Name: human skeletal muscle biopsy procedures using the modified bergström technique
Uploaded: 2014-09-10T14:00:00.000+00:00
Duration: 7 min 20 s
Description: Watch this Scientific Journal Video about Human Skeletal Muscle Biopsy Procedures Using the Modified BergstrÃƒÂ¶m Technique at JoVE.com

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所描述的骨骼肌过程遵循阿巴拉契亚州立大学伦理审查委员会的指导方针。 注:该小组由操作者（博士或MD训练的活检技术），和至少一个，但理想的是，三个或更多个技术员。操作员负责进行及监督骨骼肌活检过程的各个方面。一技术员（技术员＃1）是立即用于辅助操作者肌肉提取的所有方面负责。这包括在"非无菌手"和施加抽吸，用无菌注射器。第二技师（技师＃2）在主题和按摩主体的对面站的过程中，尽量减少受试者的焦虑谈话从事。第三技师（技师＃3）是负责处理和加工的骨骼肌样品。下游的分析方法决定多少乳房清洁度有理技术人员所需要的加工/制备骨骼肌样品。 （一）除准备<ol><li>收集，准备和组织的肌肉活检程序（ 表1）中指定的材料。去污，消毒BERGSTROM针（外套管和内套管），并根据标准实践16的柱塞。 </li><li>回顾与主体的知情同意书。在同意参加在阿巴拉契亚州立大学使用形式的骨骼肌活检部分在网上补一。 <ol><li>聊聊肌肉活检过程中，重点放在程序的可能存在的风险，备皮，应用麻醉剂，通常感觉的一般方面的问题。指示须仔细阅读同意参加表格并签字确认，这项研究的设计和程序的理解。 </li></ol></li><li>确认标ubject是不过敏的"___坚"型药物（ 如利多卡因）。 </li><li>指示受仰卧在软垫表暴露大腿。将两个一次性吸收护垫与塑料底板主体的腿下。用脚后跟下一个小毛巾卷，使得膝盖完全伸展（升高约1cm），放置在一个轻松的方式的腿部，从而把股外侧肌在缩短的位置。 </li><li>有操作指示受试者对瞬间收缩的露出大腿肌肉，使得活检部位可以显现。注意:股外侧肌活检部位只是前阔筋膜（illiotibial带），大约三分之一的髌骨的顶部和大转子7之间的距离。 </li><li>刚才下面的操作大关（约0.5厘米）的切口部位用细点永久性标记。 </li><li>在操作员确定大致皮褶厚度由PINC兴的皮褶。 </li><li>有运营商和技术人员洗手用肥皂和温水和唐一次性手套。 </li><li>有运营商从区域去除毛〜15公分×15厘米左右的活检部位（经快船）。注意:从活检部位剪切毛发防止头发从进入切口的过程中和结束时，和脱毛允许磁带关闭和皮肤（见3.4）之间的更好的接触。 </li><li>有运营商棉签消毒面积预浸泡局部消毒剂（如碘伏或洗必泰科目碘过敏/贝类）。开始在中心和同心圆朝向剪切区域的外边缘的工作。至少，有一个新的预先浸泡棉签每次重复两次以上。 </li></ol> 2，活检程序<ol><li>有运营商去除一次性手套，用肥皂和温水洗手，并穿上无菌手术GLoves使用无菌技术。 </li><li>有技术＃1存在于无菌窗孔被单给操作者。有运营商，采用无菌技术，将覆盖在活检部位以保持无菌区。 </li><li>有技术人员＃1目前运营商有5毫升注射器配有21G针头，同时保持无菌操作。清洁利多卡因小瓶用酒精棉签的顶端。 <ol><li>有运营商立即撤除5毫升利多卡因。从注射器中取出21克针，并丢弃针头插入锐器容器中。 </li><li>有技术人员＃1目前有一个1英寸半地下25针的操作，同时保持无菌操作。有操作的地方的1半英寸的针入注射器并从注射器排空气泡。 </li></ol></li><li>有技术人员＃1喷氯乙烷在切口部位（约0.5厘米以上，对皮肤的不褪色墨水标记），直到皮肤出现"灼"。 <ol><li>哈已经操作员插入的针大致水平的皮肤进入真皮，抽吸针，然后渗透用〜100μl的利多卡因，以产生一个"气泡"2-4毫米的直径。 <ol><li>有操作抽吸针（略撤回注射器的柱塞），以确认针头尚未放置在血管中。如果血液中出现的注射器，抽出针，丢弃在一个锐器容器，并在2.3重新开始。 </li><li>有操作者预先将针插入皮下组织，抽吸针，然后渗透用〜1毫升利多卡因的组织以形成气泡。一旦水泡已经消退，将在利多卡因装注射器的1½寸毫针垂直插入切口部位，停止向浅筋膜。注:如有可能最初注射利多卡因（类似于蜜蜂蜇）的时感到轻微短暂的刺痛感。 </li></ol></li><li>确保OPERAT或者没有渗透到肌肉与lidocoine因为它是myotoxic 17-19。 </li><li>有运营商抽吸针，然后慢慢注入利多卡因的剩余4毫升同时取出从大腿针。将无菌纱布注射部位，并让受放松，而局部麻醉生效。 </li></ol></li><li>经过2-3分钟，具有技师＃1本操作手术刀，同时保持无菌操作。 <ol><li>有操作者轻轻探查活检部位用手术刀的前端，以确认该区域的麻醉。如果有必要，又注入3-4毫升利多卡因进入切口部位，见2.4.1.2。 </li></ol></li><li>有操作员通过皮肤和皮下组织（〜2-3毫米以上的墨水标记）平行于股骨作出的直1厘米的切口。 <ol><li>有运营商将手术刀通过更深层次的筋膜进入肌肉做一个切口。在每个方向上，一旦做到这一点<html> 。注:如有可能会感到刺痛或压力的更深层次的剪裁，如果手术刀刀片切到肌肉。 </li><li>有技术人员2＃进行谈话的主题和按摩对侧下肢。 </li></ol></li><li>有运营商的地方无菌纱布在切口活检部位宽松的数量，并采用直接压到切口，以减少出血。 <ol><li>有技术＃1的3通金属活栓连接到一次性60毫升注射器和30厘米的延长管（ 图1）。 </li><li>有技术人员＃1连接的锥形塑料管连接器的一端插入延伸管的另一端牢牢地插在锥形塑料管连接器的另一端插入200微升枪头的大底与〜15-18毫米前端切掉。 </li></ol></li></ol>.JPG"WIDTH ="500"/&gt; 图1 BERGSTROM针。 （一）伯格斯坦针（5毫米）的是由（A）外插管及（b）内套管针，相关组件包括:（C）柱塞，（D）200微升枪头〜15-18毫米切断，（五）锥形塑料连接器，（F）3通金属塞阀（克）30厘米延伸管，和（h）一次性60毫升注射器（B）的BERGSTROM针（5毫米）和相关的部件组装而成。（ C）的BERGSTROM针（5毫米）的排出的内套管针约1cm开启外部套管的窗口。 <ol start="9"><li>后出血已消退，具有技师＃1打开高压釜灭菌袋以及呈现5毫米活检针和内套管（ 图1）的操作者，同时保持无菌技术。 <ol><li>有运营商组装针（外套管和内套管针），检查路线和滑动的动作。 </li> &lt;/ OL&gt; </li><li>有操作者牢固地保持针头组件（外套管和内套管针），用双手来防止内套管针旋转。有技术人员＃1牢牢插入剪切枪头插入穿刺针的内套管部分的顶部有轻微的向下扭转运动。确保技术人员＃1不接触操作者或活检针，只枪头。 </li><li>有操作者引入活检针插入通过切口的组织。定位在筋膜的切口（后天"感觉"）与所述活检针的尖端。推进针刚刚过去的筋膜，然后角针向下朝向地板，作为将针推入肌肉。注:受试者可能会遇到一些轻微的不适（ 例如 ，"深压力"，或轻度痉挛感觉），为通过筋膜切口位于与所述活检针的尖端并在针被推进到肌肉。 <ol> &lt;李&gt;有技术员＃2指示须留"宽松"，而不是合同的大腿肌肉，而运营商正在推进针头进入肌肉。 </li></ol></li><li>一旦活检针的位置，有运营商的信号技师＃1，打开一次性60毫升注射器。 <ol><li>有技术人员＃1迅速拉一次性60毫升注射器开到40-50毫升标记活检针中创建的吸力。 </li><li>有操作者拉动内套管出大约1cm，同时保持肌肉内外侧套管针的位置，打开外插管的窗口。有运营商迅速关闭内套管剪切（剪辑），并收集肌肉样本。有运营商的信号技师＃1释放吸力。 </li><li>有技术人员＃1打开活塞房间释放吸力。 </li><li>有操作者旋转所述活检针90°并重复上述过程。如果需要的话，有运营商repea吨的旋转操作高达3倍（共4个夹子）。 </li></ol></li><li>有运营商申请直接反压至切口处用无菌纱布，同时去除大腿活检针，小心不要赶筋膜或皮肤。如果显著感觉到阻力，重新切割，同时将扭转运动到内套管针。 </li><li>在除去从大腿的活检针，具有操作者拉动内套管针背1cm至目视检查外套管的内腔中，如果收集组织的足够量来估计。 <ol><li>有操作者手的活检针至技术员＃1。为了保持无菌技术确保技师＃1只顶住针，而不是运营商。有技术人员＃1手活检针来技师＃3。 </li></ol></li><li>具有技师＃3从活检针的组织。 <ol><li>具有技师＃3使用柱塞（ 图1）和一对精尖镊子以确保所有的肌肉组织从内套管和外套管除去。 </li><li>有技师＃3迅速称重样品，以确认被收集肌肉组织的足够量和放置样品的冰冷解剖块上。 </li></ol></li><li>有技术人员3＃仔细解剖可见结缔组织和脂肪的肌肉样本。已技师＃3，根据下游分析制备的肌肉样本进行存储。例如，立即将样品在离心管和捕捉冷冻在液氮中，安装并冻结了组织学，或发生在离心管与RNA酶抑制剂等。 </li><li>如有必要，重复该步骤与第二灭菌活检针，同时保持无菌操作。 </li></ol> 3，封闭<ol><li>有运营商直接施加压力，活检部位用无菌纱布和冰敷10-15分钟。 </li><li>当止血实现，有运营商使用酒精公关EP垫去除干涸的血迹从切口周围的区域。 </li><li>有技术人员＃1目前局部手术胶粘剂涂抹到运营商的管道。有操作者装配手术粘合剂和施加器的管不接触涂抹器头或管的顶端。 </li><li>有操作者拉，用一只手的切口闭合而施加外科粘合剂的单一层以上的干切口用另一只手的顶部。该粘合剂固化（〜90秒）后，将磁带closuresperpendicular的切口。另外，关闭切口用两个4-0的无菌缝合。 </li><li>有操作者施加直接压力通过放置2月3日至4日"×2"非无菌纱布垫的切口部位，并与自粘合粘接缠绕固定。 </li><li>有运营商提供的主体语言和适当的伤口护理，正常和异常反应的书面说明，并为下面的1〜4天的活动准则（在网上看到苏pplement 2）。 </li></ol>

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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=Brief+intense+interval+exercise+activates+AMPK+and+p38+MAPK+signaling+and+increases+the+expression+of+PGC-1alpha+in+human+skeletal+muscle.">Brief intense interval exercise activates AMPK and p38 MAPK signaling and increases the expression of PGC-1alpha in human skeletal muscle.</a> J Appl Physiol. 106, 929-934 (1985).</li><li>Phillips, S. M., Tipton, K. D., Ferrando, A. A., Wolfe, R. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Resistance+training+reduces+the+acute+exercise-induced+increase+in+muscle+protein+turnover.">Resistance training reduces the acute exercise-induced increase in muscle protein turnover.</a> Am J Physiol. 276, 118-124 (1999).</li><li>Davis, J. M., Murphy, E. A., Carmichael, M. D., Davis, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quercetin+increases+brain+and+muscle+mitochondrial+biogenesis+and+exercise+tolerance.">Quercetin increases brain and muscle mitochondrial biogenesis and exercise tolerance.</a> Am J Physiol Regul Integr Comp Physiol. 296, 1071-1077 (2009).</li><li>Vissing, K., Andersen, J. L., Schjerling, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Are+exercise-induced+genes+induced+by+exercise.">Are exercise-induced genes induced by exercise.</a> FASEB J. 19, 94-96 (2005).</li><li>Lundby, C., 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=Gene+expression+in+human+skeletal+muscle:+alternative+normalization+method+and+effect+of+repeated+biopsies.">Gene expression in human skeletal muscle: alternative normalization method and effect of repeated biopsies.</a> Eur J Appl Physiol. 95, 351-360 (2005).</li><li>Coyle, E. F., Coggan, A. R., Hemmert, M. K., Ivy, J. 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+glycogen+utilization+during+prolonged+strenuous+exercise+when+fed+carbohydrate.">Muscle glycogen utilization during prolonged strenuous exercise when fed carbohydrate.</a> J Appl Physiol. 61, 165-172 (1985).</li></ol>

The authors have no conflicts of interest.

这部影片提供了在亚利桑那州立大学人类表现实验室使用的骨骼肌活检过程的步骤一步的总结。此过程中，小的修改，已被用于在过去的13年，收集约1600肌肉活检。肌肉活检样本中的运动营养基础研究提供了重要的数据，从而导致重要的研究发现。 有需要注意进行改性BERGSTROM骨骼肌活检技术时的许多重要的技术。首先，活检队必须在程序的各个阶段保持严格无菌操作。当定位切口部位[1.4]值得注意的是，股外侧肌，收缩时，比将是在松弛状态下的穿刺过程中更前部和近端是很重要的。如果研究过程需要额外的活检来自同一大腿，初始切口部位应稍存款保险计划一个第三点[1.4]的河谷;每个随后的活组织检查是2-3厘米近端前之一。该计量近似皮褶厚度[1.6]有助于操作者来估计组织的量浅到筋膜，一个重要的判定为步骤2.3.2，2.4.1，2.6，2.6.1和2.11.The操作者必须学会感测在皮下组织和筋膜[2.4.1]之间的电阻变化。当施用局部麻醉剂，这意义将保证肌肉活检的质量通过使操作员能充分地渗入皮下组织，同时最小化的骨骼肌组织直接接触myotoxic 17-19局部麻醉剂[2.4.2]。通过皮下组织切割时，这个电阻变化感会向操作员提供的筋膜[2.6]的深度的指示。操作者应进行心理注意到有多深手术刀是相对于皮肤和切口的相对位置在筋膜[2.6.1]。这将减少的时间由操作者探测筋膜切开所花费的金额和帮助操作者确定所述活检针的窗口是否是完全的肌肉。在外套管（ 图1）的整个窗口必须在肌肉筋膜。以非常稀薄受试者这很容易通过目视确认，然而，这是比较困难的，以确保在受试者具有相当的皮下组织。技师＃1必须学会衡量吸力施加的程度时，注射器被拉开[2.12.1]。低吸入可能表明在枪头 - 活检针结或活检针插入肌肉未焊透弱联合。这两种情况下增加一个重复活检，有必要实现肌肉组织的足够量的可能性。与经验，技术人员＃1也应估计组织根据抗性程度截取的量感到当吸力为being应用。这个信息应该被转发到运营商。类似地，操作员可以评估基于封闭的内套管[2.12.2]当作出切割的"感觉"组织剪切的量。操作人员必须迅速夹肌后技术员＃1用注射器[2.12.2]产生吸力。裁剪的肌肉组织在最大吸点会改善组织的产量。与活检针的单次通过的多个剪辑的方法有助于确保在该研究中获得的组织足量和通过减小活检针插入肌肉通过筋膜切开的次数减少了受试者的不适[ 2.12.4。运营商应该从切口缓慢，扭转运动，而使用他们腾出手来申请用无菌纱布[2.13]反压取出活检针。这将有助于防止获取活检针的窗口中的筋膜和皮肤"挂了"cisions。性的感觉，同时抽取针可能表明通过肌肉的不完全切割。如果一个不完整的切割之嫌，运营商应停止拆除活检针通过打开内套管针4-5毫米重新切割剩余的纤维，并应用轻微的扭转运动作为内套管针被关闭。最后，取出活检针后，冷敷，以切口部位10-15分钟，有些科目可能需要更长的时间来达到止血[3.1]。止血已经达到后，仔细清洗切口周围用酒精消毒焊盘的面积，并且让皮肤干燥施加的外科粘合剂[3.2-3.4]之前。这些技术将最大限度地利用外科粘合剂关闭皮肤切口和粘性绷带的有效性粘到皮肤的益处。手术粘合拥有带封锁和缝线几个优点。受试者可以大力立即多项式运动后数小时杜热而不从切口出血。该手术的粘合剂提供了对潜在的感染一种物理性屏障，不要求去除，并最大限度地减少疤痕。 肌肉活检程序是微创的，比较安全，并且可在生物医学研究中的设置作为门诊病人程序进行。大多数受试者报告他们进行生活正常的日常活动能力变化不大。疼痛和从程序肿胀是在5至7天完全解决。很少有不良反应的报道，从参与在亚利桑那州立大学人类表现实验室进行的研究课题（约2超过了1600，无论是局部皮肤感染）。这个发病率是相似的其他已发表的报告。伯格斯坦报道3非常低的肌肉血肿的发生率，这自然再吸收，只有1例出了5000多活检需要手术干预，以阻止动脉出血。亨ssey 等人 ，采用改良伯格斯坦技术的抽吸，报道称，2活检出83导致因疼痛和不适主体的日常活动的干扰。其中一个事件是由于一个瘀斑，以及两个事件7天内6解决。 Tarnopolsky 等 。报告一个非常低的并发症发生率与吸修改伯格斯坦技术（22并发症出13,914活检在成人和儿童7）。并发症有局部皮肤感染（8例），动脉出血（2例），瘀斑/血肿（2例），疼痛持续&gt; 3天（5例），小面积局部麻木远端活检（5例） 7。 修改后的伯格斯坦肌肉活检技术提升的研究过程中，当施加最小的时间限制，所收集的数据的质量。该过程可以迅速完成，届时切口部位是提前准备的时间，一个PAS3-4剪辑Š通常会产生组织100至200毫克，而多点活检可在单个会话期间取自一个主题。用这种方法收集的肌肉样品，可用于对结果的措施，包括纤维打字，肌肉损伤，燃料基材的商店，线粒体生物合成和呼吸，酶活性，在代谢的变化，蛋白质的合成，基因表达的炎症和氧化应激的广泛阵列，和无数人。 从股外侧肌得到肌肉活检样品提供优于其他候选的肌肉（如肱二头肌，三角肌，腓肠肌，胫前肌，肱三头肌，和比目鱼肌）。股外侧是最常用的采样肌肉，以及广泛的规范性组织学，生物化学的数量，并为研究人员和临床分子生物学的数据存在比较他们的调查结果和意见。其次，股的区域外侧，其中INCIsions是由比较大并且不具有大血管或神经重叠的区域。因此，一个给定的测试会话期间在同一肌肉多个切口应，每2-3厘米分开，以防止在基因表达的24外伤引起的变化。取得从通过单独的切口相同的肌肉多次活检没有明显改变的基因响应于锻炼24,25的应力的转录响应。但是，当第二次活检是通过相同的切口采取几个小时以后24的某些基因的表达可受到影响。最后，受试者能够在从股外侧肌中获得的肌肉活检从事2-3小时的运动，很少不适。很多科目后活检不适比较温和的挫伤。这使得前向运动后的样品几乎没有受到负担3,8,11,26获得。 在摘要中，一种方法为提取使用改性BERGSTROM技术人骨骼肌活检样品进行了说明。这种肌肉活检过程是比较安全的，并提供了充足的骨骼肌组织多个下游的细胞和分子检测的研究。该过程可以迅速完成，从而使研究者能够获得前，中，后的运动（或其他干预）活检样品。

经皮或"半开放"活检技术被用来获得从人患者和研究受试者的骨骼肌组织标本。假肥大型（1806年至1875年），相信作为第一个构建针头套管针经皮活检1以获取活体骨骼肌。在20世纪60年代，伯格斯坦推出了经皮活检针类似由杜氏2-4描述。二十年后，Evans 等人的 5通过经由BERGSTROM针的切割套管针施加抽吸改性的技术。这种修饰可以增强组织3的产率，以5倍6,7和被用在临床和生物医学研究的设置。这种技术已经并将继续培养肌病的诊断和我们的骨骼肌的结构和功能的理解。 经皮肌活检技术是straightforward。如果做得正确，严格遵守无菌技术操作，相关的风险是最小的。肌肉活检程序被视为一种研究工具，用于研究项目。一些学术机构允许经过培训的教师与研究人员博士，与医生的直接监督，以获得肌肉活检等人要求委员会认证的医师来执行的技术。在过去的13年8月15日在亚利桑那州立大学活检团队已经成功完成超过1,600肌肉活检。该视频的目的是描述了改性BERGSTROM针经皮肌活检技术5获得骨骼肌组织样品从人类受试者的股外侧。

<table><tbody><tr><td>Bergstr&amp;ouml;m biopsy needle, 5 x 100 mm</td><td>Stille Surgical Inc.</td><td>119-29187-50</td><td>1 ea</td></tr><tr><td>Durasorb blue pad</td><td>Fisher</td><td>22-031-340</td><td>case of 300</td></tr><tr><td>Prep razor disposable</td><td>Moore Medical</td><td>89760</td><td>1 ea</td></tr><tr><td>Shave cream</td><td>Moore Medical</td><td>92869</td><td>1 can</td></tr><tr><td>Alcohol swab</td><td>Moore Medical</td><td>98721</td><td>box/200</td></tr><tr><td>Povidone-iodine swabsticks (3 pack)</td><td>Moore Medical</td><td>90691</td><td>pack of 50</td></tr><tr><td>Face masks</td><td>Fisher</td><td>19-039-690</td><td>case of 50</td></tr><tr><td>Sterile gloves; sz 6.5</td><td>Moore Medical</td><td>68128</td><td>box of 50 pairs</td></tr><tr><td>Sterile gloves; sz 7.0</td><td>Moore Medical</td><td>68129</td><td>box of 50 pairs</td></tr><tr><td>Sterile gloves; sz 8.5</td><td>Moore Medical</td><td>68132</td><td>box of 50 pairs</td></tr><tr><td>Fenestrated towel drape</td><td>Moore Medical</td><td>92712</td><td>case of 50</td></tr><tr><td>Lidocaine (1% HCl, w/o Epi)</td><td>Dealmed</td><td>427902</td><td>30 cc vial</td></tr><tr><td>Ethyl chloride spray</td><td>Dealmed</td><td>386020</td><td>3.5 fl. Oz. bottle</td></tr><tr><td>5 ml syringe w/ 21 G needle</td><td>Fisher</td><td>14-827-48</td><td>case of 100</td></tr><tr><td>25 G 5/8&quot; needle</td><td>Fisher</td><td>14-826AA</td><td>case of 100</td></tr><tr><td>25 G 1 1/2&quot; needle</td><td>Fisher</td><td>14-826-49</td><td>case of 100</td></tr><tr><td>Single-use scalpels w/ #11 blade</td><td>Fisher</td><td>0-8927-5B</td><td>box of 20</td></tr><tr><td>60 ml Syringe</td><td>Fisher</td><td>13-689-8</td><td>case of 40</td></tr><tr><td>~30 inch extension tubing</td><td>Fisher</td><td>14-169-7A</td><td>roll of 50 ft</td></tr><tr><td>Metal stopcock</td><td>Fisher</td><td>01-290-38</td><td>each</td></tr><tr><td>Polypropylene tubing connector w/ tapered ends, 3.2 to 5.5 mm</td><td>Fisher</td><td>15-315-28A</td><td>case of 100</td></tr><tr><td>Yellow pipette tips (1-200 &amp;micro;l)</td><td>Fisher</td><td>02-681-2</td><td>cut off ~15-18 mm of the tip</td></tr><tr><td>Topical surgical adhesive</td><td>Medline</td><td>MSC091076</td><td>case of 36 tubes</td></tr><tr><td>Topical surgical adhesive</td><td>Medline</td><td>MSC091076Z</td><td>box of 6 tubes</td></tr><tr><td>Butterfly stitches (adhesive strips) 1/2&quot; x 4&quot; (6 per pack)</td><td>Moore Medical</td><td>93050</td><td>box/300</td></tr><tr><td>Bandadge, Strip, 3/4 x 3 in.</td><td>Fisher</td><td>19-027-202</td><td>pack of 100</td></tr><tr><td>Adhesive bandage (5 m rolls)</td><td>Fisher</td><td>19-027-768</td><td>case of 36</td></tr><tr><td>1&quot; paper surgical tape</td><td>Fisher</td><td>19-027-761</td><td>case of 12</td></tr><tr><td>Sutures (4-0 reverse cutting, FS2)</td><td>Moore Medical</td><td>82926</td><td>box of 36</td></tr><tr><td>4 x 4 Sterile gauze pads</td><td>Moore Medical</td><td>66949</td><td>pkg/50</td></tr><tr><td>2 x 2 Non-sterile gauze pads</td><td>Moore Medical</td><td>37336</td><td>pkg/200</td></tr><tr><td>3 x 4 Non-adherent pads</td><td>Moore Medical</td><td>74389</td><td>bx/100</td></tr><tr><td>Autoclave bags 3.5 x 9</td><td>Moore Medical</td><td>40390</td><td>box of 200</td></tr><tr><td>Autoclave bags 5.25 x 10</td><td>Moore Medical</td><td>40391</td><td>box of 200</td></tr><tr><td>Autoclave indicator tape</td><td>Moore Medical</td><td>31608</td><td>box of 250</td></tr><tr><td>Lab coats&amp;mdash;small</td><td>Fisher</td><td>23-900-520A&amp;nbsp;</td><td>10/pk</td></tr><tr><td>Lab coats&amp;mdash;medium</td><td>Fisher</td><td>23-900-520B&amp;nbsp;</td><td>10/pk</td></tr><tr><td>Lab coats&amp;mdash;large</td><td>Fisher</td><td>23-900-520C&amp;nbsp;</td><td>10/pk</td></tr><tr><td>Germicidal Disposable Cloth Wipes</td><td>Moore Medical</td><td>62879</td><td>Can/160</td></tr></tbody></table>

human skeletal muscle biopsy procedures using the modified bergstr&#246;m technique

经皮活检技术使研究人员和临床医生收集的骨骼肌肉组织样本。该技术是安全的和高度有效的。这部影片描述了使用修改后的伯格斯坦针，以获得人类受试者的股外侧肌肉组织样本经皮活检技术。该BERGSTROM针包括一个外套管具有在前端侧的小开口（"窗口"）和一个内套管针带切削刃的前端。在局部麻醉和无菌条件下，将针通过皮肤，皮下组织和筋膜切开前进到骨骼肌。接着，抽吸施加到内套管针，所述外套管针被拉回，骨骼肌组织被吸入到所述外套管被抽吸的窗口，并且所述内套管针迅速闭合，从而切断或削波的骨骼肌组织样品。针旋转90°60，另有裁切。这个过程可以重复3次以上。此多刀切割技术通常产生100-200毫克或更多的健康受试者的样品与可以做立即之前，期间和运动或其他介入的回合之后。随着切口部位的后活检敷料，受试者通常恢复日常活动的时候了，并能充分参与身体活动中的48-72小时。受试者应避免重阻力锻炼48小时通过切口在筋膜，减少肌肉症的风险。

如上文所描述的肌肉活检程序允许研究者快速一致收集骨骼肌组织样品。典型的产率在健康，运动受试者是200毫克或更多的单次通过以3-4的剪辑。该过程需要15-20分钟，其中大部分是花在准备在切口。在运动为基础的研究，肌肉样品往往采取前后运动，以恢复过程中收集的一个或两个样本。利用这一设计，在运动后的毛发（对侧）大腿被限幅和切口位点是预先标有一个细点永久性标记，而运动前大腿正被用于的方法制备。这样做允许在20或长时间运动的一个回合后 ​​应采取的8-10分钟的肌肉活检。另外，调查工作的时间较短时（ 如 30分钟或更少）的切口（S），可事先向运动，盖上消毒敷料，并拧紧编带手术胶带，由此，允许所述活检样本被快速地完成一次运动后拍摄（10秒21到4分钟，22）。该过程的相对快速性使得研究人员才捕捉到细胞和分子事件，期间和运动的回合后，并测试营养干预的互动效应。 亚利桑那州立大学人类表现实验室发表了利用来自肌肉组织活检样本数据的几种运动营养学基础试卷。来自亚利桑那州立大学的研究小组取得了重要的科学发现包括: IL-1β的基因的表达，IL-6，IL-8和TNF-α是在3小时的运转后，由有经验的耐力运动员获得骨骼肌组织增加。碳水化合物的摄入（60克/小时）减毒的IL-6和IL-8的表达水平，尽管具有对骨骼肌糖原耗竭8（ 图2）没有任何影响。 ="jove_content"&gt;有2小时的密集阻力训练回合增加肌肉组织的细胞因子mRNA的IL-1β，IL-6，IL-8和TNF-α的30名经验丰富的选手（即相同的细胞因子的升高下运行） 。在肌肉组织中表达的增幅很大，但没有相应增加血浆细胞因子水平。碳水化合物的摄入没有影响变化的肌肉组织的细胞因子mRNA表达9的图形。 碳水化合物摄取由减毒血浆IL-6 15训练有素的骑自行车的人与安慰剂相比，却不得不IL-8，TNF-α的基因表达的自行车11的2.5小时的肌肉组织糖原耗竭或IL-6没有影响。 槲皮素（1,000 mg /天，连续3周）摄取40训练有素的骑自行车的人不改变运动引起的增加的肌肉组织中的NF-κB，COX-2，IL-1β，IL-6，IL-8，或TNF-α的mRNA的与安慰剂组15式相比。 jove_content"&gt;相较于哪些被发现在小鼠23 1000毫克/天，槲皮素2周对线粒体生物合成的26人未受过训练的科目13个标记不大，但不显着，效果。 <img alt="图2" fo:content-width="2in" src="/files/ftp_upload/51812/51812fig2highres.jpg" width="300" /> 从人的骨骼肌肉组织活检得到图2的数据。在一项随机，安慰剂对照，交叉研究的男性的耐力运动员组（n = 16），跑跑步机3小时，70％VO2max最大摄氧量和消耗6％的碳水化合物饮料或安慰剂饮料以1升/小时8的速度。骨骼肌组织切片检查被前后运动，立即置于冰 - 冷夹层块上，并可见结缔组织和脂肪小心地除去。活检样品然后放置在一个离心管，在液氮中冷冻，并保存于-8<html> 0℃，直到分析（一）碳水化合物摄入不影响骨骼肌糖原耗竭（P = 0.246，交互作用）。但是，碳水化合物摄取倾向于分别衰减的增加（B）的骨骼肌组织白细胞介素（IL）-6 mRNA水平，和（C）的骨骼肌组织中IL-8的mRNA水平（P = 0.071和P = 0.063，交互作用， ）8。 <table border="0" cellpadding="0" cellspacing="0" fo:keep-together.within-page="always" width="511"><tbody><tr height="42"><td height="42" style="height:42px;width:355px;"> 项目 </td><td style="width:156px;"> 每活检数量 </td></tr><tr height="21"><td height="21" style="height:21px;">手套，非无菌</td><td> 3 </td></tr><tr height="21"><td height="21" style="height:21px;"> Durasorb蓝色垫</td><td> 2 </td></tr><tr height="21">;"&gt;口罩</td><td> 1 </td></tr><tr height="21"><td height="21" style="height:21px;">准备剃须刀，一次性</td><td> 1 </td></tr><tr height="21"><td height="21" style="height:21px;">剃须霜</td><td>部分即可</td></tr><tr height="21"><td height="21" style="height:21px;">记号笔/不可磨灭的标记</td><td>可重复使用</td></tr><tr height="21"><td height="21" style="height:21px;">聚维酮碘Swabsticks（3包） </td><td> 1 </td></tr><tr height="21"><td height="21" style="height:21px;">无菌手术手套（大小而定） </td><td> 1 </td></tr><tr height="21"><td height="21" style="height:21px;">有孔垂坠巾</td><td> 1 </td></tr><tr height="21"><td height="21" style="height:21px;">氯乙烷喷雾</td><td> 0.1 </td></tr><tr height="21"><td height="21" style="height:21px;">利多卡因（1％W / O型EPI） </td><td> 5毫升</td></tr><tr height="21"><td height="21" sty勒="高度:21px;"&gt; 5毫升注射器W / 21G针头</td><td> 1 </td></tr><tr height="21"><td height="21" style="height:21px;">地下25 5/8"针</td><td> 1 </td></tr><tr height="21"><td height="21" style="height:21px;"> 25 G 1半"针</td><td> 1 </td></tr><tr height="21"><td height="21" style="height:21px;">一次性使用手术刀W /＃11刀</td><td> 1 </td></tr><tr height="21"><td height="21" style="height:21px;">无菌的4×4的纱布垫</td><td> 4 </td></tr><tr height="21"><td height="21" style="height:21px;">包装的消毒伯格斯坦活检针</td><td> 1 </td></tr><tr height="21"><td height="21" style="height:21px;"> 60毫升注射器</td><td> 1 </td></tr><tr height="21"><td height="21" style="height:21px;">延长管</td><td> 〜75公分</td></tr><tr height="21"><td height="21" style="height:21px;">金属活塞</td><td> 1 </td></tr><tr height="21"><td heighT ="21"的风格="高度:21px;"&gt;塑料连接器</td><td> 1 </td></tr><tr height="21"><td height="21" style="height:21px;">切割黄色（1-200μL）枪头</td><td> 1 </td></tr><tr height="21"><td height="21" style="height:21px;">酒精棉签</td><td> 5-10 </td></tr><tr height="21"><td height="21" style="height:21px;"> Octylseal，液体缝线</td><td> 1 </td></tr><tr height="21"><td height="21" style="height:21px;">免缝条片半"×4" </td><td> 1 </td></tr><tr height="21"><td height="21" style="height:21px;">绷带</td><td> 1 </td></tr><tr height="21"><td height="21" style="height:21px;">非无菌的2×2（或4×4）纱布垫</td><td> 2（或1）的</td></tr><tr height="21"><td height="21" style="height:21px;">粘性绷带（5米卷） </td><td> 0.25 </td></tr><tr height="21"><td height="21" style="height:21px;">纸带固定胶粘绷带</td><td> 〜30公分</td></tr&gt; </tbody></table> 表1。肌肉活检材料清单。

Watch this Scientific Journal Video about Human Skeletal Muscle Biopsy Procedures Using the Modified BergstrÃƒÂ¶m Technique at JoVE.com

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

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

人骨骼肌活检手术使用修改伯格斯坦技术

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

human-skeletal-muscle-biopsy-procedures-using-modified-bergstrm

Nanyang Technological University

Research

JoVE Journal

Medicine

39.8K 次观看.  Appalacian State University. The purpose of this video is to present the modified Bergström skeletal muscle biopsy technique on human subjects. 

视频: Human Skeletal Muscle Biopsy Procedures Using the Modified Bergström Technique

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.

从人类骨骼肌成肌原细胞和成纤维细胞的分离和定量免疫细胞化学表征

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

科研

发育生物学

Myo-mechanical Analysis of Isolated Skeletal Muscle

To assess the in vivo effects of therapeutic interventions for the treatment of muscle disease 1,2,3, quantitative methods are needed that measure force generation and fatigability in treated muscle. We describe a detailed approach to evaluating myo-mechanical properties in freshly explanted hindlimb muscle from the mouse. We describe the atraumatic harvest of mouse extensor digitorum longus muscle, mounting the muscle in a muscle strip myograph (Model 820MS; Danish Myo Technology), and the measurement of maximal twitch and tetanic tension, contraction time, and half-relaxation time, using a square pulse stimulator (Model S48; Grass Technologies). Using these measurements, we demonstrate the calculation of specific twitch and tetanic tension normalized to muscle cross-sectional area, the twitch-to-tetanic tension ratio, the force-frequency relationship curve and the low frequency fatigue curve 4. This analysis provides a method for quantitative comparison between therapeutic interventions in mouse models of muscle disease 1,2,3,5, as well as comparison of the effects of genetic modification on muscle function 6,7,8,9.

To assess the in vivo effects of therapeutic interventions for muscle disease, methods are needed to quantitate force generation and fatigability in treated muscle. We detail an approach to evaluating myo-mechanical properties in explanted mouse hindlimb muscle. This analysis provides a robust approach to quantitating the effects of genetic modification on muscle function, as well as comparison of therapies in mouse models of muscle disease.

离体骨骼肌肌的力学分析

To assess the in vivo effects of therapeutic interventions for the treatment of muscle disease 1,2,3, quantitative methods are needed that measure force ...

医学

X-ray Diffraction of Intact Murine Skeletal Muscle as a Tool for Studying the Structural Basis of Muscle Disease

Transgenic mouse models have been important tools for studying the relationship of genotype to phenotype for human diseases including those of skeletal muscle. Mouse skeletal muscle has been shown to produce high quality X-ray diffraction patterns on third generation synchrotron beamlines providing an opportunity to link changes at the level of the genotype to functional phenotypes in health and disease by determining the structural consequences of genetic changes. We present detailed protocols for preparation of specimens, collecting the X-ray patterns and extracting relevant structural parameters from the X-ray patterns that may help guide experimenters wishing to perform such experiments for themselves.

We present detailed protocols for performing small-angle X-ray diffraction experiments using intact mouse skeletal muscles. With the wide availability of transgenic mouse models for human diseases, this experimental platform can form a useful test bed for elucidating the structural basis of genetic muscle diseases

完整毛里丝骨骼肌的X射线衍射作为研究肌肉疾病结构基础的工具

Transgenic mouse models have been important tools for studying the relationship of genotype to phenotype for human diseases including those of skeletal ...

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.

从骨骼肌活检程序分离、培养、表征和分化人类肌肉祖细胞

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

Performing Human Skeletal Muscle Xenografts in Immunodeficient Mice

Treatment effects observed in animal studies often fail to be recapitulated in clinical trials. While this problem is multifaceted, one reason for this failure is the use of inadequate laboratory models. It is challenging to model complex human diseases in traditional laboratory organisms, but this issue can be circumvented through the study of human xenografts. The surgical method we describe here allows for the creation of human skeletal muscle xenografts, which can be used to model muscle disease and to carry out preclinical therapeutic testing. Under an Institutional Review Board (IRB)-approved protocol, skeletal muscle specimens are acquired from patients and then transplanted into NOD-Rag1null IL2r&gamma;null (NRG) host mice. These mice are ideal hosts for transplantation studies due to their inability to make mature lymphocytes and are thus unable to develop cell-mediated and humoral adaptive immune responses. Host mice are anaesthetized with isoflurane, and the mouse tibialis anterior and extensor digitorum longus muscles are removed. A piece of human muscle is then placed in the empty tibial compartment and sutured to the proximal and distal tendons of the peroneus longus muscle. The xenografted muscle is spontaneously vascularized and innervated by the mouse host, resulting in robustly regenerated human muscle that can serve as a model for preclinical studies.

Complex human diseases can be challenging to model in traditional laboratory model systems. Here, we describe a surgical approach to model human muscle disease through the transplantation of human skeletal muscle biopsies into immunodeficient mice.

在免疫缺陷小鼠中表演人类骨骼肌异种移植物

Treatment effects observed in animal studies often fail to be recapitulated in clinical trials. While this problem is multifaceted, one reason for this ...

Quantification of Subcellular Glycogen Distribution in Skeletal Muscle Fibers using Transmission Electron Microscopy

With the use of transmission electron microscopy, high-resolution images of fixed samples containing individual muscle fibers can be obtained. This enables quantifications of ultrastructural aspects such as volume fractions, surface area to volume ratios, morphometry, and physical contact sites of different subcellular structures. In the 1970s, a protocol for enhanced staining of glycogen in cells was developed and paved the way for a string of studies on the subcellular localization of glycogen and glycogen particle size using transmission electron microscopy. While most analyses interpret glycogen as if it is homogeneously distributed within the muscle fibers, providing only a single value (e.g., an average concentration), transmission electron microscopy has revealed that glycogen is stored as discrete glycogen particles located in distinct subcellular compartments. Here, the step-by-step protocol from tissue collection to the quantitative determination of the volume fraction and particle diameter of glycogen in the distinct subcellular compartments of individual skeletal muscle fibers is described. Considerations on how to 1) collect and stain tissue specimens, 2) perform image analyses and data handling, 3) evaluate the precision of estimates, 4) discriminate between muscle fiber types, and 5) methodological pitfalls and limitations are included.

A modified post-fixation procedure increases the contrast of glycogen particles in tissue. This paper provides a step-by-step protocol describing how to handle the tissue, conduct the imaging, and use stereological methods to obtain unbiased and quantitative data on fiber type-specific subcellular glycogen distribution in skeletal muscle.

使用透射电子显微镜定量骨骼肌纤维中的亚细胞糖原分布

With the use of transmission electron microscopy, high-resolution images of fixed samples containing individual muscle fibers can be obtained. This ...

生物化学

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.

线粒体的制备及透气性评估，从骨骼肌组织通过穿刺活检获得的分离

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

MyDoBID- 一种有效识别肌肉纤维类型的新技术

Fiber Type Identification of Human Skeletal Muscle

The technique described here can be used to identify specific myosin heavy chain (MHC) isoforms in segments of individual muscle fibers using dot blotting, hereafter referred to as Myosin heavy chain detection by Dot Blotting for IDentification of muscle fiber type (MyDoBID). This protocol describes the process of freeze-drying human skeletal muscle and isolating segments of single muscle fibers. Using MyDoBID, type I and II fibers are classified with MHCI- and IIa-specific antibodies, respectively. Classified fibers are then combined into fiber type-specific samples for each biopsy.
The total protein in each sample is determined by Sodium Dodecyl-Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) and UV-activated gel technology. The fiber type of samples is validated using western blotting. The importance of performing protein loading normalization to enhance target protein detection across multiple western blots is also described. The benefits of consolidating classified fibers into fiber type-specific samples compared to single-fiber western blots, include sample versatility, increased sample throughput, shorter time investment, and cost-saving measures, all while retaining valuable fiber type-specific information that is frequently overlooked using homogenized muscle samples. The purpose of the protocol is to achieve accurate and efficient identification of type I and type II fibers isolated from freeze-dried human skeletal muscle samples.
These individual fibers are subsequently combined to create type I and type II fiber type-specific samples. Furthermore, the protocol is extended to include the identification of type IIx fibers, using Actin as a marker for fibers that were negative for MHCI and MHCIIa, which are confirmed as IIx fibers by western blotting. Each fiber type-specific sample is then used to quantify the expression of various target proteins using western blotting techniques.

作者聚焦:使用 MyDoBID 技术破译骨骼肌纤维类型的奥秘 

This protocol demonstrates single-fiber isolation from freeze-dried human skeletal muscle and fiber-type classification according to Myosin heavy chain (MHC) isoform using the dot blotting technique. Identified MHC I and II fiber samples can then be further analyzed for fiber type-specific differences in protein expression using western blotting.

人骨骼肌活检手术使用修改伯格斯坦技术

摘要

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此视频中的章节

从人类骨骼肌成肌原细胞和成纤维细胞的分离和定量免疫细胞化学表征

离体骨骼肌肌的力学分析

完整毛里丝骨骼肌的X射线衍射作为研究肌肉疾病结构基础的工具

从骨骼肌活检程序分离、培养、表征和分化人类肌肉祖细胞

在免疫缺陷小鼠中表演人类骨骼肌异种移植物

使用透射电子显微镜定量骨骼肌纤维中的亚细胞糖原分布

线粒体的制备及透气性评估，从骨骼肌组织通过穿刺活检获得的分离

人体骨骼肌的纤维类型鉴定

人体骨骼肌的纤维类型鉴定

人体骨骼肌的纤维类型鉴定