Name: in vivo imaging of cx3cr1gfp/gfp reporter mice with spectral-domain optical coherence tomography and scanning laser ophthalmoscopy
Uploaded: 2017-11-11T14:00:00
Description: Watch this Scientific Journal Video about In Vivo Imaging of Cx3cr1gfp/gfp Reporter Mice with Spectral-domain Optical Coherence Tomography and Scanning Laser Ophthalmoscopy at JoVE.com

この作品は、スイスの全米科学財団 (SNSF; #320030_156019) の補助金によって支えられました。著者らは、ハイデルベルク工学 GmBH は、ドイツから金融サポートを受け取った。

すべての手順で BALB/c 雌雄成体 Cx3cr1 のプロモーター gfp を表現するいた使用 24。マウスは、眼科と視覚に関する研究における動物の使用の ARVO 声明に従って扱われ、すべてのプロシージャは、動物福祉に関するスイス連邦政府の規則によるとスイス政府から承認されました。マウス塩酸メデトミジン (0.75 mg/kg) とケタミン (45 の mg/kg の皮下注射で麻酔をかけられまし...

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本稿は、網膜 B-スキャンの買収と同じ撮像セッションでマウス網膜における gfp 陽性ミクログリア分布のイメージングのためのプロトコルを示します。SD 10 月と SLO はますます時間10,14,17,18,21に網膜の変化の情報を提供するために網膜疾患の動物モデルで使用されます。?...

実験眼科で網膜の病理検査では組織学的手法を使用して評価、通常。しかし、組織は動物 euthanization を必要とし、組織の実際のプロパティに変更を引き起こす可能性があります。SD 10 月と SLO が定期的に眼科で使用される臨床診断用といくつかの網膜疾患網膜色素変性症11 や前部虚血性視神経症10、糖尿病黄斑浮腫9などの監視のため.SD 10 月 SLO は、非侵襲的手法、さら、介入を使わないで瞳孔を通して可視化、網膜の高解像度画像を生成します。SD-10 月は、SLO 蛍光網膜の立体の高コントラストの画像を生成するデータを収集中に、網膜の断面像を作成する後方散乱データを集めることによって網膜の厚さと網膜の構造の情報を提供します。今日では、両方の技術はますます小さい齧歯動物12,13,14,15 (または) を使用もゼブラフィッシュ16,17実験眼科で使用され、することができます。両方質的・量的情報12,17,18,19,20,21を提供します。
Lipofuscins のような内因性蛍光物質の蓄積や乳頭部のドルーゼン、網膜の形成が自動蛍光信号として SLO で視覚化できます。この機能は、診断と網膜症加齢黄斑変性症や網膜色素変性症22,23などの監視のための貴重な技術を SLO になります。実験眼科でレポーター マウスの行の特定のセル型の検出の自動蛍光 (AF) を使用できます。たとえば、 Cx3cr124のプロモーターの gfp 発現のヘテロ接合体マウスは通常の網膜におけるミクログリア細胞の生体内可視化およびミクログリア/マクロファージの調査のために有利です網膜疾患21のダイナミクス。ミクログリアは、網膜のマクロファージ組織恒常性と損傷1,25,26の時に組織の修復に重要な役割を果たしています。網膜におけるミクログリアの活性化が、網膜の損傷、虚血、変性、網膜疾患2,3,4,5,におけるこれらの細胞の役割を示唆している報告されています。6。
この議定書の目的Cx3cr1gfp/gfpマウス網膜を使って gfp 陽性ミクログリア細胞の可視化、網膜イメージングおよび SD-OCT による網膜の厚さの測定のための比較的簡単なメソッドを記述することです。SLO (ハイデルベルク Spectralis HRA + OCT システム)。このプロトコルはマウスの各種の行の健康または病気の網膜のイメージングや膜厚計測の利用できます。さらに、同定及び定量化のミクログリア数字と SLO21を使用して網膜におけるミクログリアの活性化の形態計測学的解析を実行できます。ミクログリア細胞は網膜27,28,29を含む中枢神経系 (CNS)、変性疾患に関連付けられます。したがって、この議定書で使用される 2 つの方法を組み合わせることによってミクログリア分布と網膜変性の相関を行うことができますは、重症度監視を容易にすることができますをまたは生体内で治療の有効性に近づきます。

<table>
	<tbody>
		<tr>
			<td>Spectralis Imaging system (HRA+OCT)</td>
			<td>Heidelberg Engineering, Germany</td>
			<td>N/A</td>
			<td>ophthalmic imaging platform system</td>
		</tr>
		<tr>
			<td>Heidelberg Eye Explorer</td>
			<td>Heidelberg Engineering, Germany</td>
			<td>N/A</td>
			<td>Version 1.9.13.0</td>
		</tr>
		<tr>
			<td>78D standard ophthalmic non-contact slit lamp lens</td>
			<td>Volk Optical Inc., Ohio, USA</td>
			<td>V78C</td>
			<td></td>
		</tr>
		<tr>
			<td>Spectralis wide angle 55&#176; lens</td>
			<td>Heidelberg Engineering, Germany</td>
			<td>50897-002</td>
			<td></td>
		</tr>
		<tr>
			<td>ultra widefield 102&#176; lens</td>
			<td>Heidelberg Engineering, Germany</td>
			<td>50117-001</td>
			<td></td>
		</tr>
		<tr>
			<td>medetomidine hydrochloride 1 mg/mL (Domitor)</td>
			<td>Provet AG, Lyssach, Switzerland</td>
			<td>Swissmedic Nr. 50&#39;590 - ATCvet: QN05CM91</td>
			<td>anesthetic/analgesic</td>
		</tr>
		<tr>
			<td>ketamine 50mg/ml (Ketalar)</td>
			<td>Parke-Davis, Zurich, Switzerland</td>
			<td>72276388</td>
			<td>anesthetic</td>
		</tr>
		<tr>
			<td>tropicamide 0.5% + phenylephrine HCl 2.5% (Augentropfen mix)</td>
			<td>ISPI, Bern, Switzerland</td>
			<td>N/A</td>
			<td>pupil dilation</td>
		</tr>
		<tr>
			<td>Omnican Insulin-50 0.5 ml G30 0.3 x 12mm</td>
			<td>B. Braun Mesungen AG, Carl-Braun-Stra&#223;e, Germany</td>
			<td>9151125</td>
			<td></td>
		</tr>
		<tr>
			<td>hydroxypropylmethylcellulose (Methocel 2%)</td>
			<td>OmniVision, Neuhausen, Switzerland</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>+4 dpt rigid gas permeable contact lens</td>
			<td>Quantum I, Bausch + Lomb Inc., Rochester, NY</td>
			<td>N/A</td>
			<td>Base Curve: 7.20 to 8.40 mm 
			Diameter: 9.00 / 9.60 / 10.20 mm 
			Power: -25.00 to +25.00 Diopters</td>
		</tr>
		<tr>
			<td>balanced salt solution (BSS)</td>
			<td>Inselspital, Bern, Switzerland</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>silicon forceps</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>atipamezole 5 mg/mL (Antisedan)</td>
			<td>Provet AG, Lyssach, Switzerland</td>
			<td>N/A</td>
			<td>&#945;2 adrenergic receptor antagonist</td>
		</tr>
		<tr>
			<td>GraphPad Prism 7</td>
			<td>GraphPad Software, Inc, San Diego, CA, USA</td>
			<td>N/A</td>
			<td>statistical analysis software</td>
		</tr>
	</tbody>
</table>

in vivo imaging of cx3cr1gfp/gfp reporter mice with spectral-domain optical coherence tomography and scanning laser ophthalmoscopy

スペクトル領域光干渉断層法 (SD OCT) と走査レーザー眼底検査 (SLO) は、実験眼科で幅広く使用されています。現在のプロトコルを発現するマウス緑蛍光蛋白質 (gfp) Cx3cr1のプロモーター (BALB/c-Cx3cr1gfp/gfp) ミクログリア細胞体内の網膜のイメージのために使用されました。ミクログリアは網膜のマクロファージであり、いくつかの網膜疾患1,2,3,4,5,6に関与しています。このプロトコルは、網膜 B-スキャンの SD-10 月、SLO は生体内で、 Cx3cr1gfp/gfpマウスのミクログリア細胞分布のイメージングと眼画像処理プラットフォーム システムを使用して生成の詳細なアプローチを提供します。プロトコルは、レポーター マウスのいくつかの行で使用することができます。ただし、ここで示されるプロトコルをいくつかの制限があります。まず、両方の SLO と SD 10 月では、高分解能モードで使用する場合高軸方向の解像度が横方向の解像度を持つデータを収集が低い (3.5 μ m と 6 μ m、それぞれ)。さらに、SLO のフォーカスと彩度レベルはパラメーターの選択や目の適切な配置に依存です。さらに、マウスの人間の患者はマウス眼の高い合計光学力のために挑戦のために設計されたデバイスを使用してください。 人間の目と比較してください。これはとりわけマウス レンズによって倍率に依存でもある横倍率誤り7、可能性があります。しかし、軸のスキャン位置にもかかわらず、横倍率に依存して、軸の SD 10 月測定は正確な8。

ここで提示されたプロトコルを使用して、SD OCT スキャンし、SLO 画像は、同じ撮像セッションでCx3cr1gfp/gfpマウスから得られました。図 3には、代表者 SD 10 月 gfp 陽性ミクログリア細胞が視覚化される 55 ° または 102 ° レンズで得られる 30 ° または 55 ° レンズ (図 3 a) と代表的な SLO 画像で得られた単一のスキ...

Watch this Scientific Journal Video about In Vivo Imaging of Cx3cr1gfp/gfp Reporter Mice with Spectral-domain Optical Coherence Tomography and Scanning Laser Ophthalmoscopy at JoVE.com

In Vivo Imaging of Cx3cr1gfp/gfp Reporter Mice with Spectral-domain Optical Coherence Tomography and Scanning Laser Ophthalmoscopy

このプロトコルを記述する方法高解像度イメージング スペクトル領域光干渉断層法などと小さい齧歯動物, 眼科画像処理プラットフォーム システムを使用してに関する情報を取得するに活用できるレーザー眼底をスキャン網膜厚とミクログリア細胞の分布、それぞれ。

生体内でスペクトル領域光干渉断層法と走査レーザー眼底Cx3cr1gfp/gfpレポーター マウスの画像

Spectral domain optical coherence tomography (SD-OCT) and scanning laser ophthalmoscopy (SLO) are extensively used in experimental ophthalmology. In the ...

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In Vivo Imaging of Cx3cr1gfp/gfp Reporter Mice with Spectral-domain Optical Coherence Tomography and Scanning Laser Ophthalmoscopy

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We describe a multi-angle rotational optical imaging (MAROI) system for in vivo monitoring of physiopathological processes labeled with a fluorescent ...

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Stroke is the third leading cause of death among Americans 65 years of age or older1. The quality of life for patients who suffer from a stroke fails to ...

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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

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ナノボディ及び抗体ベースの検証<em&gt;インビボ</em使用してマウスにおける&gt;腫瘍異種移植NIRFイメージング実験<em&gt;エクスビボ</em&gt;フローサイトメトリーおよび顕微鏡

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生体内でスペクトル領域光干渉断層法と走査レーザー眼底Cx3cr1gfp/gfpレポーター マウスの画像

生体内でスペクトル領域光干渉断層法と走査レーザー眼底Cx3cr1^gfp/gfpレポーターマウスの画像