彼らの親切なサポートの災害医学、外傷学 (KMC) スウェーデン防衛庁 (FOI) センターのリンシェーピングのスタッフの方々 に感謝したいと思います。

ここで説明した研究は、ローカル倫理ガイドラインに従った、リンシェーピング (許可番号 2015/99-31) の地域の倫理審査委員会によって承認されました。
1. インフォームド コンセントとスクリーニング
<ol><li>サブジェクトからインフォームド コンセントを取得します。</li>
	<li>選択/除外基準によると画面します。 注: 選択基準であった: (i) 健康な成人 > 18 歳の (ii) (iii) の筆記と口頭の情報を理解することは、口頭および書面による同意を提供します。除外基準であった: (i) できませんまたはインフォームド コンセント、(ii) 進行中の皮膚の状態、(iii) 心血管疾患、血管機能に干渉することができます (iv) 薬を提供するために不本意です。経口避妊薬が許可されます。件名は、このする必要があります経口避妊薬を使用している場合件名のプロトコル ファイル、(v) 瘢痕に記載されている無料と非打撲皮膚の額、(vi) のカフェインやタバコのテスト、および (vii) の発症する前に 2 h のない摂取量ない精力的な身体活動の少なくとも 2 h prテストの発症に ior。</li>
</ol>2. 順化、機器のセットアップ
<ol><li>少なくとも 20 分テストの発症する前に仰臥位で慣らすテスト対象をしましょう。 注: 正確な被写体の位置決めと装置は補足図 1 に示します。</li>
	<li>
(材料の表を参照してください) 指定されたソフトウェアとデジタル カメラを用いた毛細血管リフィル テストのカメラの記録を設定します。
	<ol>
<li>カメラとコンピューターに USB ケーブルを接続し、ビデオモードにカメラの設定を調整します。</li>
		<li>カメラをオンに、それはコンピューターに接続するかどうかを確認します。</li>
		<li>カメラのリモート制御ソフトウェアを起動 (材料の表を参照してください)。</li>
		<li>周囲の光を切り、測定エリアからコンピューターの画面を有効にします。</li>
	</ol>
</li>
</ol>3. データ集録
<ol><li>「人生の表示」をクリックして (Sic) ボタンを押し「スタート」.</li>
	<li>'Video' モードとラジオ ボタンをクリックして '事業' モードの切り替え (のみ、記録には影響しません、オンスクリーン プレゼンテーション)。被験者の額の生きているビデオ ストリームがカメラのディスプレイに、コンピューターの画面に表示されていることを確認します。</li>
	<li>被験者の額の真上約 15 cm の高さにカメラを調整します。</li>
	<li>カメラのフォーカスが"AF"(オート フォーカス) に設定されていることを確認します。</li>
	<li>テストの期間のためにまだ彼/彼女の頭を維持して控えに件名を求めます。言って、テスト中に閉じた彼/彼女の目を維持することが [ok] 件名を知らせる:「この枕上頭を配置して、テスト期間ためにまだあなたの頭を維持しますください。テストは最後の 20 秒間に話をしないでください。光を見つける場合に全体の時間のため閉鎖あなたの目を保つことができるあまりにも明るい」。</li>
	<li>録音しながらカメラを動かさないでください。</li>
	<li>「人生の表示」ウィンドウを見つけるし、「HD 記録」をクリックして記録 (秒) の時間、カメラのディスプレイに見ることができる 20 の s. 注意の連続録音を開始するには最初の 5 s 記録の額の通常の毛細血管の血液濃度を取得することです。</li>
	<li>5 の終わりに記録、ベースラインの適用圧力同等化および温度の絶縁材のための 5 s. 使用指と額の間プラスチック小さじと人差し指の測定領域に圧力をブランチング事務所。</li>
	<li>圧力を解放し、測定エリアから審査官の指とプラスチック製のティー スプーンをすぐに撤回します。</li>
	<li>別の 10 の記録を継続後の充血している応答を取得する s。</li>
	<li>「人生表示」ウィンドウで「停止」をクリックして録音を停止します。</li>
	<li>記録を停止した、省 (.mov 形式) でファイルに名前を付けるのため、ウィンドウが開きますビデオの名前、フォルダーを選択します。ビデオをコンピューター上の指定フォルダーにダウンロードしながら、コンピューターからカメラを取り外さないでください。</li>
</ol>4. データ解析
<ol><li>専用解析ソフトウェアで画像解析と曲線の工事を行うに進みます (材料の表を参照してください)。</li>
	<li>解析ソフトウェアをデスクトップのアイコンをダブルクリックして起動します。</li>
	<li>メイン画面で、ソフトウェアを開始しました「ムービー ページ」を選択します。</li>
	<li>「ムービー ページ」ウィンドウで、"ムービーを読み込む」をクリックします。ソフトウェアをアップロードし、映画を分析するを待ちます。これはビデオ ファイルのサイズによっては数分をかかる場合があります。</li>
	<li>アップロードと映画の分析が終了したら、一度「画像として保存」をクリックし、、ビデオを分析するソフトウェアを待ちます。 注: これは自動的に、個別の jpeg 画像に分かれてビデオ シーケンスを含むコンピューターに新しいフォルダーが作成されます。</li>
	<li>ビデオの個々 の jpeg 画像分割が完了すると、検索し、メイン画面のウィンドウで「作物画像ページ」を選択。</li>
	<li>「作物の画像のページ」に「最初の写真」をクリックしてビデオから jpeg 画像を含むフォルダーに移動し、を分析するのには、最初の写真を選択ソフトウェアは、そのバッチの次の写真を自動的に選択されます。</li>
	<li>「編集」メニューをクリックし、関心領域の円形または四角形のマーカーの間を選択します。</li>
	<li>「写真を選択」ウィンドウで測定エリアから調査官の指が完全に取り消されるバッチの「実績写真」ボックスと最初の写真の名前を入力を検索します。この写真を検索するには、ビデオから個々 の写真が含まれるフォルダーを参照します。</li>
	<li>クリックしてマウスの左ボタンを押したまま関心領域を選択し、湯通しの領域の境界内にあるフォト ウィンドウの円または四角形を描画します。選択した関心領域は、バッチのすべての写真に自動的に適用されます。</li>
	<li>「ファイル」メニューをクリックし、今後の参考のための興味の選択した領域の場所を保存する「ROI の保存」を選択します。</li>
	<li>「曲線のトラッカー」ボタンをクリックし、関心領域を分析するソフトウェアを待ちます。</li>
	<li>曲線の追跡ツール ウィンドウが開き、テストの持続期間のための興味の選択した領域の赤血球濃度の変化が曲線として表示されます。別のラジオ ボタンをクリックして画面上のプレゼンテーションを変更できます。これは生のデータには影響しません。</li>
	<li>「データのエクスポート」をクリックし、、さらに分析生データのトラン スクリプトを保存します。</li>
	<li>生データのトラン スクリプトで移動という名前の列に「意味強度 TiVi 値」「合計時間」と「毛細血管再充満時間」1 から始まる「画像番号」のこの列の横ある 3 つの新しい列を生成 s 0 s および 0 s と 1 の増分 0.02 の s、および 0.02 s、それぞれ。 注:「キャピラリー詰替時間」ブランチング圧力のリリースに後続の反応時間と相関関係し、測定領域から捜査員の指が完全に取り消される最初の写真を表す平均値から始まります。</li>
	<li>ブランチングの演習中に捜査員の人差し指とプラスチック製のティー スプーンによって生成されるノイズを含んでいるこれらだけベースラインと毛細血管再充満時間の値を削除します。</li>
	<li>ベースライン測定値の平均値を計算 (最初の 5 s の記録)。</li>
	<li>最初の値と等しいか、計算された平均ベースライン値とメモ「キャピラリー補充時間」列の時刻を上回るを見つけます。この時点でポイントを表す「基準 1 に戻る時間」。</li>
	<li>最高圧力解放後に生成される値の強度の値を意味して「毛細血管リフィルひととき」欄の時間ポイントを見つけます。この時点でポイントを表す「ピーク時間」。</li>
	<li>検索の 2 番目の値は「毛細血管リフィル時間」列でベースラインの測定と注意時点の計算された平均値以下です。この時点でポイントを表す「基準 2 に戻る時間」。</li>
</ol>

<ol>
	<li>Strehlow, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+identification+of+shock+in+critically+ill+patients.">Early identification of shock in critically ill patients.</a> Emerg Med Clin North Am. 28 (1), 57-66 (2010).</li><li>Advanced Life Support, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Acute Medical Emergencies. , 67-84 (2010).</li><li>Brasel, K. J., Guse, C., Gentilello, L. M., Nirula, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heart+rate:+is+it+truly+a+vital+sign?.">Heart rate: is it truly a vital sign?.</a> J Trauma. 62 (4), 812-817 (2007).</li><li>De Backer, D., Ortiz, J. A., Salgado, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coupling+microcirculation+to+systemic+hemodynamics.">Coupling microcirculation to systemic hemodynamics.</a> Curr Opin Crit Care. 16 (3), 250-254 (2010).</li><li>Mrgan, M., Rytter, D., Brabrand, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Capillary+refill+time+is+a+predictor+of+short-term+mortality+for+adult+patients+admitted+to+a+medical+department:+an+observational+cohort+study.">Capillary refill time is a predictor of short-term mortality for adult patients admitted to a medical department: an observational cohort study.</a> Emerg Med J. 31 (12), 954-958 (2014).</li><li>Advanced Life Support, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Acute Medical Emergencies. , 55-65 (2010).</li><li>Lewin, J., Maconochie, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Capillary+refill+time+in+adults.">Capillary refill time in adults.</a> Emerg Med J. 25 (6), 325-326 (2008).</li><li>Lobos, A. T., Menon, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+multidisciplinary+survey+on+capillary+refill+time:+Inconsistent+performance+and+interpretation+of+a+common+clinical+test.">A multidisciplinary survey on capillary refill time: Inconsistent performance and interpretation of a common clinical test.</a> Pediatr Crit Care Med. 9 (4), 386-391 (2008).</li><li>Otieno, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Are+bedside+features+of+shock+reproducible+between+different+observers?.">Are bedside features of shock reproducible between different observers?.</a> Arch Dis Child. 89 (10), 977-979 (2004).</li><li>Raju, N. V., Maisels, M. J., Kring, E., Schwarz-Warner, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Capillary+refill+time+in+the+hands+and+feet+of+normal+newborn+infants.">Capillary refill time in the hands and feet of normal newborn infants.</a> Clin Pediatr (Phila). 38 (3), 139-144 (1999).</li><li>Klupp, N. L., Keenan, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+evaluation+of+the+reliability+and+validity+of+capillary+refill+time+test.">An evaluation of the reliability and validity of capillary refill time test.</a> The Foot. 17, 15-20 (2007).</li><li>Gorelick, M. H., Shaw, K. N., Baker, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+ambient+temperature+on+capillary+refill+in+healthy+children.">Effect of ambient temperature on capillary refill in healthy children.</a> Pediatrics. 92 (5), 699-702 (1993).</li><li>Schriger, D. L., Baraff, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Defining+normal+capillary+refill:+variation+with+age,+sex,+and+temperature.">Defining normal capillary refill: variation with age, sex, and temperature.</a> Ann Emerg Med. 17 (9), 932-935 (1988).</li><li>Pandey, A., John, B. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Capillary+refill+time.+Is+it+time+to+fill+the+gaps?.">Capillary refill time. Is it time to fill the gaps?.</a> Med J Armed Forces India. 69 (1), 97-98 (2013).</li><li>Strozik, K. S., Pieper, C. H., Roller, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Capillary+refilling+time+in+newborn+babies:+normal+values.">Capillary refilling time in newborn babies: normal values.</a> Arch Dis Child Fetal Neonatal Ed. 76 (3), F193-F196 (1997).</li><li>Doherty, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sub-epidermal+imaging+using+polarized+light+spectroscopy+for+assessment+of+skin+microcirculation.">Sub-epidermal imaging using polarized light spectroscopy for assessment of skin microcirculation.</a> Skin Res Technol. 13 (4), 472-484 (2007).</li><li>Noguchi, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+caffeine+contained+in+a+cup+of+coffee+on+microvascular+function+in+healthy+subjects.">Effect of caffeine contained in a cup of coffee on microvascular function in healthy subjects.</a> J Pharmacol Sci. 127 (2), 217-222 (2015).</li><li>Reus, W. F., Robson, M. C., Zachary, L., Heggers, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+effects+of+tobacco+smoking+on+blood+flow+in+the+cutaneous+micro-circulation.">Acute effects of tobacco smoking on blood flow in the cutaneous micro-circulation.</a> Br J Plast Surg. 37 (2), 213-215 (1984).</li><li>Lenasi, H., Strucl, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+regular+physical+training+on+cutaneous+microvascular+reactivity.">Effect of regular physical training on cutaneous microvascular reactivity.</a> Med Sci Sports Exerc. 36 (4), 606-612 (2004).</li><li>Anderson, B., Kelly, A. M., Kerr, D., Clooney, M., Jolley, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+patient+and+environmental+factors+on+capillary+refill+time+in+adults.">Impact of patient and environmental factors on capillary refill time in adults.</a> Am J Emerg Med. 26 (1), 62-65 (2008).</li><li>John, R. T., Henricson, J., Nilsson, G. E., Wilhelms, D., Anderson, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reflectance+spectroscopy:+to+shed+new+light+on+the+capillary+refill+test.">Reflectance spectroscopy: to shed new light on the capillary refill test.</a> J Biophotonics. , (2017).</li><li>Blaxter, L. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+automated+quasi-continuous+capillary+refill+timing+device.">An automated quasi-continuous capillary refill timing device.</a> Physiological measurement. 37, 83-99 (2016).</li><li>Bordoley, A., Irwin, R., Kalyani, V., MacDonald, C., Standish, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> D1GIT: Automated, temperature-calibrated measurement of capillary refill time. , (2012).</li><li>Kviesis-Kipge, E., Curkste, E., Spigulis, J., Eihvalde, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Real-time+analysis+of+skin+capillary-refill+processes+using+blue+LED.">Real-time analysis of skin capillary-refill processes using blue LED.</a> Proc. of SPIE. 771523, (2010).</li><li>Shavit, I., Brant, R., Nijssen-Jordan, C., Galbraith, R., Johnson, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+imaging+technique+to+measure+capillary-refill+time:+improving+diagnostic+accuracy+for+dehydration+in+young+children+with+gastroenteritis.">A novel imaging technique to measure capillary-refill time: improving diagnostic accuracy for dehydration in young children with gastroenteritis.</a> Pediatrics. 118 (6), 2402-2408 (2006).</li><li>Cohen, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monitor+to+Measure+Dermal+Blood+Flow+in+Critically+Ill+Patients:+Study.">Monitor to Measure Dermal Blood Flow in Critically Ill Patients: Study.</a> International Scholarly Research Notices. 578316, e578316 (2013).</li></ol>

試験の実施に関与していた WheelsBridge AB からの財政支援はありません。著者 JH エステルイェータランド県評議会によって採用しますが、WheelsBridge 株式会社とロイヤリティ契約CDA の年長の著者は WheelsBridge ab 級でフルタイムの学術の位置も限られた関与

システムで最良の結果を得るために環境要因による変動を制御する必要があります。すべての周囲の光をオフにする必要があります。計測領域の垂直方向の配置でカメラを配置する必要があります。定数測定領域を確保するために被験者の移動や測定中に話をしないで。同じ理由から、カメラはできれば動きを避けるために、測定領域に一定の距離を維持するためにスタンドにマウントされ?...

評価および古典的なバイタル サイン血圧 (BP)、心拍数 (HR)、呼吸 (RR)、酸素飽和度、ボディ温度1の批判的に病気の患者センターの選別。これらのパラメーターの変化は、循環の悪化の過程で比較的遅く表示されます。例えば、出血、出血量が中等度から重度の2、心拍増加に小文字を区別しないと非特異的マーカー3することができますまで BP の低下は発生しません。
毛細血管リフィル (CR テスト) は、詰替時間はバイタル サインとして冷たい、じめじめした、まだらにされた皮1,4の臨床出現前に変更と考えられる初期の循環虚脱の以前の適応があります。,5します。 アプリケーションによって通常毛細血管リフィル テストが実行され、離します会社ブランチングの湯通しの領域への血のリターンの秒単位でタイミングで皮膚に圧力。ガイドラインによると、通常健康な仰臥位患者6(例えば医師の指で) 会社の圧力の 5 秒のリリース後 2 秒以内詰替が行われます。テストのための理論的根拠は遅い詰替時間が悪い皮膚灌流、可能な敗血症、出血、急性心不全、低体温症などの重大なイベントの数の 1 つによって引き起こされるを示すでしょう。
現時点では、CR テスト6,7,8,9,10を実行するための最先端の方法に関するコンセンサスはありません。争点は、実際にブランチングやると主観的な依存の標準化の欠如 (すなわち肉眼) 詰替エンドポイント7,9,11の評価。さらに、性別影響 CR 時間12,13している兆候があります。温度、周囲温度と皮膚毛細血管再充満時間に影響を与える知られているが、どの程度まで明確ではありません。最後に、異なる測定サイト、周辺機器または、セントラルの使用はおそらくこのエリア14,15でいくつかの研究の結果の可変性のさらなる原因です。
現在の仕事で、血と CR テスト中に見られるその後の充血している応答の戻り値のコースを記録する光工学システムを使いました。システムでは、拡散反射光の偏光分光定量化可能な肉眼と時間、毛細血管リフィルのコースよりも詳細に記述して利用しています。システムには、92 の白色 Led と特別に開発されたソフトウェアの外部光リング装備標準的なデジタル カメラが装備されています。レンズと Led の前で直交接続 2 つの偏光フィルターに直接カメラに到達する組織における分極になる唯一の光をできるように皮膚の表面から反映されて光をブロックします。これは 0.5 mm 程度の深さに組織の「サブ表皮」イメージを生成します。その色面に画像を分割し、ピクセルごとに赤と緑のコンテンツを計算すると、赤血球の16の組織濃度に対応する値を生成します。システムの時間分解能は、ビデオ モードで、0.02 秒。

<table>
	<tbody>
		<tr>
			<td>TiVi701 Camera</td>
			<td>WheelsBridge AB</td>
			<td>TiVi701 Camera, version 1.5.1</td>
			<td>Software</td>
		</tr>
		<tr>
			<td>TiVi700</td>
			<td>WheelsBridge AB</td>
			<td>TiVi700 Analysis, version 1.2.9</td>
			<td>Software</td>
		</tr>
		<tr>
			<td>Canon EOS 700D</td>
			<td>Canon U.S.A., inc.</td>
			<td>Canon EOS 700D</td>
			<td>Digital SLR Camera</td>
		</tr>
		<tr>
			<td>Camera stand</td>
			<td>Manfrotto</td>
			<td>681B</td>
			<td>Modified camera stand to hold the digital camera in position</td>
		</tr>
		<tr>
			<td>Camera stand</td>
			<td>Disa Denmark</td>
			<td>9020B</td>
			<td>Modified camera stand to hold the digital camera in position</td>
		</tr>
	</tbody>
</table>

diffuse reflectance spectroscopy: getting the capillary refill test under one's thumb

毛細血管リフィル テストは、批判的に病気の患者の循環動態を見積もるために、1947 年に導入されました。その詰替 2 内発生状態の一般的ガイドライン 5 を解放した後 s 通常の健康的な仰臥位患者 (例えば医師の指で) 会社の圧力の s。遅い詰替時刻は、敗血症、血損失、低灌流、低体温症などの条件によって発生することができます悪い皮膚の血流を示します。導入以来、テストの臨床的有用性が議論されています。擁護派はその実現性とシンプルさを指摘し、心拍数などのバイタル サインの変化よりも早く血管の状態の変化を示すことができることを主張します。批評家は、その一方で、テストの実行方法の標準化の欠如と環境要因」、テストの感受性と同様、肉眼評価の非常に主観的な性質は著しく臨床的価値を下げることを強調します。本研究の目的は、詰替イベントのコースの詳細に説明し、潜在的に多くの目的を示唆して毛細血管の正確なエンドポイント値補充テスト拡散反射光の偏光分光法を用いたします。

キャピラリーの補充のコースを撮影では肉眼の評価を取得することはできませんデータの膨大な量を生成します。私達は、ここでさらに循環動態の悪化の早期指標として CR テストの有用性を改善するために新しいエンドポイントを提案します。我々 はこれらのエンドポイントを呼び出す:「ベースライン」、「血ゼロ」(または"BZ")"でも「ピーク時間」または"Tpk."ベー...

Watch this Scientific Journal Video about Diffuse Reflectance Spectroscopy: Getting the Capillary Refill Test Under One's Thumb at JoVE.com

Diffuse Reflectance Spectroscopy: Getting the Capillary Refill Test Under One's Thumb

このプロトコルでは、拡散反射光の偏光分光学の使用が毛細血管リフィル テストの臨床的有用性を向上する方法について説明します。拡散反射率分光法ビデオと新しい情報学的エンドポイントを使用して健康なボランティアで毛細血管リフィルのコースのより詳細な分析をお勧めします。

拡散反射率分光: 取得 1 つの親指の下の毛細血管リフィル テスト

The capillary refill test was introduced in 1947 to help estimate circulatory status in critically ill patients. Guidelines commonly state that refill ...

diffuse-reflectance-spectroscopy-getting-capillary-refill-test-under

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9.0K ビュー.  Region Östergötland. このプロトコルでは、拡散反射光の偏光分光学の使用が毛細血管リフィル テストの臨床的有用性を向上する方法について説明します。拡散反射率分光法ビデオと新しい情報学的エンドポイントを使用して健康なボランティアで毛細血管リフィルのコースのより詳細な分析をお勧めします。

ビデオ: Diffuse Reflectance Spectroscopy: Getting the Capillary Refill Test Under One's Thumb

Imaging Odor-Evoked Activities in the Mouse Olfactory Bulb using Optical Reflectance and Autofluorescence Signals

In the brain, sensory stimulation activates distributed populations of neurons among functional modules which participate to the coding of the stimulus. Functional optical imaging techniques are advantageous to visualize the activation of these modules in sensory cortices with high spatial resolution. In this context, endogenous optical signals that arise from molecular mechanisms linked to neuroenergetics are valuable sources of contrast to record spatial maps of sensory stimuli over wide fields in the rodent brain.
Here, we present two techniques based on changes of endogenous optical properties of the brain tissue during activation. First the intrinsic optical signals (IOS) are produced by a local alteration in red light reflectance due to: (i) absorption by changes in blood oxygenation level and blood volume (ii) photon scattering. The use of in vivo IOS to record spatial maps started in the mid 1980's with the observation of optical maps of whisker barrels in the rat and the orientation columns in the cat visual cortex1. IOS imaging of the surface of the rodent main olfactory bulb (OB) in response to odorants was later demonstrated by Larry Katz's group2. The second approach relies on flavoprotein autofluorescence signals (FAS) due to changes in the redox state of these mitochondrial metabolic intermediates. More precisely, the technique is based on the green fluorescence due to oxidized state of flavoproteins when the tissue is excited with blue light. Although such signals were probably among the first fluorescent molecules recorded for the study of brain activity by the pioneer studies of Britton Chances and colleagues3, it was not until recently that they have been used for mapping of brain activation in vivo. FAS imaging was first applied to the somatosensory cortex in rodents in response to hindpaw stimulation by Katsuei Shibuki's group4.
The olfactory system is of central importance for the survival of the vast majority of living species because it allows efficient detection and identification of chemical substances in the environment (food, predators). The OB is the first relay of olfactory information processing in the brain. It receives afferent projections from the olfactory primary sensory neurons that detect volatile odorant molecules. Each sensory neuron expresses only one type of odorant receptor and neurons carrying the same type of receptor send their nerve processes to the same well-defined microregions of &tilde;100&mu;m3 constituted of discrete neuropil, the olfactory glomerulus (Fig. 1). In the last decade, IOS imaging has fostered the functional exploration of the OB5, 6, 7 which has become one of the most studied sensory structures. The mapping of OB activity with FAS imaging has not been performed yet.
Here, we show the successive steps of an efficient protocol for IOS and FAS imaging to map odor-evoked activities in the mouse OB.

This article presents the protocols of intrinsic optical signals and flavoproteins autofluorescence signals imaging to map odor-evoked activities at the surface of the olfactory bulb in mice.

光反射率と自家蛍光信号を用いたマウス嗅球におけるイメージングの臭い誘発活動

In the brain, sensory stimulation activates distributed populations of neurons among functional modules which participate to the coding of the stimulus. ...

神経科学

The Mouse Forced Swim Test

The forced swim test is a rodent behavioral test used for evaluation of antidepressant drugs, antidepressant efficacy of new compounds, and experimental manipulations that are aimed at rendering or preventing depressive-like states. Mice are placed in an inescapable transparent tank that is filled with water and their escape related mobility behavior is measured. The forced swim test is straightforward to conduct reliably and it requires minimal specialized equipment. Successful implementation of the forced swim test requires adherence to certain procedural details and minimization of unwarranted stress to the mice. In the protocol description and the accompanying video, we explain how to conduct the mouse version of this test with emphasis on potential pitfalls that may be detrimental to interpretation of results and how to avoid them. Additionally, we explain how the behaviors manifested in the test are assessed.

The forced swim test is validated as an experimental approach to assess potential antidepressant efficacy in rodents. Experimental animals are placed in a tank of water and escape-related mobility behavior is quantified. The common procedures for the mouse version of this test are described.

マウス強制水泳試験

The forced swim test is a rodent behavioral test used for evaluation of antidepressant drugs, antidepressant efficacy of new compounds, and experimental ...

The Tail Suspension Test

The tail-suspension test is a mouse behavioral test useful in the screening of potential antidepressant drugs, and assessing of other manipulations that are expected to affect depression related behaviors. Mice are suspended by their tails with tape, in such a position that it cannot escape or hold on to nearby surfaces. During this test, typically six minutes in duration, the resulting escape oriented behaviors are quantified. The tail-suspension test is a valuable tool in drug discovery for high-throughput screening of prospective antidepressant compounds. Here, we describe the details required for implementation of this test with additional emphasis on potential problems that may occur and how to avoid them. We also offer a solution to the tail climbing behavior, a common problem that renders this test useless in some mouse strains, such as the widely used C57BL/6. Specifically, we prevent tail climbing behaviors by passing mouse tails through a small plastic cylinder prior to suspension. Finally, we detail how to manually score the behaviors that are manifested in this test.

The tail-suspension test is validated as an experimental procedure to assess antidepressant efficacy of drug treatments in mice. Mice are suspended by their tails for six minutes and escape-related behaviors are assessed. We describe procedures used in conducting the tail suspension test.

尾懸垂テスト

The tail-suspension test is a mouse behavioral test useful in the screening of potential antidepressant drugs, and assessing of other manipulations that ...

Laser Nanosurgery of Cerebellar Axons In Vivo

Only a few neuronal populations in the central nervous system (CNS) of adult mammals show local regrowth upon dissection of their axon. In order to understand the mechanism that promotes neuronal regeneration, an in-depth analysis of the neuronal types that can remodel after injury is needed. Several studies showed that damaged climbing fibers are capable of regrowing also in adult animals1,2. The investigation of the time-lapse dynamics of degeneration and regeneration of these axons within their complex environment can be performed by time-lapse two-photon fluorescence (TPF) imaging in vivo3,4. This technique is here combined with laser surgery, which proved to be a highly selective tool to disrupt fluorescent structures in the intact mouse cortex5-9.
This protocol describes how to perform TPF time-lapse imaging and laser nanosurgery of single axonal branches in the cerebellum in vivo. Olivocerebellar neurons are labeled by anterograde tracing with a dextran-conjugated dye and then monitored by TPF imaging through a cranial window. The terminal portion of their axons are then dissected by irradiation with a Ti:Sapphire laser at high power. The degeneration and potential regrowth of the damaged neuron are monitored by TPF in vivo imaging during the days following the injury.

Laser Nanosurgery of Cerebellar Axons In Vivo

Two-photon imaging, coupled to laser nanodissection, are useful tools to study degenerative and regenerative processes in the central nervous system with subcellular resolution. This protocol shows how to label, image, and dissect single climbing fibers in the cerebellar cortex in vivo.

小脳軸索のレーザーNanosurgery<em&gt;インビボ</em

Only a few neuronal populations in the central nervous system (CNS) of adult mammals show local regrowth upon dissection of their axon. In order to ...

The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

For most animals, feeding is an essential behavior for securing survival, and it influences development, locomotion, health and reproduction. Ingestion of the right type and quantity of food therefore has a major influence on quality of life. Research on feeding behavior focuses on the underlying processes that ensure actual feeding and unravels the role of factors regulating internal energy homeostasis and the neuronal bases of decision-making. The model organism Drosophila melanogaster, with its great variety of genetically traceable tools for labeling and manipulating single neurons, allows mapping of neuronal networks and identification of molecular signaling cascades involved in the regulation of food intake. This report demonstrates the CApillary FEeder assay (CAFE) and shows how to measure food intake in a group of flies for time spans ranging from hours to days. This easy-to-use assay consists of glass capillaries filled with liquid food that flies can freely access and feed on. Food consumption in the assay is accurately determined using simple measurement tools. Herein we describe step-by-step the method from setup to successful execution of the CAFE assay, and provide practical examples to analyze the food intake of a group of flies under controlled conditions. The reader is guided through possible limitations of the assay, and advantages and disadvantages of the method compared to other feeding assays in D. melanogaster are evaluated.

The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

The CApillary FEeder (CAFE) assay is a simple, budget-friendly, highly reliable method for investigating mechanisms underlying food intake. Used with the highly versatile genetic model organism Drosophila melanogaster, it provides a powerful means of gaining new insights into regulatory mechanisms of food intake.

キャピラリーフィーダーアッセイにおいて食物摂取を測定します<em&gt;キイロショウジョウバエ</em

For most animals, feeding is an essential behavior for securing survival, and it influences development, locomotion, health and reproduction. Ingestion of ...

Simultaneous Evaluation of Cerebral Hemodynamics and Light Scattering Properties of the In Vivo Rat Brain Using Multispectral Diffuse Reflectance Imaging

The simultaneous evaluation of cerebral hemodynamics and the light scattering properties of in vivo rat brain tissue is demonstrated using a conventional multispectral diffuse reflectance imaging system. This system is constructed from a broadband white light source, a motorized filter wheel with a set of narrowband interference filters, a light guide, a collecting lens, a video zoom lens, and a monochromatic charged-coupled device (CCD) camera. An ellipsoidal cranial window is made in the skull bone of a rat under isoflurane anesthesia to capture in vivo multispectral diffuse reflectance images of the cortical surface. Regulation of the fraction of inspired oxygen using a gas mixture device enables the induction of different respiratory states such as normoxia, hyperoxia, and anoxia. A Monte Carlo simulation-based multiple regression analysis for the measured multispectral diffuse reflectance images at nine wavelengths (500, 520, 540, 560, 570, 580, 600, 730, and 760 nm) is then performed to visualize the two-dimensional maps of hemodynamics and the light scattering properties of the in vivo rat brain.

Simultaneous Evaluation of Cerebral Hemodynamics and Light Scattering Properties of the In Vivo Rat Brain Using Multispectral Diffuse Reflectance Imaging

The simultaneous evaluation of cerebral hemodynamics and the light scattering properties of in vivo rat brain tissue is demonstrated using a conventional multispectral diffuse reflectance imaging system.

脳血行動態との光散乱特性の同時評価<em&gt;インビボ</emマルチスペクトル拡散反射率イメージングの使用&gt;ラット脳

The simultaneous evaluation of cerebral hemodynamics and the light scattering properties of in vivo rat brain tissue is demonstrated using a conventional ...

Functional Magnetic Resonance Spectroscopy at 7 T in the Rat Barrel Cortex During Whisker Activation

Nuclear magnetic resonance (NMR) spectroscopy offers the opportunity to measure cerebral metabolite contents in vivo and noninvasively. Thanks to technological developments over the last decade and the increase in magnetic field strength, it is now possible to obtain good resolution spectra in vivo in the rat brain. Neuroenergetics (i.e., the study of brain metabolism) and, especially, metabolic interactions between the different cell types have attracted more and more interest in recent years. Among these metabolic interactions, the existence of a lactate shuttle between neurons and astrocytes is still debated. It is, thus, of great interest to perform functional proton magnetic resonance spectroscopy (1H-MRS) in a rat model of brain activation and monitor lactate. However, the methyl lactate peak overlaps lipid resonance peaks and is difficult to quantify. The protocol described below allows metabolic and lactate fluctuations to be monitored in an activated brain area. Cerebral activation is obtained by whisker stimulation and 1H-MRS is performed in the corresponding activated barrel cortex, whose area is detected using blood-oxygen-level-dependent functional magnetic resonance imaging (BOLD fMRI). All steps are fully described: the choice of anesthetics, coils, and sequences, achieving efficient whisker stimulation directly in the magnet, and data processing.

After checking by blood-oxygen-level-dependent functional magnetic resonance imaging (BOLD fMRI) that the corresponding somatosensory barrel field cortex area (called S1BF) is correctly activated, the main goal of this study is to quantify lactate content fluctuations in the activated rat brains by localized proton magnetic resonance spectroscopy (1H-MRS) at 7 T.

7 機能的磁気共鳴分光ウィスカ アクティベーション中にラットのバレル皮質の T

Nuclear magnetic resonance (NMR) spectroscopy offers the opportunity to measure cerebral metabolite contents in vivo and noninvasively. Thanks to ...

Ex Vivo Pressurized Hippocampal Capillary-Parenchymal Arteriole Preparation for Functional Study

From subtle behavioral alterations to late-stage dementia, vascular cognitive impairment typically develops following cerebral ischemia. Stroke and cardiac arrest are remarkably sexually dimorphic diseases, and both induce cerebral ischemia. However, progress in understanding the vascular cognitive impairment, and then developing sex-specific treatments, has been partly limited by challenges in investigating the brain microcirculation from mouse models in functional studies. Here, we present an approach to examine the capillary-to-arteriole signaling in an ex vivo hippocampal capillary-parenchymal arteriole (HiCaPA) preparation from mouse brain. We describe how to isolate, cannulate, and pressurize the microcirculation to measure arteriolar diameter in response to capillary stimulation. We show which appropriate functional controls can be used to validate the HiCaPA preparation integrity and display typical results, including testing potassium as a neurovascular coupling agent and the effect of the recently characterized inhibitor of the Kir2 inward rectifying potassium channel family, ML133. Further, we compare the responses in preparations obtained from male and female mice. While these data reflect functional investigations, our approach can also be used in molecular biology, immunochemistry, and electrophysiology studies.

The present manuscript details how to isolate hippocampal arterioles and capillaries from the mouse brain and how to pressurize them for pressure myography, immunofluorescence, biochemistry, and molecular studies.

機能的研究のためのEx Vivo加圧海馬毛細血管-パンレンキマル動脈製剤

From subtle behavioral alterations to late-stage dementia, vascular cognitive impairment typically develops following cerebral ischemia. Stroke and ...

Induction of Diffuse Axonal Brain Injury in Rats Based on Rotational Acceleration

Traumatic brain injury (TBI) is a major cause of death and disability. Diffuse axonal injury (DAI) is the predominant mechanism of injury in a large percentage of TBI patients requiring hospitalization. DAI involves widespread axonal damage from shaking, rotation or blast injury, leading to rapid axonal stretch injury and secondary axonal changes that are associated with a long-lasting impact on functional recovery. Historically, experimental models of DAI without focal injury have been difficult to design. Here we validate a simple, reproducible and reliable rodent model of DAI that causes widespread white matter damage without skull fractures or contusions.

This protocol validates a reliable, easy-to-perform and reproducible rodent model of brain diffuse axonal injury (DAI) that induces widespread white matter damage without skull fractures or contusions.

回転加速度に基づくラットにおけるびまん軸索脳損傷の誘導

Traumatic brain injury (TBI) is a major cause of death and disability. Diffuse axonal injury (DAI) is the predominant mechanism of injury in a large ...

Culture of Brain Capillary Pericytes for Cytosolic Calcium Measurements and Calcium Imaging Studies

Pericytes are associated with endothelial cells and astrocytic endfeet in a structure known as the neurovascular unit (NVU). Brain capillary pericyte function is not fully known. Pericytes have been suggested to be involved in capillary development, regulation of endothelial barrier tightness and trancytosis activity, regulation of capillary tone and to play crucial roles in certain brain pathologies.
Pericytes are challenging to investigate in the intact brain due to the difficulties in visualizing processes in the brain parenchyma, as well as the close proximity to the other cells of the NVU. The present protocol describes a method for isolation and culture of primary bovine brain capillary pericytes and their following usage in calcium imaging studies, where effects of agonists involved in brain signaling and pathologies can be investigated. Cortical capillary fragments are allowed to attach to the bottom of culture flasks and, after 6 days, endothelial cells and pericytes have grown out from the capillary fragments. The endothelial cells are removed by gentle trypsinization and pericytes are cultured for 5 additional days before passaging.
Isolated pericytes are seeded in 96-well culture plates and loaded with the calcium indicator dye (Fura-2 acetoxymethyl (AM)) to allow for measurements of intracellular calcium levels in a plate reader setup. Alternatively, pericytes are seeded on coverslips and mounted in cell chambers. Following loading with the calcium indicator (Cal-520 AM), calcium live-imaging can be performed using confocal microscopy at an excitation wavelength of 488 nm and emission wavelength of 510-520 nm.
The method described here has been used to obtain the first intracellular calcium measurements from primary brain capillary pericytes, demonstrating that pericytes are stimulated via ATP and are able to contract in vitro.

Brain capillary pericytes are essential players in the regulation of blood-brain barrier properties and blood flow. This protocol describes how brain capillary pericytes can be isolated, cultured, characterized with respect to cell type and applied for investigations of intracellular calcium signaling with fluorescent probes.

細胞体カルシウム測定とカルシウムイメージング研究のための脳毛細血管培養

Pericytes are associated with endothelial cells and astrocytic endfeet in a structure known as the neurovascular unit (NVU). Brain capillary pericyte ...

拡散反射率分光: 取得 1 つの親指の下の毛細血管リフィル テスト

拡散反射率分光: 取得 1 つの親指の下の毛細血管リフィルテスト