この作品は、国立眼研究所 4K12EY022299 の寛大なサポートによって支えられた (ワイエムピー)、戦いの視力国際網膜研究財団 FFS GIA16002 (ワイエムピー)、失明を防ぐための研究から無制限の部門レベルのサポートとミシガン大学眼科学および視覚科学。この作品を利用するコア センターで、ビジョン研究国立眼研究所から P30 EY007003 によって資金を供給します。

ウサギは、アメリカ合衆国農務省 (USDA) に種が覆われています。生物医学研究での使用は厳しい規制に従う必要があります。すべてウサギ実験を行なった ARVO (視覚と眼科学研究協会) ステートメントに従って眼科と視覚に関する研究における動物の使用のため、大学による研究所動物プロトコルの承認後委員会委員の使用、ミシガン大学 (プロトコル PRO00006486、PI ヤニス Paulus) 動物 (UCUCA)。
1. システムの構成
<ol><li>光音響顕微鏡 (PAM)<ol>
<li>光パラメトリック発振 (OPO) レーザーの使用は PAM. 選択の適切な技術仕様の光源としてダイオード励起固体レーザー励起パルス繰り返しレート 1 kHz、パルス期間 3-6 ns、および可変波長 405-2600 nm のよう。</li>
		<li>570 でレーザーから発せられるビームを反映 (M1、M2)、2 つのミラーによって nm、電動回転ステージ、そして最後にフォーカス、フィルターにマウントされている半波長板の減衰器を通過し、ビーム コリメータ (図 1) によるコリメートします。ビーム コリメータのデザインを最適化します。ビーム コリメータの構成の例を含む焦点レンズ L1 (焦点距離 250 mm)、(直径 50 μ m)、ピンホールとコリメート レンズ L2 (焦点距離 30 mm)。</li>
		<li>平行光ビームスプリッター (BS1) 90/10 (反射/透過) の分割比で除算します。パルス-パルス レーザー エネルギーを監視するためフォト ダイオード、透過部分を記録します。ミラー (M3) とダイクロイック ミラー (DM) とラスター スキャンによる反射部分をそらす二次元検流計を使用して連続して。ガルバノはスペクトル ドメイン (SD) と共有コンポーネント-OCT システム (後述)。</li>
		<li>配信望遠鏡でスキャンしたビームから成るスキャン レンズ (焦点距離 36 mm) と眼のレンズ (OL、焦点距離 10 mm) 最後にウサギ目光学系による眼底に集中し、。</li>
		<li>適切な技術仕様、例えば、中心周波数と針状の超音波トランスデューサーを選択 27 MHz、双方向 − 6 dB 帯域幅の 60%。励起光音響信号をキャプチャする中央の視軸を結膜との接触に配置します。</li>
		<li>超音波増幅器を用いた信号を増幅する (たとえば、57 dB 利得) 低域通過フィルターによってフィルター処理 (たとえば、カットオフ周波数 32 MHz)、200 MS/秒のサンプリング レートでの高速デジタイザーがデジタル化して。</li>
		<li>場所上記電源メーター、ウサギ目し、ANSI 安全に保つことは、角膜にレーザー パルス エネルギーを測定制限 160 570 nm38でニュージャージー州。Matlab から同期エレクトロニクスで制御レーザー シャッターを使用してレーザーの露出オーバーを避けるためにビームをブロックします。</li>
		<li>デジタイザーのデータ集録 (DAQ) ボードを介して、検流計、レーザーを同期します。プログラムは Matlab でシステム制御およびデータ取得用のソフトウェア。</li>
	</ol>
</li>
	<li>スペクトル領域光干渉断層法 (SD-10 月)<ol>
<li>参照の腕 (図 1)、眼科 (OL) スキャン レンズ (SL) の後、一枚のレンズ分散補償ガラス (DCG) を追加して市販システムに基づく SD OCT システムを適応します。変更により、OCT システムがウサギの眼の後部セグメントをイメージすることができます。</li>
		<li>ズーム住宅管を使ってサンプル腕の光路長との一致を確保するための参照の腕の長さを調整します。アイリスを使用して、最大の画像のコントラストを達成するためにウサギ眼底から後方散乱光強度での一致を保証するためのレトロな反映参照光の強度を制御します。</li>
		<li>発光ダイオード (LED) 外部光源として照明光とウサギ眼底のリアルタイム可視化のためのスキャン ヘッドにカプセル化電荷結合素子 (CCD) カメラを採用してください。</li>
	</ol>
</li>
</ol>2. システムの配置
<ol><li>ガルバノの位置の初期化、ファイバー コリメータ マウント、ビームスプリッター キューブのネジを調整することによって 10 月システムを合わせます。 注: 手順市販の OCT システムのマニュアルで利用でき、ここでは説明しません。この手順は主にファイバー コリメータ、参照アーム、OCT システムのパフォーマンスを最大化するスキャン レンズの正しい配置を確保するためです。</li>
	<li>空間的レーザー光をフィルターで除外し、最大限にレーザー エネルギーを送信または集光レンズの焦点の周りピンホールのx y、およびzの位置を調整します。高さ測定ツールを使用して、彼らが同じであることを確認するピンホールの前後には、レーザ光の高さを確認してください。</li>
	<li>チルト、偏心とコリメート レンズ フィルター処理されたビームをコリメートする L2 のz位置を調整します。ビームがおよそ同じであることを確認サイズと近傍界と遠方界で観測した時の高さ。</li>
	<li>Co 軸ミラーと DM の傾きを調整して PAM レーザービームと 10 月の光ビームを組み合わせます。この手順の後、PAM レーザーと 10 月の光が完全に一致する必要があり、ウサギ眼底をスキャン領域が同じ。</li>
	<li>角度調整と光パスに合わせて正しく OL レンズの偏心。一度完了したら、デュアル モダリティ システムのイメージングの準備が。 注: 自動視準メソッドをなど、これを達成するために使用し、入射光として同じ道を戻ることを確認するのに OL レンズ表面で反射する光を確認できます 1。</li>
</ol>3. ウサギの準備
<ol><li>動物実験施設と体の重量と動物の数など、個人情報を記録からニュージーランド白ウサギを取る。</li>
	<li>ボディ サポートおよび光学イメージング システム表にヘッド サポートを含むウサギ プラットフォームをマウントします。ボディ サポートに水を循環させ暖房毛布を置くし、循環水の温度を実験的回復期間のウサギの体温を暖かい保つために 38 ° C に設定します。</li>
	<li>前のプロシージャきゅうしょを記録全体的に動物の状態、粘膜色、心拍数、呼吸数、および直腸体温を含む。ケタミン (40 mg/kg) とキシラジン投与 (IM) 経由 (5 mg/kg) の混合物とウサギの麻酔し、ケタミン (スケジュール III 薬物) の使用率を記録します。心拍数、呼吸数、および全体的な状態を確認することによって麻酔レベルを確認します。</li>
	<li>トロピカミド 1% 眼とフェニレフリン塩酸塩 2.5% 眼の一滴を使用してウサギ瞳孔を広げます。</li>
	<li>鏡を使用して、まぶたを保持する方法のうち、角膜を湿らせて目の潤滑剤の滴を適用します。局所テトラカインのドロップを植え付けるイメージ作成手順の前に目の 0.5%。 注: 長いプロシージャまたはプロシージャ動物に可能な不快感を与えるウサギ動物の快適さを確保するための実験の前にメロキシカムの皮下注射。</li>
</ol>4. SD OCT イメージング
<ol><li>ウサギの臨床眼底カメラのイメージング プラットフォーム転送し、イメージング セッション 10 月前に 50 度の眼底、無料、フリー、赤、蛍光画像を取る。こう目の光透過性をチェックし、眼底血管形態と髄質光線網膜血管、視神経などのランドマークを認識できます。</li>
	<li>ウサギの OCT システムのプラットフォームに転送し、約 OL の下の目の 1 つの位置にその姿勢を調整します。LED ライトを使用して目を照らします。 注: 技術の臨床の翻訳を容易にするウサギ目は安定、他のメソッドを使用して、ちょうど任意固定せずヘッド サポートにウサギの頭を置きます。</li>
	<li>10 月ソフトウェアを開き、まず眼底の CCD カメラの画像を確認します。カメラの視野 (FOV) 内にある脈絡膜血管や網膜血管が興味 (ROIs) の領域を確保するため必要に応じてヘッド サポートの角度と高さを細かく調整します。 注: ウサギが良い麻酔レベル下にある場合は、1 つの連続撮像セッションできますウサギの頭を再調整を必要とせず 10 分限りに最後。</li>
	<li>興味の 10 月の B-スキャンを表し、スキャンを開始する直線を描画します。OCT 画像を視覚化し、鮮明な画像を取得する 10 月ソフトウェアの分散補償係数を最適化する参照のアームの長さを調整します。 注: 参照のアームの長さを調整するとき、2 つのミラー化された 10 月画像 1 つ他の後で表示されます。適切なイメージは、眼底の解剖学の知識に基づいて識別できた。</li>
	<li>ピクセルの数と、平均値などのデータ集録パラメーターを設定し、画像を保存します。</li>
	<li>呼吸と麻酔レベルと動物を推定するウサギの心拍数を観察の快適さ。長いセッションの補足ケタミン又はイソフルラン吸入の 3 分の 1 の線量と見なされる可能性気管内挿管、V-ゲル、または顔マスク麻酔の必要な場合の平面を維持するために。</li>
	<li>角膜表面の脱水と角膜表層点状表層上皮性角膜症を防ぐために実験中に洗眼 2 分ごとに角膜をすすいでください。監視および記録の動物きゅうしょ 15 分毎。</li>
</ol>5. PAM イメージング
<ol><li>適切なレーザー安全ゴーグルを着用、OPO レーザーを入れます。</li>
	<li>PAM 制御ソフトウェアを起動、ターゲット発色団を (例えば、 570 nm ヘモグロビン) の吸収ピークの 1 つにレーザーの波長を調整、検流計、ようにウサギ角膜前にレーザー エネルギーをモニターの位置の初期化ANSI 安全基準以下です。</li>
	<li>三次元 (3 D) 翻訳段階に超音波探触子をマウントし、眼底を指しているウサギの結膜に接触探触子の先端位置します。良いカップル探触子の先端とウサギの結膜に目潤滑油をドロップを使用します。</li>
	<li>LED イルミネーション ライトをオンにし、Matlab ソフトウェアを介してウサギ眼底を可視化します。</li>
	<li>セット (網膜血管や脈絡膜血管) スキャンの ROI は、センター、物理サイズを含みます。レーザー シャッターを開き、ビームの B-スキャンを開始します。約探触子を配置は、1 つは検出光音響を見ることができるはず、オシロ スコープに信号。いない場合は、角膜の別の領域をスキャンまたは他の目に切り替え、上記のプロセスを繰り返すに目の位置をわずかに調整します。</li>
	<li>検出光音響信号をオシロ スコープと B スキャン全体に沿って信号強度を最大化する探触子の位置を微調整を観察します。 メモ: ため限られたビーム幅、超音波探触子通常が小さな FOV41。この手順では、PAM の最終のイメージの背景の変調を決定します。ミスアライメントが PAM 異種の背景を持つイメージにつながるし、画質を大きく劣化させます。</li>
	<li>データ集録パラメーターを設定します。これはピクセルの数が含まれています (e.g。、256 × 256 ピクセル)、サンプリング レート (e.g。、200 MS/秒)、遅延時間と。データ集録を開始します。Matlab ソフトウェアは自動的にレーザー光ビーム レーザー露出オーバーを避けるために完了をブロックするシャッターを閉じるし、起動を渡すためにシャッターを開きます。 メモ: は、レーザーのパルス繰り返し周波数 (1 kHz) によって限定、256 × 256 ピクセルで画像のデータ収集を完了する約 1 分かかります。</li>
	<li>生データを処理し、ボリュームレンダ リング38を介して最大強度の投射 (MIP)13を 2 次元 (2 D) や 3 D PAM 画像を視覚化します。</li>
	<li>超音波探触子をアンマウント、脱イオン水を使用してヒントをすすぎ、それを収納ケースに入れてください。</li>
	<li>眼底カメラにウサギを転送し、再眼底を調べます。この手順は、イメージング セッション後眼底のすべての形態の変化があるかどうかを確認するのに役立ちます。</li>
	<li>角膜症、角膜表面の脱水を防ぐために実験中に点眼と角膜すべて 2 分を洗い流してください。監視および記録の動物きゅうしょ 15 分毎。 注: パム、OCT と眼底イメージング セッション約 1 時間かかります。</li>
</ol>6. 投稿画像
<ol><li>眼底再検査は眼底カメラを使用して後、V ゲルの場合は接続を切断します。目薬を使って目を洗う、デキサメサゾン眼軟膏ポリミキシン B 硫酸塩とネオマイシン フルルビプロフェン眼科を適用し、目を閉じます。</li>
	<li>回復室に水循環毛布でウサギを転送します。シールド ボックスの光から、ウサギが自然に目覚めるまで待ちます。この期間の間に動物きゅうしょ 15 分毎に監視を記録しておくし、記録の動物施設にコピーを返します。</li>
	<li>ウサギは目覚めるし、アクティブな警告、歩行は、通常、動物施設に戻る転送します。急性実験が予定されている場合は、安楽死ソリューションを使用して動物を安楽死させる (e.g。、Beuthanasia、0.22 mL/kg、耳静脈に静脈注射) の死体の処分。</li>
	<li>ソフトウェアとレーザーをオフにします。光学ベンチをクリーンアップします。</li>
</ol>

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著者が明らかに何もありません。

高品質眼底画像はそのままで、通常の涙映画が欠かせません。不規則で劣化した涙膜が大幅イメージの質42に低下します。涙液膜の整合性を維持し、角膜の表在性点状表層角膜症を防ぐため、2 分に約非常に頻繁に目薬を使用して角膜を潤滑するが重要です。目の不透明度に関する問題がないか、細隙灯を使用し、フルオレセイン角膜の状態をチェックするストリップします...

最近数十年は、生体光音響イメージング1,2,3,4,5,6,7 のフィールドの爆発的な発展を目撃しています。 ,8。光の音へのエネルギー変換に基づく、新興の光音響イメージング縮尺が細胞内小器官、細胞、組織、器官から小動物全身に生体試料を可視化できるし、その解剖を明らかにすることができます、機能、分子、遺伝子、代謝情報1,2,9,10、11,12。光音響イメージング細胞生物学13,14, 血管生物学15,16, 神経17,18 などのバイオ分野の範囲のユニークなアプリケーションを見つけた、腫瘍19,20,21,22、皮膚23、薬理24、および血液25,26。眼科、眼は、光音響応用イメージング科学者や臨床医から実質的な興味を集めている、現在アクティブな調査中です。
対照的に定期的に使用される眼撮像技術27(FA) 血管造影とインドシアニン グリーン造影 (ICGA) (蛍光のコントラストに基づく)、光干渉断層計 (OCT) など (光散乱コントラストに基づく)、派生 10 月血管造影 (赤い血球の反対の動きに基づく)、コントラスト メカニズムとして使用して光吸収イメージング音響眼。従来眼イメージング技術とは異なる、眼組織の28の病態生理学的状態に通常関連付けられている目の光吸収特性を研究するためのユニークなツールを提供します。までに、かなり優秀な作品は、光音響イメージング29,30,31,32,33,34,35、眼で行われています。 36,37, しかし、これらの研究はラット、マウス等の小動物の目の後方のセグメントに焦点を当てます。先駆的な研究はよく眼科で光音響イメージングの可能性を示すが、ラットおよびマウスの眼球サイズ以来技術の臨床的翻訳の方に行くには長い道のりはまだははるかに小さい (1-3 未満) より人間。かなり長距離超音波の伝搬、により画像大きく目の後方のセグメントの技術を使用する場合非常に信号強度と画質が苦しみます。
この目標に向かって私たちは最近非侵襲的、報告を使用して生きているウサギのラベル無料網脈絡膜イメージング統合光音響顕微鏡 (PAM) と周波数領域 10 月 (SD 10 月)38。システムは、優れたパフォーマンス、網膜と脈絡膜内在性吸収と眼組織の散乱コントラストに基づいてより大きい動物の目を可視化することができます。家兎における予備的な結果を示す PAM がレーザー照射線量を使用して個々 の網膜と脈絡膜血管を区別して非侵襲的 (~ 80 ニュージャージー州) アメリカ規格協会 (ANSI) 安全限度以下大幅 (160 ニュージャージー) 570 でnm39;OCT は明らかに強膜、脈絡膜、網膜の異なるレイヤーを解決できます。PAM を使用してより大きい動物の後部セグメント イメージングの非常に最初のデモンストレーションで、ウサギ (18.1 mm)40の眼球のサイズがの軸の長さのほぼ 80% であることを考慮した技術の臨床的翻訳の方の主要なステップがあります。人間 (23.9 mm)。
この作品ではデュアル モダリティ イメージング システム ・生活家兎における非侵襲的、ラベル無料網脈絡膜イメージング用実験プロトコルの詳細な説明を提供し、代表的な網膜を介してシステムのパフォーマンスを示すと脈絡膜の撮像結果。

<table border="0" cellpadding="0" cellspacing="0" style="width:824px;" width="822">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Dual-modality imaging system</td>
			<td style="width:279px;"></td>
			<td style="width:157px;"></td>
			<td style="width:159px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">OPO laser</td>
			<td style="width:279px;">Ekspla (Vilnius, Lithuania)</td>
			<td style="width:157px;">NT-242</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Beam attenuator</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">AHWP10M-600</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Motorized rotation stage</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">PRM1/MZ8</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Motorized rotation stage controller</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">TDC001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Focusing lens</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">AC254-250-B</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Pinhole</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">P50S</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Collimating lens</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">AC127-030-B</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Photodiode</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">PDA36A&#160;</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Laser shutter</td>
			<td style="width:279px;">Vincent Associates Inc. (Toronto, Canada)</td>
			<td style="width:157px;">LS6S2T0</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Laser shutter driver</td>
			<td style="width:279px;">Vincent Associates Inc. (Toronto, Canada)</td>
			<td style="width:157px;">VCM-D1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Dichroic mirror</td>
			<td style="width:279px;">Semrock, Inc. (Rochester, NY, USA)</td>
			<td style="width:157px;">Di03-R785-t3-25&#215;36</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Scan lens</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">OCT-LK3-BB</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Ophthalmic lens</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">AC080-010-B-ML</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Ultrasonic transducer</td>
			<td style="width:279px;">Optosonic Inc. (Arcadia, CA, USA)</td>
			<td style="width:157px;">Custom</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Amplifier</td>
			<td style="width:279px;">L3 Narda-MITEQ (Hauppauge, NY, USA)</td>
			<td style="width:157px;">AU-1647</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Band-pass filter</td>
			<td style="width:279px;">Mini-Circuits (Brooklyn, NY, USA)</td>
			<td style="width:157px;">BLP-30+</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Digitizer</td>
			<td style="width:279px;">DynamicSignals LLC (Lockport, IL, USA)</td>
			<td style="width:157px;">PX1500-4&#160;</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Synchronization electronics</td>
			<td style="width:279px;">National Instruments Corporation (Austin, TX, USA)</td>
			<td style="width:157px;">USB-6353</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">OCT module</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">Ganymede-II-HR</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Dispersion compensation glass</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">LSM03DC</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Illumination LED light</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">MCWHF2&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Power meter</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">S121C&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Power meter interface</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td>PM100USB&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Height measurement tool&#160;</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td>BHM1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Fundus camera</td>
			<td style="width:279px;">Topcon Corporation (Tokyo, Japan)&#160;</td>
			<td>TRC 50EX</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Matlab</td>
			<td style="width:279px;">MathWorks (Natick, MA, USA)</td>
			<td style="width:157px;">2017a</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Oscilloscope</td>
			<td style="width:279px;">Teledyne LeCroy (Chestnut Ridge, NY, USA)</td>
			<td style="width:157px;">WaveJet 354T</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Animal experiment</td>
			<td style="width:279px;"></td>
			<td style="width:157px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Water-circulating blanket</td>
			<td style="width:279px;">Stryker Corporation (Kalamazoo, MI, USA)</td>
			<td style="width:157px;">TP-700</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Ketamine hydrochloride injection</td>
			<td style="width:279px;">Par pharmaceutical, Inc. (Woodcliff Lake, NJ, USA)</td>
			<td style="width:157px;">NDC code 42023-115-10</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Xylazine hydrochloride</td>
			<td>VetOne (Boise, ID, USA)</td>
			<td style="width:157px;">NDC code 13985-704-10</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Tropicamide ophthalmic</td>
			<td style="width:279px;">Akorn Pharmaceuticals Inc. (Lake Forest, IL, USA)</td>
			<td style="width:157px;">NDC code 17478-102-12</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Phenylephrine hydrochloride ophthalmic</td>
			<td style="width:279px;">Paragon BioTeck, Inc. (Portland, OR, USA)</td>
			<td style="width:157px;">NDC code 42702-102-15</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Eye lubricant</td>
			<td style="width:279px;">Hub Pharmaceuticals LLC (Rancho Cucamonga, CA, USA)</td>
			<td style="width:157px;">NDC code 17238-610-15</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Eyewash</td>
			<td style="width:279px;">Altaire Pharmaceuticals, Inc. (Aquebogue, NY, USA)</td>
			<td style="width:157px;">NDC code 59390-175-18</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Tetracaine hydrochloride ophthalmic solution</td>
			<td style="width:279px;">Bausch &#38; Lomb, Inc. (Rochester, NY, USA)</td>
			<td style="width:157px;">NDC code 24208-920-64</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Flurbiprofen sodium ophthalmic solution</td>
			<td style="width:279px;">Bausch &#38; Lomb, Inc. (Rochester, NY, USA)</td>
			<td style="width:157px;">NDC code 24208-314-25</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:229px;">Neomycin and Polymyxin B Sulfates and Dexamethasone Ophthalmic Ointment</td>
			<td style="width:279px;">Bausch &#38; Lomb, Inc. (Rochester, NY, USA)</td>
			<td style="width:157px;">NDC code 24208-795-35</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Meloxicam injection</td>
			<td style="width:279px;">Henry Schein Inc. (Queens, NY, USA)</td>
			<td style="width:157px;">NDC code 11695-6925-1</td>
			<td></td>
		</tr>
	</tbody>
</table>

novel photoacoustic microscopy and optical coherence tomography dual-modality chorioretinal imaging in living rabbit eyes

音響眼球画像処理は、新たなイメージング技術音の波に光エネルギーを変換することによって眼の組織を視覚化できる非侵襲的、集中的な調査の下は、現在眼科です。ただし、ほとんどは、これまでの仕事は小さな眼球サイズによる臨床人間の翻訳のための課題がラットやマウスなどの小動物の目の後方のセグメントの画像に焦点を当てた報告。本稿では、新規光音響顕微鏡 (PAM) ウサギなどの大型動物の目の後部セグメント画像用光コヒーレンス断層法 (OCT) デュアル モダリティ システムをについて説明します。システム構成、システムの配置、動物の準備、およびin vivoウサギのイメージングの非侵襲的、ラベル無料網脈絡膜のデュアル モダリティ実験プロトコルの詳細します。PAM および OCT による網膜と脈絡膜の血管を含む、代表的な実験結果により、手法の有効性を実証する.この原稿は、ウサギの撮像結果の再現より大きい動物の光音響分光法眼を促進の実用的なガイドを提供します。

デュアル モダリティ イメージング システムと実験的なプロトコルは、筆者らの研究室では 4 つのニュージーランド白ウサギを用いた正常にテストされています。次は、典型的な結果をいくつか紹介します。
パムと SD 10 月デュアル モダリティの画像処理システムの概略図を図 1に示します。次のモ?...

Watch this Scientific Journal Video about Novel Photoacoustic Microscopy and Optical Coherence Tomography Dual-modality Chorioretinal Imaging in Living Rabbit Eyes at JoVE.com

Novel Photoacoustic Microscopy and Optical Coherence Tomography Dual-modality Chorioretinal Imaging in Living Rabbit Eyes

本稿では、新規のセットアップと光音響顕微鏡やウサギなどの大型動物の非侵襲的、ラベル無料網脈絡膜イメージング用光断層影像デュアル モダリティ装置の操作手順をについて説明します。

生活家兎眼に新規光音響顕微鏡、光コヒーレンストモグラフィーのデュアル モダリティ網脈絡膜イメージング

Photoacoustic ocular imaging is an emerging ophthalmic imaging technology that can noninvasively visualize ocular tissue by converting light energy into ...

novel-photoacoustic-microscopy-optical-coherence-tomography-dual

Research

JoVE Journal

Engineering

10.8K ビュー.  University of Michigan. 本稿では、新規のセットアップと光音響顕微鏡やウサギなどの大型動物の非侵襲的、ラベル無料網脈絡膜イメージング用光断層影像デュアル モダリティ装置の操作手順をについて説明します。

ビデオ: Novel Photoacoustic Microscopy and Optical Coherence Tomography Dual-modality Chorioretinal Imaging in Living Rabbit Eyes

Construction and Testing of Coin Cells of Lithium Ion Batteries

Rechargeable lithium ion batteries have wide applications in electronics, where customers always demand more capacity and longer lifetime. Lithium ion batteries have also been considered to be used in electric and hybrid vehicles1 or even electrical grid stabilization systems2. All these applications simulate a dramatic increase in the research and development of battery materials3-7, including new materials3,8, doping9, nanostructuring10-13, coatings or surface modifications14-17 and novel binders18. Consequently, an increasing number of physicists, chemists and materials scientists have recently ventured into this area. Coin cells are widely used in research laboratories to test new battery materials; even for the research and development that target large-scale and high-power applications, small coin cells are often used to test the capacities and rate capabilities of new materials in the initial stage. 
 In 2010, we started a National Science Foundation (NSF) sponsored research project to investigate the surface adsorption and disordering in battery materials (grant no. DMR-1006515). In the initial stage of this project, we have struggled to learn the techniques of assembling and testing coin cells, which cannot be achieved without numerous help of other researchers in other universities (through frequent calls, email exchanges and two site visits). Thus, we feel that it is beneficial to document, by both text and video, a protocol of assembling and testing a coin cell, which will help other new researchers in this field. This effort represents the "Broader Impact" activities of our NSF project, and it will also help to educate and inspire students.
In this video article, we document a protocol to assemble a CR2032 coin cell with a LiCoO2 working electrode, a Li counter electrode, and (the mostly commonly used) polyvinylidene fluoride (PVDF) binder. To ensure new learners to readily repeat the protocol, we keep the protocol as specific and explicit as we can. However, it is important to note that in specific research and development work, many parameters adopted here can be varied. First, one can make coin cells of different sizes and test the working electrode against a counter electrode other than Li. Second, the amounts of C black and binder added into the working electrodes are often varied to suit the particular purpose of research; for example, large amounts of C black or even inert powder were added to the working electrode to test the "intrinsic" performance of cathode materials14. Third, better binders (other than PVDF) have also developed and used18. Finally, other types of electrolytes (instead of LiPF6) can also be used; in fact, certain high-voltage electrode materials will require the uses of special electrolytes7.

A protocol to construct and test coin cells of lithium ion batteries is described. The specific procedures of making a working electrode, preparing a counter electrode, assembling a cell inside a glovebox and testing the cell are presented.

リチウムイオン電池のコイン細胞の構築とテスト

Rechargeable lithium ion batteries have wide applications in electronics, where customers always demand more capacity and longer lifetime.  Lithium ion ...

工学

Fabricating Metamaterials Using the Fiber Drawing Method

Metamaterials are man-made composite materials, fabricated by assembling components much smaller than the wavelength at which they operate 1. They owe their electromagnetic properties to the structure of their constituents, instead of the atoms that compose them. For example, sub-wavelength metal wires can be arranged to possess an effective electric permittivity that is either positive or negative at a given frequency, in contrast to the metals themselves 2. This unprecedented control over the behaviour of light can potentially lead to a number of novel devices, such as invisibility cloaks 3, negative refractive index materials 4, and lenses that resolve objects below the diffraction limit 5. However, metamaterials operating at optical, mid-infrared and terahertz frequencies are conventionally made using nano- and micro-fabrication techniques that are expensive and produce samples that are at most a few centimetres in size 6-7. Here we present a fabrication method to produce hundreds of meters of metal wire metamaterials in fiber form, which exhibit a terahertz plasmonic response 8. We combine the stack-and-draw technique used to produce microstructured polymer optical fiber 9 with the Taylor-wire process 10, using indium wires inside polymethylmethacrylate (PMMA) tubes. PMMA is chosen because it is an easy to handle, drawable dielectric with suitable optical properties in the terahertz region; indium because it has a melting temperature of 156.6 &deg;C which is appropriate for codrawing with PMMA. We include an indium wire of 1 mm diameter and 99.99% purity in a PMMA tube with 1 mm inner diameter (ID) and 12 mm outside diameter (OD) which is sealed at one end. The tube is evacuated and drawn down to an outer diameter of 1.2 mm. The resulting fiber is then cut into smaller pieces, and stacked into a larger PMMA tube. This stack is sealed at one end and fed into a furnace while being rapidly drawn, reducing the diameter of the structure by a factor of 10, and increasing the length by a factor of 100. Such fibers possess features on the micro- and nano- scale, are inherently flexible, mass-producible, and can be woven to exhibit electromagnetic properties that are not found in nature. They represent a promising platform for a number of novel devices from terahertz to optical frequencies, such as invisible fibers, woven negative refractive index cloths, and super-resolving lenses.

Metamaterials at terahertz frequencies offer unique opportunities, but are challenging to fabricate in bulk. We adapt the fabrication procedure for microstructured polymer optical fibers to inexpensively fabricate metamaterials potentially on an industrial scale. We produce polymethylmethacrylate fibers containing ~10 &mu;m diameter indium wires separated by ~100 &mu;m, which exhibit a terahertz plasmonic response.

ファイバー描画法を用いたメタマテリアルの作製

Metamaterials are man-made composite materials, fabricated by assembling components much smaller than the wavelength at which they operate 1. They owe ...

Measurement of Coherence Decay in GaMnAs Using Femtosecond Four-wave Mixing

The application of femtosecond four-wave mixing to the study of fundamental properties of diluted magnetic semiconductors ((s,p)-d hybridization, spin-flip scattering) is described, using experiments on GaMnAs as a prototype III-Mn-V system.&#160; Spectrally-resolved and time-resolved experimental configurations are described, including the use of zero-background autocorrelation techniques for pulse optimization.&#160; The etching process used to prepare GaMnAs samples for four-wave mixing experiments is also highlighted.&#160; The high temporal resolution of this technique, afforded by the use of short (20 fsec) optical pulses, permits the rapid spin-flip scattering process in this system to be studied directly in the time domain, providing new insight into the strong exchange coupling responsible for carrier-mediated ferromagnetism.&#160; We also show that spectral resolution of the four-wave mixing signal allows one to extract clear signatures of (s,p)-d hybridization in this system, unlike linear spectroscopy techniques.&#160;&#160; This increased sensitivity is due to the nonlinearity of the technique, which suppresses defect-related contributions to the optical response. This method may be used to measure the time scale for coherence decay (tied to the fastest scattering processes) in a wide variety of semiconductor systems of interest for next generation electronics and optoelectronics.

The technique of femtosecond four-wave mixing is described, including spectrally-resolved and time-resolved configurations. We illustrate the utility of this technique for the investigation of crucial physical properties in the III-V diluted magnetic semiconductors, afforded by its nonlinearity and high temporal resolution.

フェムト秒四光波混合を用いてのGaMnAsコヒーレンス崩壊の測定

The application of femtosecond four-wave mixing to the study of fundamental properties of diluted magnetic semiconductors ((s,p)-d hybridization, ...

Coherent anti-Stokes Raman Scattering (CARS) Microscopy Visualizes Pharmaceutical Tablets During Dissolution

Traditional pharmaceutical dissolution tests determine the amount of drug dissolved over time by measuring drug content in the dissolution medium. This method provides little direct information about what is happening on the surface of the dissolving tablet. As the tablet surface composition and structure can change during dissolution, it is essential to monitor it during dissolution testing. In this work coherent anti-Stokes Raman scattering microscopy is used to image the surface of tablets during dissolution while UV absorption spectroscopy is simultaneously providing inline analysis of dissolved drug concentration for tablets containing a 50% mixture of theophylline anhydrate and ethyl cellulose. The measurements showed that in situ CARS microscopy is capable of imaging selectively theophylline in the presence of ethyl cellulose. Additionally, the theophylline anhydrate converted to theophylline monohydrate during dissolution, with needle-shaped crystals growing on the tablet surface during dissolution. The conversion of theophylline anhydrate to monohydrate, combined with reduced exposure of the drug to the flowing dissolution medium resulted in decreased dissolution rates. Our results show that in situ CARS microscopy combined with inline UV absorption spectroscopy is capable of monitoring pharmaceutical tablet dissolution and correlating surface changes with changes in dissolution rate.

Coherent anti-Stokes Raman Scattering CARS Microscopy Visualizes Pharmaceutical Tablets During Dissolution

Coherent anti-Stokes Raman scattering (CARS) microscopy is combined with an intrinsic flow-through dissolution setup to allow in situ and real-time visualization of the surface of pharmaceutical tablets undergoing dissolution. Using this custom-built setup, it is possible to correlate CARS videos with drug dissolution profiles recorded using inline UV absorption spectroscopy.

溶解中にコヒーレント反ストークスラマン散乱（CARS）顕微鏡可視化薬錠

Traditional pharmaceutical dissolution tests determine the amount of drug dissolved over time by measuring drug content in the dissolution medium. This ...

Writing Bragg Gratings in Multicore Fibers

Fiber Bragg gratings in multicore fibers can be used as compact and robust filters in astronomical and other research and commercial applications. Strong suppression at a single wavelength requires that all cores have matching transmission profiles. These gratings cannot be inscribed using the same method as for single-core fibers because the curved surface of the cladding acts as a lens, focusing the incoming UV laser beam and causing variations in exposure between cores. Therefore we use an additional optical element to ensure that the beam shape does not change while passing through the cross-section of the multicore fiber. This consists of a glass capillary tube which has been polished flat on one side, which is then placed over the section of the fiber to be inscribed. The laser beam enters the fiber through the flat surface of the capillary tube and hence maintains its original dimensions. This paper demonstrates the improvements in core-to-core uniformity for a 7-core fiber using this method. The technique can be generalized to larger multicore fibers.

We describe a technique for inscribing identical fiber Bragg gratings into each core of a multicore fiber. This is achieved by introducing an additional surface into the optical path to mitigate lensing by the curved surface of the fiber cladding.

マルチコアファイバーブラッググレーティングを書きます

Fiber Bragg gratings in multicore fibers can be used as compact and robust filters in astronomical and other research and commercial applications. Strong ...

Compact Lens-less Digital Holographic Microscope for MEMS Inspection and Characterization

A micro-electro-mechanical-system (MEMS) is a widely used component in many industries, including energy, biotechnology, medical, communications, and automotive. However, effective inspection and characterization metrology systems are needed to ensure the functional reliability of MEMS. This study presents a system based on digital holography as a tool for MEMS metrology. Digital holography has gained increasing attention in the past 20 years. With the fast development and decreasing cost of sensor arrays, resolution of such systems has increased broadening potential applications. Thus, it has attracted attention from both research and industry sides as a potential reliable tool for industrial metrology. Indeed, by recording the interference pattern between an object beam (which contains sample height information) and a reference beam on a CCD camera, one can retrieve the quantitative phase information of an object. However, most of digital holographic systems are bulky and thus not easy to implement on industry production lines. The novelty of the system presented is that it is lens-less and thus very compact. In this study, it is shown that the Compact Digital Holographic Microscope (CDHM) can be used to evaluate several characteristics typically consider as criteria in MEMS inspections. The surface profiles of MEMS in both static and dynamic conditions are presented. Comparison with AFM is investigated to validate the accuracy of the CDHM.

We present a compact reflection digital holographic system (CDHM) for inspection and characterization of MEMS devices. A lens-less design using a diverging input wave providing natural geometrical magnification is demonstrated. Both static and dynamic studies are presented.

MEMS検査および特性評価のためのコンパクトなレンズレスデジタルホログラフィック顕微鏡

A micro-electro-mechanical-system (MEMS) is a widely used component in many industries, including energy, biotechnology, medical, communications, and ...

Dynamic Pore-scale Reservoir-condition Imaging of Reaction in Carbonates Using Synchrotron Fast Tomography

Underground storage permanence is a major concern for carbon capture and storage. Pumping CO2 into carbonate reservoirs has the potential to dissolve geologic seals and allow CO2 to escape. However, the dissolution processes at reservoir conditions are poorly understood. Thus, time-resolved experiments are needed to observe and predict the nature and rate of dissolution at the pore scale. Synchrotron fast tomography is a method of taking high-resolution time-resolved images of complex pore structures much more quickly than traditional &#181;-CT. The Diamond Lightsource Pink Beam was used to dynamically image dissolution of limestone in the presence of CO2-saturated brine at reservoir conditions. 100 scans were taken at a 6.1 &#181;m resolution over a period of 2 hours. The images were segmented and the porosity and permeability were measured using image analysis and network extraction. Porosity increased uniformly along the length of the sample; however, the rate of increase of both porosity and permeability slowed at later times.

Synchrotron fast tomography was used to dynamically image dissolution of limestone in the presence of CO2-saturated brine at reservoir conditions. 100 scans were taken at a 6.1 &#181;m resolution over a period of 2 h.

シンクロトロン高速断層撮影法を使用した炭酸における反応の動的細孔スケール貯水池条件イメージング

Underground storage permanence is a major concern for carbon capture and storage. Pumping CO2 into carbonate reservoirs has the potential to dissolve ...

In Depth Analyses of LEDs by a Combination of X-ray Computed Tomography (CT) and Light Microscopy (LM) Correlated with Scanning Electron Microscopy (SEM)

In failure analysis, device characterization and reverse engineering of light emitting diodes (LEDs), and similar electronic components of micro-characterization, plays an important role. Commonly, different techniques like X-ray computed tomography (CT), light microscopy (LM) and scanning electron microscopy (SEM) are used separately. Similarly, the results have to be treated for each technique independently. Here a comprehensive study is shown which demonstrates the potentials leveraged by linking CT, LM and SEM. In depth characterization is performed on a white emitting LED, which can be operated throughout all characterization steps. Major advantages are: planned preparation of defined cross sections, correlation of optical properties to structural and compositional information, as well as reliable identification of different functional regions. This results from the breadth of information available from identical regions of interest (ROIs): polarization contrast, bright and dark-field LM images, as well as optical images of the LED cross section in operation. This is supplemented by SEM imaging techniques and micro-analysis using energy dispersive X-ray spectroscopy.

In Depth Analyses of LEDs by a Combination of X-ray Computed Tomography CT and Light Microscopy LM Correlated with Scanning Electron Microscopy SEM

A workflow for comprehensive micro-characterization of active optical devices is outlined. It contains structural as well as functional investigations by means of CT, LM and SEM. The method is demonstrated for a white LED which can be still be operated during characterization.

走査型電子顕微鏡（SEM）と相関X線コンピュータ断層撮影（CT）および光学顕微鏡（LM）の組合せによるLEDの深さ分析で

In failure analysis, device characterization and reverse engineering of light emitting diodes (LEDs), and similar electronic components of ...

Optical Trap Loading of Dielectric Microparticles In Air

We demonstrate a method to trap a selected dielectric microparticle in air using radiation pressure from a single-beam gradient optical trap. Randomly scattered dielectric microparticles adhered to a glass substrate are momentarily detached using ultrasonic vibrations generated by a piezoelectric transducer (PZT). Then, the optical beam focused on a selected particle lifts it up to the optical trap while the vibrationally excited microparticles fall back to the substrate. A particle may be trapped at the nominal focus of the trapping beam or at a position above the focus (referred to here as the levitation position) where gravity provides the restoring force. After the measurement, the trapped particle can be placed at a desired position on the substrate in a controlled manner. 
In this protocol, an experimental procedure for selective optical trap loading in air is outlined. First, the experimental setup is briefly introduced. Second, the design and fabrication of a PZT holder and a sample enclosure are illustrated in detail. The optical trap loading of a selected microparticle is then demonstrated with step-by-step instructions including sample preparation, launching into the trap, and use of electrostatic force to excite particle motion in the trap and measure charge. Finally, we present recorded particle trajectories of Brownian and ballistic motions of a trapped microparticle in air. These trajectories can be used to measure stiffness or to verify optical alignment through time domain and frequency domain analysis. Selective trap loading enables optical tweezers to track a particle and its changes over repeated trap loadings in a reversible manner, thereby enabling studies of particle-surface interaction.

A protocol for launching and stably trapping selected dielectric microparticles in air is presented.

空気中の誘電体微粒子の光学トラップのロード

We demonstrate a method to trap a selected dielectric microparticle in air using radiation pressure from a single-beam gradient optical trap. Randomly ...

Microfluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy (ATOM)

Scaling the number of measurable parameters, which allows for multidimensional data analysis and thus higher-confidence statistical results, has been the main trend in the advanced development of flow cytometry. Notably, adding high-resolution imaging capabilities allows for the complex morphological analysis of cellular/sub-cellular structures. This is not possible with standard flow cytometers. However, it is valuable for advancing our knowledge of cellular functions and can benefit life science research, clinical diagnostics, and environmental monitoring. Incorporating imaging capabilities into flow cytometry compromises the assay throughput, primarily due to the limitations on speed and sensitivity in the camera technologies. To overcome this speed or throughput challenge facing imaging flow cytometry while preserving the image quality, asymmetric-detection time-stretch optical microscopy (ATOM) has been demonstrated to enable high-contrast, single-cell imaging with sub-cellular resolution, at an imaging throughput as high as 100,000 cells/s. Based on the imaging concept of conventional time-stretch imaging, which relies on all-optical image encoding and retrieval through the use of ultrafast broadband laser pulses, ATOM further advances imaging performance by enhancing the image contrast of unlabeled/unstained cells. This is achieved by accessing the phase-gradient information of the cells, which is spectrally encoded into single-shot broadband pulses. Hence, ATOM is particularly advantageous in high-throughput measurements of single-cell morphology and texture &#8211; information indicative of cell types, states, and even functions. Ultimately, this could become a powerful imaging flow cytometry platform for the biophysical phenotyping of cells, complementing the current state-of-the-art biochemical-marker-based cellular assay. This work describes a protocol to establish the key modules of an ATOM system (from optical frontend to data processing and visualization backend), as well as the workflow of imaging flow cytometry based on ATOM, using human cells and micro-algae as the examples.

Microfluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy ATOM

This protocol describes the implementation of an asymmetric-detection time-stretch optical microscopy system for single-cell imaging in ultrafast microfluidic flow and its applications in imaging flow cytometry.

非対称検出時間ストレッチ光学顕微鏡（ATOM）によるマイクロ流体イメージングフローサイトメトリー

Scaling the number of measurable parameters, which allows for multidimensional data analysis and thus higher-confidence statistical results, has been the ...

生活家兎眼に新規光音響顕微鏡、光コヒーレンストモグラフィーのデュアル モダリティ網脈絡膜イメージング

生活家兎眼に新規光音響顕微鏡、光コヒーレンストモグラフィーのデュアルモダリティ網脈絡膜イメージング