Name: collection of skeletal muscle biopsies from the superior compartment of human musculus tibialis anterior for mechanical evaluation
Uploaded: 2020-09-27T14:30:00
Description: Watch this Scientific Journal Video about Collection of Skeletal Muscle Biopsies from the Superior Compartment of Human Musculus Tibialis Anterior for Mechanical Evaluation at JoVE.com

我们感谢米夏拉·劳、莉娅-费迪亚·里斯曼、迈克尔·马什、贾尼娜-索菲·滕勒、基里安·金梅斯坎普和沃尔夫冈·林克协助该项目。该项目的资金由MERSUR基金会（ID：An-2016-0050）提供给卫生署。

注意：下面，我们概述了一个协议，从参加单独正在进行的研究的志愿者的 TA 中收获机械未损坏的纤维。该协议与Shanely等人描述的相似，他们描述了经过的《大法》中经过的改进的伯格斯特伦技术。此处提供的信息已被我们的研究小组改进，但可能并非适合所有实验室组或组织设置。我们只给出指南，强烈建议新进入活检收集的实验室在尝试任何人体试验之前咨询有经验的实?...

<ol>
	<li>Franchi, M., 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=Architectural,+functional+and+molecular+responses+to+concentric+and+eccentric+loading+in+human+skeletal+muscle.">Architectural, functional and molecular responses to concentric and eccentric loading in human skeletal muscle.</a> Acta Physiologica. 210 (3), 642-654 (2014).</li><li>Duchene, G. B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=De+la+paralysie+musculaire+pseudo-hypertrophique,+ou+paralysie+myo-scl&#233;rosique+/+par+le+Dr+Duchenne+(de+Boulogne).">De la paralysie musculaire pseudo-hypertrophique, ou paralysie myo-scl&#233;rosique / par le Dr Duchenne (de Boulogne).</a> Archives of General Internal Medicine. 11 (30), (1868).</li><li>Shanely, R. A., 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=Human+skeletal+muscle+biopsy+procedures+using+the+modified+Bergstr&#246;m+technique.">Human skeletal muscle biopsy procedures using the modified Bergstr&#246;m technique.</a> Journal of Visualized Experiments. (91), e51812 (2014).</li><li>Evans, W. J., Phinney, S. D., Young, V. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Suction+applied+to+a+muscle+biopsy+maximizes+sample+size.">Suction applied to a muscle biopsy maximizes sample size.</a> Medicine and Science in Sports and Exercise. 14 (1), 101-102 (1982).</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=Percutaneous+needle+biopsy+of+skeletal+muscle+in+physiological+and+clinical+research.">Percutaneous needle biopsy of skeletal muscle in physiological and clinical research.</a> Scandinavian Journal of Clinical and Laboratory Investigation. 35 (7), 609-616 (1975).</li><li>Baczynska, A. M., 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=Human+Vastus+Lateralis+Skeletal+Muscle+Biopsy+Using+the+Weil-Blakesley+Conchotome.">Human Vastus Lateralis Skeletal Muscle Biopsy Using the Weil-Blakesley Conchotome.</a> Journal of Visualized Experiments. (109), e53075 (2016).</li><li>Pesta, D., Gnaiger, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-resolution+respirometry:+OXPHOS+protocols+for+human+cells+and+permeabilized+fibers+from+small+biopsies+of+human+muscle.">High-resolution respirometry: OXPHOS protocols for human cells and permeabilized fibers from small biopsies of human muscle.</a> Methods in Molecular Biology. 810, 25-58 (2012).</li><li>Buck, E., 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=High-resolution+respirometry+of+fine-needle+muscle+biopsies+in+pre-manifest+Huntington's+disease+expansion+mutation+carriers+shows+normal+mitochondrial+respiratory+function.">High-resolution respirometry of fine-needle muscle biopsies in pre-manifest Huntington's disease expansion mutation carriers shows normal mitochondrial respiratory function.</a> Plos One. 12 (4), 01175248 (2017).</li><li>Murgia, M., 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=Single+Muscle+Fiber+Proteomics+Reveals+Fiber-Type-Specific+Features+of+Human+Muscle+Aging.">Single Muscle Fiber Proteomics Reveals Fiber-Type-Specific Features of Human Muscle Aging.</a> Cell Reports. 19 (11), 2396-2409 (2017).</li><li>Friedmann-Bette, B., 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=Effects+of+strength+training+with+eccentric+overload+on+muscle+adaptation+in+male+athletes.">Effects of strength training with eccentric overload on muscle adaptation in male athletes.</a> European Journal of Applied Physiology. 108 (4), 821-836 (2010).</li><li>McPhee, J. S., 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=The+contributions+of+fibre+atrophy,+fibre+loss,+in+situ+specific+force+and+voluntary+activation+to+weakness+in+sarcopenia.">The contributions of fibre atrophy, fibre loss, in situ specific force and voluntary activation to weakness in sarcopenia.</a> The Journals of Gerontology. Series A, Biological Sciences and Medical Sciences. 73 (10), 1287-1294 (2018).</li><li>Nocella, M., Cecchi, G., Bagni, M. A., Colombini, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Force+enhancement+after+stretch+in+mammalian+muscle+fiber:+no+evidence+of+cross-bridge+involvement.">Force enhancement after stretch in mammalian muscle fiber: no evidence of cross-bridge involvement.</a> American Journal of Physiology. Cell Physiology. 307 (12), 1123-1129 (2014).</li><li>Patel, J. R., McDonald, K. S., Wolff, M. R., Moss, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ca2++binding+to+troponin+C+in+skinned+skeletal+muscle+fibers+assessed+with+caged+Ca2++and+a+Ca2++fluorophore.+Invariance+of+Ca2++binding+as+a+function+of+sarcomere+length.">Ca2+ binding to troponin C in skinned skeletal muscle fibers assessed with caged Ca2+ and a Ca2+ fluorophore. Invariance of Ca2+ binding as a function of sarcomere length.</a> The Journal of Biological Chemistry. 272 (9), 6018-6027 (1997).</li><li>Hessel, A. L., Joumaa, V., Eck, S., Herzog, W., Nishikawa, K. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimal+length,+calcium+sensitivity+and+twitch+characteristics+of+skeletal+muscles+from+mdm+mice+with+a+deletion+in+N2A+titin.">Optimal length, calcium sensitivity and twitch characteristics of skeletal muscles from mdm mice with a deletion in N2A titin.</a> The Journal of Experimental Biology. 222, (2019).</li><li>Joumaa, V., Herzog, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calcium+sensitivity+of+residual+force+enhancement+in+rabbit+skinned+fibers.">Calcium sensitivity of residual force enhancement in rabbit skinned fibers.</a> American Journal of Physiology. Cell Physiology. 307 (4), 395-401 (2014).</li><li>Joumaa, V., Rassier, D. E., Leonard, T. R., Herzog, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+origin+of+passive+force+enhancement+in+skeletal+muscle.">The origin of passive force enhancement in skeletal muscle.</a> American Journal of Physiology. Cell Physiology. 294 (1), 74-78 (2008).</li><li>Hilber, K., Galler, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanical+properties+and+myosin+heavy+chain+isoform+composition+of+skinned+skeletal+muscle+fibres+from+a+human+biopsy+sample.">Mechanical properties and myosin heavy chain isoform composition of skinned skeletal muscle fibres from a human biopsy sample.</a> Pflugers Archiv: European Journal of Physiology. 434 (5), 551-558 (1997).</li><li>Miller, M. S., 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=Chronic+heart+failure+decreases+cross-bridge+kinetics+in+single+skeletal+muscle+fibres+from+humans.">Chronic heart failure decreases cross-bridge kinetics in single skeletal muscle fibres from humans.</a> The Journal of Physiology. 588, 4039-4053 (2010).</li><li>Pinnell, R. A. M., 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=Residual+force+enhancement+and+force+depression+in+human+single+muscle+fibres.">Residual force enhancement and force depression in human single muscle fibres.</a> Journal of Biomechanics. 91, 164-169 (2019).</li><li>Einarsson, F., Runesson, E., Frid&#233;n, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Passive+mechanical+features+of+single+fibers+from+human+muscle+biopsies--effects+of+storage.">Passive mechanical features of single fibers from human muscle biopsies--effects of storage.</a> Journal of Orthopaedic Surgery and Research. 3, 22 (2008).</li><li>Flann, K. L., LaStayo, P. C., McClain, D. A., Hazel, M., Lindstedt, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscle+damage+and+muscle+remodeling:+no+pain,+no+gain.">Muscle damage and muscle remodeling: no pain, no gain.</a> The Journal of Experimental Biology. 214, 674-679 (2011).</li><li>Commission for Hospital Hygiene and Infection Prevention (KRINKO), Federal Institute for Drugs and Medical Devices (BfArM). <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anforderungen+an+die+Hygiene+bei+der+Aufbereitung+von+Medizinprodukten+[Hygiene+requirements+for+the+reprocessing+of+medical+devices].">Anforderungen an die Hygiene bei der Aufbereitung von Medizinprodukten [Hygiene requirements for the reprocessing of medical devices].</a> Bundesgesundheitsblatt, Gesundheitsforschung, Gesundheitsschutz. 55 (10), 1244-1310 (2012).</li><li>Koch-Institut, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Erg&#228;nzung+zur+Empfehlung+Anforderungen+an+die+Hygiene+bei+der+Aufbereitung+von+Medizinprodukten.">Erg&#228;nzung zur Empfehlung Anforderungen an die Hygiene bei der Aufbereitung von Medizinprodukten.</a> RKI-Bib1. , (2018).</li><li>Rutala, W. A., Weber, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disinfection+and+sterilization+in+healthcare+facilities.">Disinfection and sterilization in healthcare facilities.</a> Practical Healthcare Epidemiology. , 58-81 (2018).</li><li>Rassier, D. E., MacIntosh, B. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sarcomere+length-dependence+of+activity-dependent+twitch+potentiation+in+mouse+skeletal+muscle.">Sarcomere length-dependence of activity-dependent twitch potentiation in mouse skeletal muscle.</a> BMC Physiology. 2, 19 (2002).</li><li>Mounier, Y., Holy, X., Stevens, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Compared+properties+of+the+contractile+system+of+skinned+slow+and+fast+rat+muscle+fibres.">Compared properties of the contractile system of skinned slow and fast rat muscle fibres.</a> Pflugers Archiv: European Journal of Physiology. 415 (2), 136-141 (1989).</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 Neurologica Scandinavica. 59 (6), 317-323 (1979).</li><li>Dietrichson, P., 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=Conchotome+and+needle+percutaneous+biopsy+of+skeletal+muscle.">Conchotome and needle percutaneous biopsy of skeletal muscle.</a> Journal of Neurology, Neurosurgery, and Psychiatry. 50 (11), 1461-1467 (1987).</li><li>Iachettini, S., 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=Tibialis+anterior+muscle+needle+biopsy+and+sensitive+biomolecular+methods:+a+useful+tool+in+myotonic+dystrophy+type+1.">Tibialis anterior muscle needle biopsy and sensitive biomolecular methods: a useful tool in myotonic dystrophy type 1.</a> European Journal of Histochemistry. 59 (4), 2562 (2015).</li><li>Cotter, J. A., 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=Suction-modified+needle+biopsy+technique+for+the+human+soleus+muscle.">Suction-modified needle biopsy technique for the human soleus muscle.</a> Aviation, Space, and Environmental Medicine. 84 (10), 1066-1073 (2013).</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 &amp; Nerve. 6 (9), 676-683 (1983).</li><li>Gibreel, W. O., 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=Safety+and+yield+of+muscle+biopsy+in+pediatric+patients+in+the+modern+era.">Safety and yield of muscle biopsy in pediatric patients in the modern era.</a> Journal of Pediatric Surgery. 49 (9), 1429-1432 (2014).</li><li>Cuisset, J. M., 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=Muscle+biopsy+in+children:+Usefulness+in+2012.">Muscle biopsy in children: Usefulness in 2012.</a> Revue Neurologique. 169 (8-9), 632-639 (2013).</li><li>Nilipor, Y., 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=Evaluation+of+one+hundred+pediatric+muscle+biopsies+during+a+2-year+period+in+mofid+children+and+toos+hospitals.">Evaluation of one hundred pediatric muscle biopsies during a 2-year period in mofid children and toos hospitals.</a> Iranian Journal of Child Neurology. 7 (2), 17-21 (2013).</li><li>Schiaffino, S., Reggiani, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fiber+types+in+mammalian+skeletal+muscles.">Fiber types in mammalian skeletal muscles.</a> Physiological Reviews. 91 (4), 1447-1531 (2011).</li><li>Wang, K., Wright, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Architecture+of+the+sarcomere+matrix+of+skeletal+muscle:+immunoelectron+microscopic+evidence+that+suggests+a+set+of+parallel+inextensible+nebulin+filaments+anchored+at+the+Z+line.">Architecture of the sarcomere matrix of skeletal muscle: immunoelectron microscopic evidence that suggests a set of parallel inextensible nebulin filaments anchored at the Z line.</a> The Journal of Cell Biology. 107 (6), 2199-2212 (1988).</li><li>Ma, W., Gong, H., Irving, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myosin+head+configurations+in+resting+and+contracting+murine+skeletal+muscle.">Myosin head configurations in resting and contracting murine skeletal muscle.</a> International Journal of Molecular Sciences. 19 (9), (2018).</li><li>Ma, W., Gong, H., Kiss, B., Lee, E. J., Granzier, H., Irving, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thick-Filament+Extensibility+in+Intact+Skeletal+Muscle.">Thick-Filament Extensibility in Intact Skeletal Muscle.</a> Biophysical Journal. 115 (8), 1580-1588 (2018).</li><li>Bonafiglia, J. T., 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=A+comparison+of+pain+responses,+hemodynamic+reactivity+and+fibre+type+composition+between+Bergstr&#246;m+and+microbiopsy+skeletal+muscle+biopsies.">A comparison of pain responses, hemodynamic reactivity and fibre type composition between Bergstr&#246;m and microbiopsy skeletal muscle biopsies.</a> Current Research in Physiology. 3, 1-10 (2020).</li><li>Wickiewicz, T. L., Roy, R. R., Powell, P. L., Edgerton, V. 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+architecture+of+the+human+lower+limb.">Muscle architecture of the human lower limb.</a> Clinical Orthopaedics and Related Research. (179), 275-283 (1983).</li></ol>

在这份报告中，我们描述了一种从TA中对结构上未受损的肌肉组织进行活检的技术。我们发现，这个程序产生一个可接受的含量的可用肌肉纤维（5-10纤维束制剂每50毫克收集的组织）机械测试。此外，我们有足够的组织进行后续的机械、遗传和蛋白质遗传学实验。
有几个方法通常用于收集肌肉活,检3，4，6，27，28。4,6</su...

为临床或研究目的研究人类肌肉生理学往往需要肌肉活检。例如，人类肌肉生理学和生物力学的一个主要挑战是区分和理解肌肉性能对运动的各种适应。性能适应不仅包括结构适应（例如，收缩蛋白的变化，肌肉结构），还包括神经适应1，这是很难，如果不是不可能，单独评估时，测试完好无损的原位人类肌肉。纤维级实验去除这些高阶成分，并允许更直接地评估肌肉收缩，并可通过活检技术收集。肌肉活检已经收集至少自1868年2。今天，收集肌肉活检的主要技术是修改的Bergström技术3,3，4，5，,5虽然其他技术可用，包括使用威尔-布莱克斯利孔胸6或所谓的细针7,7，8。所有这些技术都使用特殊的针状仪器，这些仪器被设计成进入肌肉并切割一块组织。具体来说，经过修改的Bergström技术使用大改性针头（此处为 5 mm 针头尺寸）;图1）有一个靠近针尖的窗户和一个较小的内部小刀，在针尖上和向下移动，在穿过针尖窗口时切割肌肉。在这个神圣的小车内是一个拉罗德，向上和向下移动的轴的特罗卡，并推动活检到针窗。要将肌肉拉入针窗，请连接一根吸气软管，吸出针头的空气，通过负压将肌肉拉入针窗。
肌肉活检通常是为了研究蛋白质含量、基因表达或疾病引起的形态变化，或对,运动计划,1、9、10、11,9的反应而获得的。10肌肉活检的另一个关键用途是机械实验，如测量纤维收缩力，肌肉纤维僵硬，和历史依赖肌肉属性12，13，14，15，16。12,13,14,15,16单光纤或光纤束力学通过将光纤连接在长度电机和力传感器之间，在控制光纤长度同时测量力的专用钻机上进行测量。通过渗透（如皮肤）纤维，肉瘤膜变得渗透到沐浴液中的化学物质，允许通过改变钙浓度进行活化控制。此外，通过将有关试剂添加到沐浴液中，可以很容易地评估收缩特性对化学品/药物/其他蛋白质的影响。然而，虽然这项技术在其他动物模型中被高度使用，但对人类肌肉活检17、18、19,的皮片纤维进行机械测试的研究明显较少。原因之一是活检工具和协议旨在去除尽可能多的肌肉组织，而较少考虑组织提取过程中遭受的结构损伤程度。事实上，最近的活检方案建议推动活检针进入肌肉，并收集2-4块肌肉3。这个过程本身对DNA或蛋白质物质的损害很小，但往往破坏纤维和沙康结构，使肌肉纤维的激活变得不稳定或不可能。此外，活检中纤维的相对长度通常很短（<2 mm），不容易用于机械测试。对于机械测试，理想的纤维长（3-5 毫米），且没有结构损坏。
更先进的组织提取技术可用于限制纤维损伤。例如，一组20 人利用了先前规划的前臂"开放手术"（例如骨折修复），其中肌肉完全暴露，外科医生能够可视化肌肉结构，并仔细解剖肌肉组织相对较大和结构上未损坏的样本（15毫米x5毫米x5毫米）。这种"开放活检"技术在参与者正在接受先前计划的程序时受到青睐，因此限制了潜在参与者的库，尤其是对健康成年人，否则不会进行手术。因此，许多为研究目的进行的活检都是作为门诊程序进行的，切口部位尽可能小，以限制感染风险、疤痕和愈合时间。因此，大多数活检 是盲目 收集的（即，操作者无法看到收集针头，因为它通过筋膜进入肌肉）。这意味着活检的质量几乎完全基于操作者的技能和经验。每个肌肉在收集组织时都有自己的困难，例如有违反神经和血管的风险，选择理想的收集深度和位置，并决定一个合适的身体位置，以保持肌肉尽可能松弛。不幸的是，大多数肌肉特异性技能集没有写下来，所以每个医生必须"重新发明车轮"时，执行活检的肌肉新到他们。这种缺乏经验通常导致几个收集低质量，直到医生确定对肌肉的活检的最佳做法。新手医生经常通过与经验更丰富的同事交谈来学习这种技能，但相对而言，有关这一问题的信息和同行评议文本相对较少，尤其是对于传统上不用于活检收集的肌肉。如果我们考虑上述信息，以及招募人类志愿者进行活检的困难，显然需要更多的教学信息，从而最大限度地提高每个参与者的成功机会。
因此，本文的目的是提出一种肌肉活检技术，为成功采集肌肉活检和长而未损坏的纤维片段进行机械测试提供了方案。人类肌肉活检通常是进行，和活检训练材料的大部分是，肌肉广大后。其相对较大的肌肉大小和表面位置相对于皮肤允许收集足够的肌肉组织，同时尽量减少病人的不适和身体创伤,1，21。然而，在纵向训练研究中使用广大后法存在一些局限性。例如，在包括培训计划的实验协议期间，学员必须在通常为期 2-6 个月的学习期间内避免在学习之外进行额外培训。对于运动员来说，这通常是不可能的，因为广大的后天运动员通常在典型的练习（如蹲下、跳跃）期间进行训练，或者通常用于这项运动（例如跑步、骑自行车）。这些远离研究目的的单独训练经验可能导致肌肉适应，改变肌肉机制、结构以及生理学，使难以或不可能知道研究的实验协议对肌肉特性的真正影响。对于这些类型的研究，最好选择目标肌肉，这往往不是训练团的重点。肌肉头骨前 （TA） 是满足上述要求的理想目标肌肉。此外，培训干预措施可以针对 TA 使用可控的方法，例如使用测功机。几乎没有与 TA 肌肉活检相关的训练材料。因此，我们开发了一个修改的协议，从TA收集相对未受损的肌肉活检。

<table>
	<tbody>
		<tr>
			<td>26 guage subcutaneous needle with 2 ml glass syringe</td>
			<td>B. Braun Melsungen AG 
			Carl-Braun-Stra&#223;e 1 
			34212 Melsungen, Hessen 
			Germany 
			&#160;</td>
			<td>4606027V</td>
			<td>Drug administration</td>
		</tr>
		<tr>
			<td>5mm Berst&#246;m needle</td>
			<td>homemade</td>
			<td>N/A</td>
			<td>Tissue collection. Similar to other Berst&#246;m needles</td>
		</tr>
		<tr>
			<td>Acrylastic</td>
			<td>BSN medical GmbH 
			22771 Hamburg</td>
			<td>269700</td>
			<td>elastic compression bandage</td>
		</tr>
		<tr>
			<td>Complete protease inhibitor cocktail</td>
			<td>Roche Diagnostics, Mannheim, Germany</td>
			<td>11836145001</td>
			<td>Protease inhibitor tabeletes added to all solutions that hold muscle tissue.</td>
		</tr>
		<tr>
			<td>Cutasept</td>
			<td>PAUL HARTMANN AG 
			Paul-Hartmann-Stra&#223;e 12 
			89522 Heidenheim 
			Germany</td>
			<td>9805630</td>
			<td>Disenfectant spray for the skin</td>
		</tr>
		<tr>
			<td>Leucomed T plus</td>
			<td>BSN medical GmbH 
			22771 Hamburg</td>
			<td>7238201</td>
			<td>Transparent wound dressing with wound pad to seal the wound and protect against infection</td>
		</tr>
		<tr>
			<td>Leukostrip</td>
			<td>Smith and Nephew medical Limitied 101 Hessle road, 
			Hull 
			Great Britain</td>
			<td>66002876</td>
			<td>wound closure</td>
		</tr>
		<tr>
			<td>Surgical disposable scalpels</td>
			<td>Aesculap AG 
			Am Aesculap-Platz 
			78532 Tuttlingen 
			Germany</td>
			<td>BA200 series</td>
			<td>Incision</td>
		</tr>
		<tr>
			<td>Unihaft cohesive elastic bandage</td>
			<td>BSN medical GmbH 
			22771 Hamburg</td>
			<td>4589600</td>
			<td>cohesive elastic bandage that protects against mechanical impact</td>
		</tr>
		<tr>
			<td>Xylocitin 2% with Epinephrin</td>
			<td>Milbe GmbH 
			M&#252;nchner Stra&#223;e 15 
			06796 Brehna 
			Germany</td>
			<td>N/A</td>
			<td>Controlled substance anesthesia, vasoconstriction</td>
		</tr>
	</tbody>
</table>

collection of skeletal muscle biopsies from the superior compartment of human musculus tibialis anterior for mechanical evaluation

收缩骨骼纤维的机械性能是整体肌肉健康、功能和性能的关键指标。人类骨骼肌活检经常收集为这些努力。然而，除了常用的粘液后，活检程序的技术描述相对较少。虽然活检技术经常进行调整，以适应每个肌肉在研究的特点，很少有技术报告分享这些变化，以更大的社区。因此，当操作员重新发明车轮时，人类参与者的肌肉组织经常被浪费掉。从各种肌肉中扩展活检的可用材料可以减少活检失败的事件。本技术报告描述了改进后的 Bergström 技术在粘液头骨前的变化，该技术可限制纤维损坏并提供足够的纤维长度进行机械评估。手术是一个门诊手术，可以在一小时内完成。此程序的恢复期是立即为轻活动（即步行），最多三天恢复正常身体活动，约一个星期的伤口护理。提取的组织可用于机械力实验，我们在这里提供具有代表性的激活数据。该协议适用于大多数收集目的，可能适应其他骨骼肌，并可能通过修改收集针进行改进。

参与者的整个时间承诺是大约1小时（10分钟会诊，10分钟超声波，20分钟的手术准备和麻醉剂，10分钟的手术和10分钟的恢复）。通常，参与者不自觉地激活他们的TA，需要一致的提醒，以保持肌肉尽可能放松。当活检针在肌肉内时，参与者通常会报告活检针周围区域有独特的"压力"感觉，偶尔会出现中度到剧烈的不适。一次，参与者的脚趾在手术过程中稍微抽筋，但在针头被取出后立即停止。活检?...

Watch this Scientific Journal Video about Collection of Skeletal Muscle Biopsies from the Superior Compartment of Human Musculus Tibialis Anterior for Mechanical Evaluation at JoVE.com

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

本技术报告描述了经过改良的Bergström技术的变化，用于对前骨质纤维的活检，以限制纤维损伤。

从人类肌肉提比利斯前部高级隔间收集骨骼肌肉活检，用于机械评估

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

欢迎来到鲁尔大学波鸿体育科学学院。今天，我们将演示从人类肌肉前头骨的上级隔间收集的骨骼肌活检。虽然活检技术在收集肌肉组织方面是有效的，但纤维在结构上经常受到损伤，因此很难评估纤维力学。我们提出了一种改进的Bergstrom技术的变化，该技术可以限制纤维损伤并最大限度地延长纤维长度，从而可以对人纤维进行机械评估。演示将由队医马库斯·德·马里斯博士和研究科学家丹尼尔·哈恩博士和安东尼·赫塞尔进行。参与者和医生首先开会审查和讨论程序。如果满意，参与者签署自愿知情同意合同。指示主体躺下，保持肢体尽可能放松。Dorsiflex 参与者的脚， 使头骨前是松弛的。使用超声波探针可视化前肌肉腹部内表面隔间的反面，识别没有主要动脉或神经的活检区域。最后，记录中央动脉瘤的深度，使收集针永远不会被驱动超过这一点。旋转超声波探针以访问已识别目标区域的下垂平面。使用可见分册保存肌肉体积的图像。手术工具在手术前进行消毒和准备，并在书面协议中全面描述。伯格斯特伦针有一个窗口接近尖端。在针头内是一个较小的空心小车，它上下移动轴和削减肌肉时，通过针窗。在小车内是一个较小的公羊，用于将活检从小车中推出。要将肌肉拉入针窗，吸气软管附着在针头顶部，吸气管从针头中吸出空气，并通过负压将肌肉从针窗中拉出。指示学员在手术台上放上小手术台，放松腿部肌肉。复制超声波成像期间使用的轻微脚踝多西反射，然后固定脚踝关节，以尽量减少手术期间的旋转。剃须和消毒目标切口区域。接下来，用肾上腺素进行1.5个PC的2%xylocitin的肤浅注射。这作为局部麻醉剂和血管收缩器。影响时间是 20 到 30 分钟。用屏幕将患者的视线移除到活检位置。在先前标记的活检部位，用一个无菌的手术刀切开皮肤和筋膜，露出底层肌肉组织，使一厘米近到离骨切口。将活检针放置在切口上，并垂直于皮肤。将活检针推入肌肉半到一厘米。针头运动可能有一些光阻力，但任何额外的阻力是不正常的，通常表示切口不正常。接下来将针头定向到近近位置，将针头再推入一到两厘米的肌肉。针头的确切方向和行程深度将是参与者特定的。从而基于超声波检查。将注射器软管连接到活检针上，并在针头中产生负压。这将通过针窗绘制肌肉组织。强行将小车推到针窗上切除肌肉。从腿上取出针头，并传递给技术人员进行组织处理。使用伤口封闭胶带或粘合剂主要关闭伤口，并涂抹敷料，以保护伤口免受感染或机械磨损。指示学员站起来四处走动。他们会很痛，但能马上四处走动。要从活检窗口取出组织，首先将组织暴露，然后将针头浸入充满生理缓冲液的邪恶中，并大力移动针头，直到肌肉从针头中浮出，进入溶液中。使用标准方法处理肌肉组织进行机械评估。我们最近从使用此协议的参与者那里收集了 22 次活检。平均收集产量为100至200毫克可用肌肉组织。每次活检产生150至300个长度为3至5毫米的未损坏纤维。纤维束用甘油进行化学剥皮，然后在平均长度为2.7微米的高钙溶液中激活。正确储存的活检在几个月内可以进行机械测试。这个活检程序需要一个小时。活检并发症非常罕见，但很可能是切口部位的感染。人类肌肉生理学和生物力学的一个主要挑战是区分肌肉性能对运动和疾病的各种适应。为了适当确定纤维水平适应运动或疾病，活检是适当的技术。然而，在组织提取过程中，必须注意防止肌肉损伤。此处演示的协议为这些目的提供了高产量的肌肉组织。

Research

JoVE Journal

Biology

Mitochondrial Isolation from Skeletal Muscle

骨骼肌的线粒体分离

Mitochondria are organelles controlling the life and death of the cell. They participate in key metabolic reactions, synthesize most of the ATP, and ...

科研

生物学

Purification of Progenitors from Skeletal Muscle

纯化的骨骼肌的先觉者

Skeletal muscle contains multiple progenitor populations of distinct embryonic origins and developmental potential. Myogenic progenitors, usually residing ...

Laser-inflicted Injury of Zebrafish Embryonic Skeletal Muscle

激光伤害的斑马鱼胚胎骨骼肌

Various experimental approaches have been used in mouse to induce muscle injury with the aim to study muscle regeneration, including myotoxin injections ...

Isolation and Culture of Individual Myofibers and their Satellite Cells from Adult Skeletal Muscle

单个肌纤维和成人骨骼肌卫星细胞的分离和培养

Muscle  regeneration in the adult is performed by resident stem cells called satellite  cells. Satellite cells are defined by  their position between the ...

Evaluation of Muscle Function of the Extensor Digitorum Longus Muscle Ex vivo and Tibialis Anterior Muscle In situ in Mice

Evaluation of Muscle Function of the Extensor Digitorum Longus Muscle Ex vivo and Tibialis Anterior Muscle In situ in Mice

伸趾长肌的肌肉功能评价<em&gt;体外</em和胫骨前肌<em在原位</em在小鼠

Body  movements are mainly provided by mechanical function of skeletal muscle. Skeletal  muscle is composed of numerous bundles of myofibers that are ...

Assessment of Calcium Sparks in Intact Skeletal Muscle Fibers

钙火花的评估在正常肌肉纤维

Maintaining homeostatic Ca2+ signaling is a fundamental physiological process in living cells. Ca2+ sparks are the elementary units of Ca2+ signaling in ...

Isolation of Blood-vessel-derived Multipotent Precursors from Human Skeletal Muscle

从人骨骼肌隔离血管源性多能前体

Since the discovery of mesenchymal stem/stromal cells (MSCs), the native identity and localization&#160;of MSCs have been obscured by their retrospective ...

Tissue Triage and Freezing for Models of Skeletal Muscle Disease

组织分流和冻结的骨骼肌疾病模型

Skeletal muscle is a unique tissue because of its structure and function, which requires specific protocols for tissue collection to obtain optimal ...