鼠径リンパ節の生体顕微鏡

生きたマウスにおける血管炎症及び血栓形成の間に活性化内皮上に異型血小板、好中球の相互作用のリアルタイムイメージング

リアルタイムで麻酔したマウスに生体顕微鏡により腸間膜静脈に好中球および単球のイメージング

単球のホーミングと末梢動脈疾患のマウスモデルにおける腫瘍関連血管新生の生体顕微鏡

インビタル顕微鏡で評価されたマウスにおけるクレマスター筋の白血球浸潤

マウス後肢モデルにおける動脈形成中の白血球動員モニタリングのための多光子生体内イメージング

腸間膜虚血再灌流障害後の腸間小静脈における好中球細胞外トラップの生体内顕微鏡による可視化

腸間膜虚血再灌流障害後の腸間小静脈における好中球細胞外トラップの生体内顕微鏡による可視化

腸間膜虚血再灌流障害後の腸間小静脈における好中球細胞外トラップの生体内顕微鏡による可視化