We wish to thank the study participants for making this work possible and the staff at the NIHR Southampton Biomedical Research Centre and the Wellcome Trust Clinical Research Facility, University Hospital Southampton for assistance with study measurements.
This work was supported by the NIHR Southampton Biomedical Research Centre, Nutrition, the MRC Lifecourse Epidemiology Unit and the University of Southampton. This report is independent research by the NIHR BRC Funding Scheme. The views expressed in this publication are those of the authors and not necessarily those of the NHS, the NIHR or the Department of Health. AMB is supported by a NIHR Clinical Research Fellowship, HPP is supported by the NIHR Southampton Biomedical Research Centre.

倫理の声明：我々は我々の施設内に従う筋生検手順の説明は、現在次の。筋生検手順は、ハートフォードシャー州調査委員会番号07 / Q0204 / 68によって承認されたハートフォードシャー州サルコペニア研究で使用されています。すべての参加者は書面によるインフォームドコンセント20を与えました。 注：単一のオペレータは、不安を軽減し、彼らはすべての回で快適であることを確実にするために、会話の中で参加者を従事することができアシスタントによって助け手順を実行することができます。処置室は、シンク、高さ調節可能なソファ、清浄な表面と手順トロリーに装備する必要があります。 1.参加者を準備します（図2） <ol><li>ソファの上に仰向け、快適に配置するために参加者を確認して下さい。鼠径部の折り目から優先太ももを公開します。露出した太ももの下に使い捨て吸収性シートを広げます。脚は手順を通してまっすぐでリラックスしたままであることを確認しますが、太ももは、 外側広筋の概要を説明するために一時的に緊張されています。 </li><li>マーク約2/3膝蓋骨に前腸骨棘から仮想線ダウン手順サイト。 </li></ol> 2.スキンを準備し（図2） <ol><li>無菌のシェービング刃で提案された生検部位を囲む直径約4 cmの皮膚を剃るとアルコール綿で皮膚をきれいに。 注：これは、治癒過程中の創傷における後続の毛捕捉を防止するためです。このステップは、操作者の好みに応じて任意です。 </li><li>局所麻酔薬で皮膚と上に筋膜を潜入。その後、23 G針で皮下組織に深く浸透し、2％のリドカインの皮下ブレブを上げるために25 G針を使用してください。生検部位での皮下組織含有量に応じて1インチに少なくとも半分に浸透することを目指しています。この深さは、上にFASを確実にするために十分であるべきですCIAは麻酔されています。アクションへの麻酔薬のために5分 - 2を許可します。針または手術用メスの刃の先端で軽く肌をプロービングすることによって麻酔を確認してください。 </li></ol> 3.滅菌野を準備します（図2） <ol><li>石鹸で手を洗い、滅菌ガウンを着用されるまで、清潔な手を維持します。手順トロリーに滅菌フィールドを作成する前に滅菌手袋と手をカバーしています。 </li><li> 2％クロルヘキシジン/イソプロピルアルコール溶液またはヨウ素ベースの皮膚の消毒剤と生検部位を滅菌します。生検部位を露出させるだけでなく、無菌領域を維持するために、接着剤開口部と滅菌ドレープを適用します。 </li></ol> 4.手順を実行します（図2） <ol><li>ダウンとその上の筋膜を介して、皮膚上の10ミリメートルの切開 - サイズ11メスで、5を作ります。深さ平均の半分に1インチに、大腿骨の長軸に対して直角に切開を通してconchotomeの閉じた痛烈な先端を挿入します。必要な使用している場合conchotomeの先端は、筋肉内に開閉される一方で、生検部位の周囲の大腿部を保持し、サポートするために、フリーハンド。 </li><li>筋肉をカットして180° - 90を介してconchotomeを回転させます。 conchotomeを撤回し、無菌生理食塩水によって減衰滅菌ガーゼ上に先端を開きます。必要に応じて十分な筋肉組織を得るために、単一の創傷部位内の手順を繰り返します。氷の上に置かれた容器の中にガーゼを転送します。地元の機関のプロトコルに従って筋肉を処理するための準備室にコンテナを転送します。 </li></ol> 5.ポスト​​手順（図2） <ol><li>最大5分間創傷に直接圧力を適用します。直角とは対照的にsteriストリップが誤って削除された場合、潜在的に瘢痕組織を除去し、傷を再び開く回避するために、創傷に平行に配置してsteri-ストリップで傷口を閉じます。 steriストリップに滅菌吸収性包帯を置き、二層を結びます圧縮のために包帯やテープで固定します。 </li><li> 4日手順の後 - 彼らは3ドレッシングの変更を実行する必要がありますよう参加者に創傷ドレッシング方法を説明します。参加者は後でその夜寝る前に圧迫包帯を削除していることを確認してください。 </li></ol> 6.ポスト生検を見ます<ol><li>穏やかな運動（ 例えば 、歩行）によって緩和することができるいくつかの太ももの剛性を体験することが一般的であることを参加者に通知します。アセトアミノフェンなどの単純な鎮痛を使用してください。 2週間まで持続し得る生検部位の周りの過渡しびれだけでなく、創傷感染の潜在的な合併症についての参加者を警告します。 </li></ol>件名POST手順7.手順<ol><li> （このような登山、ランニング、重い物を持ち上げるように）72時間精力的な活動を避けるために、参加者を確認して下さい。 72時間 - 48水に浸漬を避けるために、参加者を確認して下さい。しかし、participanを指示ドレッシングの乾燥を保つためにシャワーを有するときに生検部位の周りにラップフィルムまたは類似をラップするトン。 </li><li> 4日と1週間後steriストリップを削除 - 3の後にドレッシングを変更するには、参加者を確認して下さい。 注：参加者は、ポスト生検ケアのための説明書、予備のドレッシングや手順を実行を担当するチームの緊急連絡先の詳細を与えられるべきです。 </li></ol>

<ol>
	<li>Dubowitz, V., Sewry, C. A., Oldfords, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Muscle Biopsy: A Practical Approach. , (2013).</li><li>Parent, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Duchenne+De+Boulogne:+a+pioneer+in+neurology+and+medical+photography.">Duchenne De Boulogne: a pioneer in neurology and medical photography.</a> Can J Neurol Sci. 32 (3), 369-377 (2005).</li><li>Hayot, M., Michaud, A., Koechlin, C., Caron, M. A., Leblanc, P., Prefaut, C., Maltais, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Skeletal+muscle+microbiopsy:+a+validation+study+of+a+minimally+invasive+technique.">Skeletal muscle microbiopsy: a validation study of a minimally invasive technique.</a> Eur Respir J. 25 (3), 431-440 (2005).</li><li>O'Rourke, K. S., Blaivas, M., Ike, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Utility+of+needle+muscle+biopsy+in+a+university+rheumatology+practice.">Utility of needle muscle biopsy in a university rheumatology practice.</a> J Rheumatol. 21 (3), 413-424 (1994).</li><li>O'Rourke, K. S., Ike, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscle+biopsy.">Muscle biopsy.</a> Curr Opin Rheumatol. 7 (6), 462-468 (1995).</li><li>Bergstrom, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscle+electrolytes+in+man.">Muscle electrolytes in man.</a> Scand J Clin Lab Invest. 14, (1962).</li><li>Edwards, R., Young, A., Wiles, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Needle+biopsy+of+skeletal+muscle+in+the+diagnosis+of+myopathy+and+the+clinical+study+of+muscle+function+and+repair.">Needle biopsy of skeletal muscle in the diagnosis of myopathy and the clinical study of muscle function and repair.</a> N Engl J Med. 302 (5), 261-271 (1980).</li><li>Edwards, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Percutaneous+needle-biopsy+of+skeletal+muscle+in+diagnosis+and+research.">Percutaneous needle-biopsy of skeletal muscle in diagnosis and research.</a> Lancet. 2 (7724), 593-595 (1971).</li><li>Edwards, R. H., Round, J. M., Jones, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Needle+biopsy+of+skeletal+muscle:+a+review+of+10+years+experience.">Needle biopsy of skeletal muscle: a review of 10 years experience.</a> Muscle Nerve. 6 (9), 676-683 (1983).</li><li>Shanely, R. A., Zwetsloot, K. A., Triplett, N. T., Meaney, M. P., Farris, G. E., Nieman, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+skeletal+muscle+biopsy+procedures+using+the+modified+Bergstrom+technique.">Human skeletal muscle biopsy procedures using the modified Bergstrom technique.</a> J Vis Exp. (91), e51812 (2014).</li><li>Coggan, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscle+biopsy+as+a+tool+in+the+study+of+aging.">Muscle biopsy as a tool in the study of aging.</a> J Gerontol A Biol Sci Med Sci. 50, 30-34 (1995).</li><li>Henriksson, K. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term='Semi-open'+muscle+biopsy+technique.+A+simple+outpatient+procedure.">'Semi-open' muscle biopsy technique. A simple outpatient procedure.</a> Acta Neurol Scand. 59 (6), 317-323 (1979).</li><li>Dorph, C., Nennesmo, I., Lundberg, I. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Percutaneous+conchotome+muscle+biopsy.+A+useful+diagnostic+and+assessment+tool.">Percutaneous conchotome muscle biopsy. A useful diagnostic and assessment tool.</a> J Rheumatol. 28 (7), 1591-1599 (2001).</li><li>Poulsen, M. B., Bojsen-Moller, M., Jakobsen, J., Andersen, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Percutaneous+conchotome+biopsy+of+the+deltoid+and+quadricep+muscles+in+the+diagnosis+of+neuromuscular+disorders.">Percutaneous conchotome biopsy of the deltoid and quadricep muscles in the diagnosis of neuromuscular disorders.</a> J Clin Neuromuscul Dis. 7 (1), 36-41 (2005).</li><li>Dietrichson, P., Mellgren, S. I., Skre, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscle+biopsy+with+the+percutaneous+conchotome+technique.">Muscle biopsy with the percutaneous conchotome technique.</a> J Oslo City Hosp. 30 (5), 73-79 (1980).</li><li>Dietrichson, P., Coakley, J., Smith, P. E., Griffiths, R. D., Helliwell, T. R., Edwards, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conchotome+and+needle+percutaneous+biopsy+of+skeletal+muscle.">Conchotome and needle percutaneous biopsy of skeletal muscle.</a> J Neurol Neurosurg Psychiatry. 50 (11), 1461-1467 (1987).</li><li>Andonopoulos, A. P., Papadimitriou, C., Melachrinou, M., Meimaris, N., Vlahanastasi, C., Bounas, A., Georgiou, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Asymptomatic+gastrocnemius+muscle+biopsy:+an+extremely+sensitive+and+specific+test+in+the+pathologic+confirmation+of+sarcoidosis+presenting+with+hilar+adenopathy.">Asymptomatic gastrocnemius muscle biopsy: an extremely sensitive and specific test in the pathologic confirmation of sarcoidosis presenting with hilar adenopathy.</a> Clin Exp Rheumatol. 19 (5), 569-572 (2001).</li><li>Helliwell, T. R., Coakley, J., Smith, P. E., Edwards, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+morphology+and+morphometry+of+the+normal+human+tibialis+anterior+muscle.">The morphology and morphometry of the normal human tibialis anterior muscle.</a> Neuropathol Appl Neurobiol. 13 (4), 297-307 (1987).</li><li>Patel, H. P., Jameson, K. A., Syddall, H. E., Martin, H. J., Stewart, C. E., Cooper, C., Sayer, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developmental+influences,+muscle+morphology,+and+sarcopenia+in+community-dwelling+older+men.">Developmental influences, muscle morphology, and sarcopenia in community-dwelling older men.</a> J Gerontol A Biol Sci Med Sci. 67 (1), 82-87 (2012).</li><li>Patel, H. P., Syddall, H. E., Martin, H. J., Stewart, C. E., Cooper, C., Sayer, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hertfordshire+Sarcopenia+Study:+design+and+methods.">Hertfordshire Sarcopenia Study: design and methods.</a> BMC Geriatrics. 10 (1), 43 (2010).</li><li>Anderson, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recommendations+for+the+biopsy+procedure+and+assessment+of+skeletal+muscle+biopsies.">Recommendations for the biopsy procedure and assessment of skeletal muscle biopsies.</a> Virchows Arch. 431 (4), 227-233 (1997).</li><li>Patel, H. P., Syddall, H. E., Martin, H. J., Cooper, C., Stewart, C., Sayer, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Feasibility+and+Acceptability+of+Muscle+Biopsy+in+Epidemiological+Studies:+Findings+from+the+Hertfordshire+Sarcopenia+Study+(HSS).">The Feasibility and Acceptability of Muscle Biopsy in Epidemiological Studies: Findings from the Hertfordshire Sarcopenia Study (HSS).</a> J Nutr Health Aging. 15 (1), 10-15 (2011).</li><li>Charifi, N., Kadi, F., Feasson, L., Costes, F., Geyssant, A., Denis, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhancement+of+microvessel+tortuosity+in+the+vastus+lateralis+muscle+of+old+men+in+response+to+endurance+training.">Enhancement of microvessel tortuosity in the vastus lateralis muscle of old men in response to endurance training.</a> J Physiol. 554 (Pt 2), 559-569 (2004).</li><li>Frontera, W. R., Suh, D., Krivickas, L. S., Hughes, V. A., Goldstein, R., Roubenoff, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Skeletal+muscle+fiber+quality+in+older+men+and+women.">Skeletal muscle fiber quality in older men and women.</a> Am J Physiol Cell Physiol. 279 (3), C611-C618 (2000).</li><li>He, J., Watkins, S., Kelley, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Skeletal+muscle+lipid+content+and+oxidative+enzyme+activity+in+relation+to+muscle+fiber+type+in+type+2+diabetes+and+obesity.">Skeletal muscle lipid content and oxidative enzyme activity in relation to muscle fiber type in type 2 diabetes and obesity.</a> Diabetes. 50 (4), 817-823 (2001).</li><li>Patel, H. P., Al-Shanti, N., Davies, L. C., Barton, S. J., Grounds, M. D., Tellam, R. L., Stewart, C. E., Cooper, C., Sayer, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lean+mass,+muscle+strength+and+gene+expression+in+community+dwelling+older+men:+findings+from+the+Hertfordshire+Sarcopenia+Study+(HSS).">Lean mass, muscle strength and gene expression in community dwelling older men: findings from the Hertfordshire Sarcopenia Study (HSS).</a> Calcif Tissue Int. 95 (4), 308-316 (2014).</li><li>Peterson, C. M., Johannsen, D. L., Ravussin, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Skeletal+muscle+mitochondria+and+aging:+a+review.">Skeletal muscle mitochondria and aging: a review.</a> J Aging Res. 2012, 194821 (2012).</li></ol>

None declared.

参加者の安全を確保し、効率的に筋生検を行うためには、プロトコル内の重要なステップに注意を払うことが重要です。 参加者は慎重に、このような出血や創傷治癒不良などの合併症を避けるために除外基準を考慮した後に選択する必要があります。手順全体で厳格な無菌状態が不可欠です。正しい技術の適用は、参加者の快適性を確保参加者の組織に不要な外傷を最?...

ミオパシーおよび他の神経筋変性疾患の診断のための骨格筋を取得することは許容され、患者1に無能力化されない安全で効率的な方法に依存しています。歴史的に、分析のための筋肉組織を得る方法は、全身麻酔下または死後から実行オープン生検が含まれていました。これらの技術は、筋肉の直接可視化を可能にし、筋肉の大きい作品は生検することが許可しました。これらの技術は入院と回復の点で患者に持っている潜在的な欠点を考えると、経皮的な「セミオープン」筋生検法は、代替として導入されました。この技術は、最初にトロカール自己構築針を使用し、全身麻酔2せずに生体からの筋肉サンプルを得ることができたデュシェンヌ（1806から1875）により行いました。その後、生検針の様々な形態は、3,4,5使用されています。しかし、生検nは1962年にバーグストロムによって開発eedleは、最も一般的に臨床で使用されています。両方の子供と大人でだけでなく、研究6,7,8,9,10インチこれは、組織のポスト生検を追放するために鋭利なトロカール、切断カニューレとプッシュロッドを有しています。この手順から得られた筋肉の収率は繰り返しサンプリング11,9後25 -293ミリグラムの範囲であると報告されています。 Henrikssonは、代替セミオープン筋生検法（ 図1）12として1979年にワイル・ブレークスリーのconchotomeを導入しました。機器はバーグストロムとは異なり、単一の構造の構築であり、シャープな痛烈な先端を有する鉗子のように設計されています。チップサイズを4mmの幅が6mmの範囲であることができます。痛烈な先端の2つの縁部が反対した場合、中空は筋肉のキャプチャを確実に形成されています。 conchotome 5を通って挿入される - 10mMの皮膚切開とscalpeなどの筋肉とその上の筋膜を貫通するためのトロカールのための必要性を回避しますLは、筋肉への追跡を行うために使用することができます。 この方法は、操作性13,14度の高い組織浸透を制御することができます。これは、過度の圧力が潜在的に神経血管または基礎骨構造などを破損する恐れがあります。、前脛骨15,14の部位でのサイトのために特に有用です。 conchotomeを使用して、筋肉の収率が可変することができ、私たちの実際に20 - 筋の200 mgを繰り返しサンプリングした後に得られています。ベルイストロームの針の上conchotomeの1つの潜在的な利点は、それが提供する機動性の高い程度に加えて、痛烈な先端が先鋭化やバーグストロムのトロカール16を構築ほど頻繁に交換する必要がないことです。 ワイルブレークスリーのconchotomeを使用して、筋生検の全体的な目標は、診断およびresearの両方のための組織化学および分子分析を可能にするために筋肉組織の十分な量を得ることですchの研究。これは、簡単に学ぶことができる簡単で安全な手技です。この技術では、生検に適した筋肉は上腕二頭筋、三頭筋、三角筋、腓腹筋、前脛骨筋、ヒラメ筋とsacrospinals 17,15,7,18,14が含まれています。それは太もも内の主な神経血管構造を回避するように外側広筋の最も外側の部分は、生検のために使用される最も一般的なサイトです。これは、上前腸骨棘と膝蓋骨を結合する仮想線の下の約3分の2に識別されます。筋肉形状データは、 広筋由来例myofibreエリア、myofibre比率、毛細血管密度のためのサンプリングは研究11,19間の比較を可能にする文献で ​​広く入手可能である外側の 。

<table><tbody><tr><td>Weil-Blakesley conchotome</td><td>Gebr&amp;uuml;der Zepf Medizintechnik, D&amp;uuml;rbheim, Germany</td><td>None</td><td>6 mm biting tip</td></tr><tr><td>5 ml 2% lidocaine</td><td>Generic use</td><td>None</td><td>One 5 ml vial should suffice for adequate anaesthesia</td></tr><tr><td>Alcohol swab</td><td>Generic use</td><td>None</td><td></td></tr><tr><td>5 ml syringe</td><td>Generic use</td><td>None</td><td></td></tr><tr><td>25 G and 23 G sterile needles</td><td>Generic use</td><td>None</td><td></td></tr><tr><td>Sterile scalpel size 11</td><td>Generic use</td><td>None</td><td></td></tr><tr><td>Sterile gauze squares</td><td>Generic use</td><td>None</td><td></td></tr><tr><td>Sodium chloride 0.9% 10 ml</td><td>Fannin</td><td>1122/002/001</td><td></td></tr><tr><td>Steri-Drape&amp;trade; Small Drape with Adhesive Aperture</td><td>3M</td><td>1020</td><td></td></tr><tr><td>ChloraPrep 2% w/v /70% v/v cutaneous solution&amp;nbsp;</td><td>CareFusion</td><td>270400</td><td>Two</td></tr><tr><td>Steri-Strip&amp;trade; 1/4 inch x 3 inch Reinforced Adhesive Skin Closures</td><td>3M</td><td>R1541</td><td></td></tr><tr><td>PRIMAPORE sterile dressing 15 cm x 8 cm</td><td>Smith &amp;amp;&amp;nbsp;&lt;br/&gt; Nephew</td><td>66000318</td><td></td></tr><tr><td>Cotton crepe bandage Hospicrepe 233 7,5 cm x 4,5 m</td><td>Hartmann</td><td>915562</td><td></td></tr><tr><td>Flexible cohesive bandage Co-Plus LF</td><td>BSN medical</td><td>2904565</td><td></td></tr><tr><td>Disposable absorbent sheet</td><td>Generic use</td><td>None</td><td></td></tr><tr><td>Micropore or transpore tape</td><td>Generic use</td><td>None</td><td></td></tr><tr><td>Sterile gown with sterile paper towels</td><td>Generic use</td><td>None</td><td></td></tr><tr><td>Sterile gloves</td><td>Generic use</td><td>None</td><td></td></tr></tbody></table>

human vastus lateralis skeletal muscle biopsy using the weil-blakesley conchotome

ワイル・ブレークスリーのconchotomeを使用して、経皮的筋生検はよく両方の臨床と研究の実践において確立されています。それは、安全で効果的かつ良好な忍容手法です。 6ミリメートル幅の広い中空 - ワイル・ブレークスリーのconchotomeは4シャープな痛烈な先端を有します。これは、5を通って挿入される - 10mMの皮膚切開と制御組織浸透のために操作することができます。先端が開閉組織内に、次に筋肉を切断する90 -180°回転されます。繰り返しサンプリング後に得られた筋肉の量は、20mgのから組織学および分子生物学的研究の両方のために処理することができる290 mgまで変化させることができます。傷は活動の正常レベルは、手順の直後に再起動することができますが、約72時間の最小に抑え、乾燥し、活発な身体活動を保つ必要があります。細心の注意を被験者、完全な無菌状態とポスト手続きのケアの選択に支払わされている場合、この手順は、安全かつ効果的です。左右両方Vastusの外側のは、参加者の好みに応じて生検に適しています。

上記の手順は、両方の臨床と研究の設定において安全かつ許容可能です。生検が診断プロセスの一部として実行されると、筋肉が患者の症状や筋力低下の兆候に応じて選択すべきです。生検部位は、前の傷害、拘縮または計測21,1から自由でなければなりません。研究では、糖尿病患者の空腹時又は除外のような標準化された条件は、19を必要とするこ...

Watch this Scientific Journal Video about Human Vastus Lateralis Skeletal Muscle Biopsy Using the Weil-Blakesley Conchotome at JoVE.com

Human Vastus Lateralis Skeletal Muscle Biopsy Using the Weil-Blakesley Conchotome

This video demonstrates the technique of percutaneous muscle biopsy of the human vastus lateralis using the Weil-Blakesley conchotome.

人間<em&gt;外側広筋</emワイル・ブレークスリーConchotomeを使用&gt;骨格筋生検

Percutaneous muscle biopsy using the Weil-Blakesley conchotome is well established in both clinical and research practice. It is a safe, effective and ...

human-vastus-lateralis-skeletal-muscle-biopsy-using-weil-blakesley

Research

JoVE Journal

Medicine

Human Vastus Lateralis Skeletal Muscle Biopsy Using the Weil-Blakesley Conchotome

16.6K ビュー.  University of Southampton, University Hospital Southampton. This video demonstrates the technique of percutaneous muscle biopsy of the human vastus lateralis using the Weil-Blakesley conchotome.

ビデオ: Human Vastus Lateralis Skeletal Muscle Biopsy Using the Weil-Blakesley Conchotome

Isolation and Quantitative Immunocytochemical Characterization of Primary Myogenic Cells and Fibroblasts from Human Skeletal Muscle

The repair and regeneration of skeletal muscle requires the action of satellite cells, which are the resident muscle stem cells. These can be isolated from human muscle biopsy samples using enzymatic digestion and their myogenic properties studied in culture. Quantitatively, the two main adherent cell types obtained from enzymatic digestion are: (i) the satellite cells (termed myogenic cells or muscle precursor cells), identified initially as CD56+ and later as CD56+/desmin+ cells and (ii) muscle-derived fibroblasts, identified as CD56&#8211; and TE-7+. Fibroblasts proliferate very efficiently in culture and in mixed cell populations these cells may overrun myogenic cells to dominate the culture. The isolation and purification of different cell types from human muscle is thus an important methodological consideration when trying to investigate the innate behavior of either cell type in culture. Here we describe a system of sorting based on the gentle enzymatic digestion of cells using collagenase and dispase followed by magnetic activated cell sorting (MACS) which gives both a high purity (&#62;95% myogenic cells) and good yield (~2.8 x 106 &#177; 8.87 x 105 cells/g tissue after 7 days in vitro) for experiments in culture. This approach is based on incubating the mixed muscle-derived cell population with magnetic microbeads beads conjugated to an antibody against CD56 and then passing cells though a magnetic field. CD56+ cells bound to microbeads are retained by the field whereas CD56&#8211; cells pass unimpeded through the column. Cell suspensions from any stage of the sorting process can be plated and cultured. Following a given intervention, cell morphology, and the expression and localization of proteins including nuclear transcription factors can be quantified using immunofluorescent labeling with specific antibodies and an image processing and analysis package.

The main adherent cell types derived from human muscle are myogenic cells and fibroblasts. Here, cell populations are enriched using magnetic-activated cell sorting based on the CD56 antigen. Subsequent immunolabelling with specific antibodies and use of image analysis techniques allows quantification of cytoplasmic and nuclear characteristics in individual cells.

ヒト骨格筋からの一次筋原細胞および線維芽細胞の単離および定量的免疫組織化学的特性評価

The repair and regeneration of skeletal muscle requires the action of satellite cells, which are the resident muscle stem cells. These can be isolated ...

発生生物学

Tibial Nerve Transection - A Standardized Model for Denervation-induced Skeletal Muscle Atrophy in Mice

The tibial nerve transection model is a well-tolerated, validated, and reproducible model of denervation-induced skeletal muscle atrophy in rodents. Although originally developed and used extensively in the rat due to its larger size, the tibial nerve in mice is big enough that it can be easily manipulated with either crush or transection, leaving the peroneal and sural nerve branches of the sciatic nerve intact and thereby preserving their target muscles. Thus, this model offers the advantages of inducing less morbidity and impediment of ambulation than the sciatic nerve transection model and also allows investigators to study the physiologic, cellular and molecular biologic mechanisms regulating the process of muscle atrophy in genetically engineered mice. The tibial nerve supplies the gastrocnemius, soleus and plantaris muscles, so its transection permits the study of denervated skeletal muscle composed of fast twitch type II fibers and/or slow twitch type I fibers. Here we demonstrate the tibial nerve transection model in the C57Black6 mouse. We assess the atrophy of the gastrocnemius muscle, as a representative muscle, at 1, 2, and 4 weeks post-denervation by measuring muscle weights and fiber type specific cross-sectional area on paraffin-embedded histologic sections immunostained for fast twitch myosin.

The tibial nerve transection model is a well-tolerated, validated, and reproducible model of skeletal muscle atrophy. The model surgical protocol is described and demonstrated in C57Black6 mice. &#160;

脛骨神経切断 - マウスにおける脱神経誘発性骨格筋萎縮のために標準化されたモデル

The tibial nerve transection model is a well-tolerated, validated, and reproducible model of denervation-induced skeletal muscle atrophy in rodents. ...

医学

Isolation, Culture, Characterization, and Differentiation of Human Muscle Progenitor Cells from the Skeletal Muscle Biopsy Procedure

The use of primary human tissue and cells is ideal for the investigation of biological and physiological processes such as the skeletal muscle regenerative process. There are recognized challenges to working with human primary adult stem cells, particularly human muscle progenitor cells (hMPCs) derived from skeletal muscle biopsies, including low cell yield from collected tissue and a large degree of donor heterogeneity of growth and death parameters among cultures. While incorporating heterogeneity into experimental design requires a larger sample size to detect significant effects, it also allows us to identify mechanisms that underlie variability in hMPC&#160;expansion capacity, and thus allows us to better understand heterogeneity in skeletal muscle regeneration. Novel mechanisms that distinguish the expansion capacity of cultures have the potential to lead to the development of therapies to improve skeletal muscle regeneration.

We present techniques for isolating, culturing, characterizing, and differentiating human primary muscle progenitor cells (hMPCs) obtained from skeletal muscle biopsy tissue. hMPCs obtained and characterized through these methods can be used to subsequently address research questions related to human myogenesis and skeletal muscle regeneration.

骨格筋生検手順からのヒト筋前駆細胞の単離、培養、特徴付け、分化

The use of primary human tissue and cells is ideal for the investigation of biological and physiological processes such as the skeletal muscle ...

Human Skeletal Muscle Biopsy Procedures Using the Modified Bergstr&#246;m Technique

The percutaneous biopsy technique enables researchers and clinicians to collect skeletal muscle tissue samples. The technique is safe and highly effective. This video describes the percutaneous biopsy technique using a modified Bergstr&#246;m needle to obtain skeletal muscle tissue samples from the vastus lateralis of human subjects. The Bergstr&#246;m needle consists of an outer cannula with a small opening (&#8216;window&#8217;) at the side of the tip and an inner trocar with a cutting blade at the distal end. Under local anesthesia and aseptic conditions, the needle is advanced into the skeletal muscle through an incision in the skin, subcutaneous tissue, and fascia. Next, suction is applied to the inner trocar, the outer trocar is pulled back, skeletal muscle tissue is drawn into the window of the outer cannula by the suction, and the inner trocar is rapidly closed, thus cutting or clipping the skeletal muscle tissue sample. The needle is rotated 90&#176;&#160;and another cut is made. This process may be repeated three more times. This multiple cutting technique typically produces a sample of 100-200 mg or more in healthy subjects and can be done immediately before, during, and after a bout of exercise or other intervention. Following post-biopsy dressing of the incision site, subjects typically resume their activities of daily living right away and can fully participate in vigorous physical activity within 48-72 hr. Subjects should avoid heavy resistance exercise for 48 hr to reduce the risk of herniation of the muscle through the incision in the fascia.

The purpose of this video is to present the modified Bergstr&#246;m skeletal muscle biopsy technique on human subjects.

変更されたベルイストローム法を用いたヒト骨格筋生検手順

The percutaneous biopsy technique enables researchers and clinicians to collect skeletal muscle tissue samples. The technique is safe and highly ...

Preparation and Respirometric Assessment of Mitochondria Isolated from Skeletal Muscle Tissue Obtained by Percutaneous Needle Biopsy

Respirometric profiling of isolated mitochondria is commonly used to investigate electron transport chain function. We describe a method for obtaining samples of human Vastus lateralis, isolating mitochondria from minimal amounts of skeletal muscle tissue, and plate based respirometric profiling using an extracellular flux (XF) analyzer. Comparison of respirometric profiles obtained using 1.0, 2.5 and 5.0 &#956;g of mitochondria indicate that 1.0 &#956;g is sufficient to measure respiration and that 5.0 &#956;g provides most consistent results based on comparison of standard errors. Western blot analysis of isolated mitochondria for mitochondrial marker COX IV and non-mitochondrial tissue marker GAPDH indicate that there is limited non-mitochondrial contamination using this protocol. The ability to study mitochondrial respirometry in as little as 20 mg of muscle tissue allows users to utilize individual biopsies for multiple study endpoints in clinical research projects.

Methods for biopsy of Vastus lateralis, preparation of purified mitochondria, and respirometric profiling are described. The use of small muscle volume makes this technique suitable for clinical research applications.

経皮針生検によって得られた骨格筋組織から単離されたミトコンドリアの調製と呼吸計測評価

Respirometric profiling of isolated mitochondria is commonly used to investigate electron transport chain function. We describe a method for obtaining ...

Semi-automated Analysis of Mouse Skeletal Muscle Morphology and Fiber-type Composition

For years, distinctions between skeletal muscle fiber types were best visualized by myosin-ATPase staining. More recently, immunohistochemical staining of myosin heavy chain (MyHC) isoforms has emerged as a finer discriminator of fiber-type. Type I, type IIA, type IIX and type IIB fibers can now be identified with precision based on their MyHC profile; however, manual analysis of these data can be slow and down-right tedious. In this regard, rapid, accurate assessment of fiber-type composition and morphology is a very desirable tool. Here, we present a protocol for state-of-the-art immunohistochemical staining of MyHCs in frozen sections obtained from mouse hindlimb muscle in concert with a novel semi-automated algorithm that accelerates analysis of fiber-type and fiber morphology. As expected, the soleus muscle displayed staining for type I and type IIA fibers, but not for type IIX or type IIB fibers. On the other hand, the tibialis anterior muscle was composed predominantly of type IIX and type IIB fibers, a small fraction of type IIA fibers and little or no type I fibers. Several image transformations were used to generate probability maps for the purpose of measuring different aspects of fiber morphology (i.e., cross-sectional area (CSA), maximal and minimal Feret diameter). The values obtained for these parameters were then compared with manually-obtained values. No significant differences were observed between either mode of analysis with regards to CSA, maximal or minimal Feret diameter (all p &#62; 0.05), indicating the accuracy of our method. Thus, our immunostaining analysis protocol may be applied to the investigation of effects on muscle composition in many models of aging and myopathy.

Immunohistochemical staining of myosin heavy chain isoforms has emerged as the state-of-the-art discriminator of skeletal muscle fiber-type (i.e., type I, type IIA, type IIX, type IIB). Here, we present a staining protocol along with a novel semi-automated algorithm that facilitates rapid assessment of fiber-type and fiber morphology.

マウス骨格筋形態と繊維の種類組成の半自動解析

For years, distinctions between skeletal muscle fiber types were best visualized by myosin-ATPase staining. More recently, immunohistochemical staining of ...

Nerve Stimulator-guided Injection of Autologous Stem Cells Near the Equine Left Recurrent Laryngeal Nerve

Recurrent laryngeal neuropathy (RLN) commonly affects horses and is characterized by abnormal respiratory sounds and exercise intolerance. The recurrent laryngeal nerve shows lesions of demyelination. The benefit of applying stem cells to demyelinated nerves has been demonstrated in various animal models. The aim of the study was to test the feasibility and safety of a peri-neuronal injection of autologous muscle-derived mesenchymal stem cells to the left recurrent laryngeal nerve in healthy horses by using an electrical nerve stimulator.
Muscle-derived stems cell are obtained from five healthy Standardbred horses by sampling 20 mg of muscle tissue with a semi-automatic 14 G biopsy needle from the triceps muscle. Movements of the larynx are monitored via upper-airway video endoscopy. The left recurrent laryngeal nerve is approached with an insulated nerve block needle. Nerve stimulation is applied, starting at 2 mA, and the successful abduction of the left arytenoid is monitored. The stimulation intensity is reduced progressively. When a loss of the motor response is observed at 0.5 mA, 107 autologous muscle-derived stem cells are injected. Two examiners, who are blinded to the time point, score the laryngeal function of the horses prior to the treatment and at day 1, day 7, and day 28 after the injection of the cells. In a sixth horse, 1 mL of 2% lidocaine is injected to further confirm the correct positioning of the needle. This leads to a temporary paralysis of the left arytenoid cartilage.
This study proves that the recurrent laryngeal nerve can be approached with the help of an electrical nerve stimulator and that the electrical stimulation of the nerve is well tolerated by the horses. No modification of the laryngeal function was observed in any of the horses after the injection of the stem cells. Further studies should be conducted to describe the effects of a peri-neuronal injection of autologous muscle-derived mesenchymal stem cells to horses suffering from RLN.

Here, we present a protocol to inject autologous muscle-derived stem cells in the proximity to the recurrent laryngeal nerve with the help of an electrical nerve stimulator. This novel technique may become useful for the treatment of equine recurrent laryngeal neuropathy.

神経刺激誘導馬の左反回神経に近い自家幹細胞注入

Recurrent laryngeal neuropathy (RLN) commonly affects horses and is characterized by abnormal respiratory sounds and exercise intolerance. The recurrent ...

Immunolabelling Myofiber Degeneration in Muscle Biopsies

The necrosis of muscle fibres (myonecrosis) plays a central role in the pathogenesis of several muscle conditions, including muscular dystrophies. Therapeutic options addressing the causes of muscular dystrophy pathogenesis are expected to alleviate muscle degeneration. Therefore, a method to assay and quantify the extent of cell death in muscle biopsies is needed. Conventional methods to observe myofiber degeneration in situ are either poorly quantitative or rely on the injection of vital dyes. In this article, an immunofluorescence protocol is described that stains necrotic myofibers by targeting immunoglobulin G (IgG) uptake by myofibers. The IgG uptake method is based on cell features characterizing the necrotic demise, including 1) the loss of plasma membrane integrity with the release of damage-associated molecular patterns and 2) the uptake of plasmatic proteins. In murine cross-sections, the co-immunolabelling of myofibers, extracellular matrix proteins, and mouse IgG allows clean and straightforward identification of myofibers with necrotic fate. This simple method is suitable for quantitative analysis and applicable to all species, including human samples, and does not require the injection of vital dye. The staining of necrotic myofibers by IgG uptake can also be paired with other co-immunolabelling.

Described here is a protocol for direct immunolabelling of necrotic myofibers in muscle cryosections. Necrotic cells are permeable to serum proteins, including immunoglobulin G (IgG). Revealing the uptake of IgG by myofibers allows the identification and quantification of myofibers that undergo necrosis regardless of muscle condition.

筋生検における免疫標識筋線維変性

The necrosis of muscle fibres (myonecrosis) plays a central role in the pathogenesis of several muscle conditions, including muscular dystrophies. ...

生物学

Collection of Skeletal Muscle Biopsies from the Superior Compartment of Human Musculus Tibialis Anterior for Mechanical Evaluation

The mechanical properties of contracting skeletal fibers are crucial indicators of overall muscle health, function, and performance. Human skeletal muscle biopsies are often collected for these endeavors. However, relatively few technical descriptions of biopsy procedures, outside of the commonly used musculus vastus lateralis, are available. Although the biopsy techniques are often adjusted to accommodate the characteristics of each muscle under study, few technical reports share these changes to the greater community. Thus, muscle tissue from human participants is often wasted as the operator reinvents the wheel. Expanding the available material on biopsies from a variety of muscles can reduce the incident of failed biopsies. This technical report describes a variation of the modified Bergstr&#246;m technique on the musculus tibialis anterior that limits fiber damage and provides fiber lengths adequate for mechanical evaluation. The surgery is an outpatient procedure that can be completed in an hour. The recovery period for this procedure is immediate for light activity (i.e., walking), up to three days for the resumption of normal physical activity, and about one week for wound care. The extracted tissue can be used for mechanical force experiments and here we present representative activation data. This protocol is appropriate for most collection purposes, potentially adaptable to other skeletal muscles, and may be improved by modifications to the collection needle.

This technical report describes a variation of the modified Bergstr&#246;m technique for the biopsy of the musculus tibialis anterior that limits fiber damage.

機械評価のためのヒト筋学の上室から骨格筋生検の収集

The mechanical properties of contracting skeletal fibers are crucial indicators of overall muscle health, function, and performance. Human skeletal muscle ...

Isolation of Mitochondria from Mouse Skeletal Muscle for Respirometric Assays

Most of the cell&#39;s energy is obtained through the degradation of glucose, fatty acids, and amino acids by different pathways that converge on the mitochondrial oxidative phosphorylation (OXPHOS) system, which is regulated in response to cellular demands. The lipid molecule Coenzyme Q (CoQ) is essential in this process by transferring electrons to complex III in the electron transport chain (ETC) through constant oxidation/reduction cycles. Mitochondria status and, ultimately, cellular health can be assessed by measuring ETC oxygen consumption using respirometric assays. These studies are typically performed in established or primary cell lines that have been cultured for several days. In both cases, the respiration parameters obtained may have deviated from normal physiological conditions in&#160;any given organ or tissue.
Additionally, the intrinsic characteristics of cultured single fibers isolated from skeletal muscle impede this type of analysis. This paper presents an updated and detailed protocol for the analysis of respiration in freshly isolated mitochondria from mouse skeletal muscle. We also provide solutions to potential problems that could arise at any step of the process. The method presented here could be applied to compare oxygen consumption rates in diverse transgenic mouse models and study the mitochondrial response to drug treatments or other factors such as aging or sex. This is a feasible method to respond to crucial questions about mitochondrial bioenergetics metabolism and regulation.

Here, we describe a detailed method for mitochondria isolation from mouse skeletal muscle and the subsequent analysis of respiration by Oxygen Consumption Rate (OCR) using microplate-based respirometric assays. This pipeline can be applied to study the effects of multiple environmental or genetic interventions on mitochondrial metabolism.

呼吸法アッセイのためのマウス骨格筋からのミトコンドリアの単離

Most of the cell&#39;s energy is obtained through the degradation of glucose, fatty acids, and amino acids by different pathways that converge on the ...

人間

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